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Journal of Dietary Supplements
ISSN: 1939-0211 (Print) 1939-022X (Online) Journal homepage: https://www.tandfonline.com/loi/ijds20
Pharmacological evaluation of Ashwagandha
highlighting its healthcare claims, safety, and
toxicity aspects
Deepa S. Mandlik (Ingawale) & Ajay G. Namdeo
To cite this article: Deepa S. Mandlik (Ingawale) & Ajay G. Namdeo (2020): Pharmacological
evaluation of Ashwagandha highlighting its healthcare claims, safety, and toxicity aspects, Journal
of Dietary Supplements, DOI: 10.1080/19390211.2020.1741484
To link to this article: https://doi.org/10.1080/19390211.2020.1741484
Published online: 03 Apr 2020.
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REVIEW
Pharmacological evaluation of Ashwagandha highlighting
its healthcare claims, safety, and toxicity aspects
Deepa S. Mandlik (Ingawale), PhD and Ajay G. Namdeo, PhD
Department of Pharmacology, Bharati Vidyapeeth Deemed to be University, Poona College of
Pharmacy, Erandwane, Pune, India
ABSTRACT
Withania somnifera, commonly known as “Ashwagandha” or “Indian
ginseng” is an essential therapeutic plant of Indian subcontinent
regions. It is regularly used, alone or in combination with other
plants for the treatment of various illnesses in Indian Systems of
Medicine over the period of 3,000 years. Ashwagandha (W. somnifera)
belongs to the genus Withania and family Solanaceae. It comprises
a broad spectrum of phytochemicals having wide range of biological
effects. W. somnifera has demonstrated various biological actions
such as anti-cancer, anti-inflammatory, anti-diabetic, anti-microbial,
anti-arthritic, anti-stress/adaptogenic, neuro-protective, cardio-
protective, hepato-protective, immunomodulatory properties.
Furthermore, W. somnifera has revealed the capability to decrease
reactive oxygen species and inflammation, modulation of mitochon-
drial function, apoptosis regulation and improve endothelial function.
Withaferin-A is an important phytoconstituents of W. somnifera
belonging to the category of withanolides been used in the
traditional system of medicine for the treatment of various disorders.
In this review, we have summarized the active phytoconstituents,
pharmacologic activities (preclinical and clinical), mechanisms of
action, potential beneficial applications, marketed formulations and
safety and toxicity profile of W. somnifera.
KEYWORDS
adaptogenic; anti-
Alzheimer; anti-arthritic;
anti-diabetic; anti-cancer;
anti-hypoxic; anti-
inflammatory; anti-ischemic;
anti-microbial; anti-
Parkinson; anti-stress;
aphrodisiac; cardio-
protective; clinical
evaluation; hepatoprotec-
tion; immunomodulatory;
neuro-protective; safety and
toxicity; Withania somnifera
CONTACT Deepa S. Mandlik (Ingawale) deepa_ingawale@yahoo.com Department of Pharmacology, Bharati
Vidyapeeth Deemed to be University, Poona College of Pharmacy, Erandwane, Pune- 411038, India.
ß 2020 Taylor & Francis Group, LLC
JOURNAL OF DIETARY SUPPLEMENTS
https://doi.org/10.1080/19390211.2020.1741484

Introduction
Withania somnifera (W. somnifera) is a woody, evergreen shrub of 0.5 to 2.0 m
in height usually identified as “Winter cherry” or “Indian Ginseng” in English,
“Ashwagandha” in Sanskrit, “Asgandh” in Hindi and “Asgand” in Urdu (Ziauddin et al.
1996; Dhuley 1998). The plant belongs to the family Solanaceae and cultivated in hot
and dry parts of tropical and subtropical zones of world. It grows from Canary Islands,
South Africa, Middle East, Sri Lanka, China, India to warmer parts of Europe and Australia
(Purdie et al. 1982; Hepper 1991). The W. somnifera whole plant or its different parts are
commonly used in Ayurvedic and Unani medicine systems of India for its medicinal and
therapeutic usage for more than 5,000years. It is also reported as an official drug in Indian
Pharmacopoeia-1985 (Singh et al. 2011; Uddin et al. 2012) (Figure 1).
It is considered as an important herbal Rasayana and known as “Sattvic Kapha
Rasayana.” Rasayana is herbal or metallic preparation that is used for various
pharmacologic properties such as/in aphrodisiac, adaptogenic, diuretic, anti-helminthic,
astringent, tonic, narcotic, immuno-stimulation, anti-inflammation, anti-stress, anti-
ulcer, rejuvenative, rheumatism, goiter, boils, pimple, piles, flatulent colic, oligospermia,
health promoter, leucoderma, constipation, insomnia, nervous breakdown, snake venom,
and scorpion stings (Agarwal et al. 1999; Machiah et al. 2006; Machiah and Gowda
2006). Furthermore, W. somnifera is used for the clearing of white spots from the
cornea and also used in anxiety, hysteria, syncope, and loss of memory. Ashwagandha
is used for extreme weight loss in children. On the other hand, along with milk, it acts
as tonic for children (Basu 1935; Bhandari 1970; Sharma et al. 1985; Misra 2004; Dar
2 D. S. MANDLIK (INGAWALE) AND A. G. NAMDEO

et al. 2015). Due to its distinct stress-busting qualities, the plant species achieves the
name “somnifera,” which means “sleep-inducer” (Ven Murthy et al. 2010; Seenivasagam
et al. 2011).
The plant is known as Ashwagandha because the plant roots display the distinctive
smell of a wet horse (“ashwa” means horse and “gandha” means smell). Also, it is
known as Indian ginseng due to its pharmacologic and traditional uses are like to that
of Korean ginseng tea (Dar et al. 2015). The pharmacologic activity of plant remains for
less than two years due to the decomposition of its phytoconstituents. Because of this
problem, during the period of January-March, the fresh roots are harvested every year
and shade drying is done for getting better yield and medicinal results. The flowers pos-
sesses diuretic, astringent, depurative, and aphrodisiac properties whereas, leaves are
used for the treatment of fever. The seeds have medicinal properties such as improving
sperm count and testicular growth, anti-helminthic, removing white spots from cornea.
However, fruits are commonly used against various types of skin ulcers, skin diseases
and carbuncles (Chopra et al. 2004; Kaur et al. 2004; Singh et al. 2011). The different
in-vivo and in-vitro pharmacological activities are summarized in Tables 1 and 2.
Bioactive constituents of Withania somnifera
Withanolides is a group of steroidal lactones responsible for the pharmacological
activity of roots of W. somnifera (Figure 2) (Budhiraja and Sudhir 1987). Laboratory
investigations has concluded that over 35 phytoconstituents are present in the roots
of W. somnifera (Rastogi and Mehrotra 1998). Phytochemical analysis has discovered
the occurrence of diverse chemical constituents in different parts of W. somnifera. Up
till now, more than 40 withanolides, 12 alkaloids, and rare sitoindosides have been pre-
sent in the plant (Mirjalili et al. 2009) (Table 3). The most remarkable ingredients are
tropine alkaloids such as Convolamine, Convoline, Convolidine, Convolvine, confoline,
convosine, etc. (Prasad et al. 1974; Lounasmaa 1988; Singh and Bhandari 2000). The
fresh plant of W. somnifera contains Fatty acids, Fatty alcohols, Volatile oils, Myristic
acid, Palmitic acid, Linoleic acid, and Hextriacontane. The roots of W. somnifera con-
tains Reducing sugar, Starch, Glycosides, and Withaniol acid. It also contains eight bases
such as Withanine, Withananine, Withananinine, Pseudowithanine, Withasomnine,
Figure 1. Withania somnifera plant.
JOURNAL OF DIETARY SUPPLEMENTS 3

Table 1. In-vivo pharmacological activities of W. somnifera.
Sr. No. Nature of extract Diseased model Mechanism of action References
1 Root extract 500 mg/ml
and 1000 mg/kg
(rectal route)
TNBS-induced
inflammatory bowel
disease in rats
Muco-restorative and anti-
inflammatory activity,
resolved neutrophil
infiltration, edema,
and necrosis
Pawar et al. (2011)
2 Root extract 500 and
1000 mg/kg
(oral route)
Mouse model of lupus Inhibited proteinuria,
nephritis, TNF-a, NO,
and ROS
Minhas et al. (2011, 2012)
3 Aqueous extract of root
powder 600 and
800 mg/kg (oral route)
Collagen-induced
arthritis in rats
Attenuated cartilage
degradation, improved
the functional recovery
of motor activity, and
radiological score
Gupta and Singh (2014)
4 Aqueous fraction of root
extract 25, 50, 100,
and 200 mg/kg
(oral route)
Mouse model of
chronic stress
Reduced in T-cell
population and up-
regulated
Th1 Cytokines
Khan et al. (2006)
5 Root extract 20 mg/kg
(oral route)
Immobilization stress
in albino rats
Markedly rescued the
number of
degenerating cells in
CA2 and CA3 subareas
of rat hippocampus
6 Root extract 100 mg/kg
(oral route)
MPTP induced toxicity
in mice
Normalized catecholamine
content, reduced
oxidant stress and
functional activity
Sankar et al. (2007) and
Rajasankar et al. (2009)
7 Root powder 100 and
Rotenone-induced
impairment in mice
Antioxidant and anti-
inflammatory actions,
corrected
mitochondrial
dysfunctions,
normalized NT function
Manjunath and
Muralidhara (2013)
8 Root extract 1 g/kg
(oral route)
Alzheimer’s
disease models
Reversed behavioral
deficits, pathological
clues as well as Ab by
up-regulating
lipoprotein receptor
in liver
Sehgal et al. (2012)
9 Root extract 50, 100, 150,
200, and 250 mg/kg
(oral route)
Hypoxia pathway in
hippocampal cells
Enhanced memory and
attenuated
hippocampal
neurodegeneration by
repleting
glutathione levels
Baitharu et al.
(2013, 2014)
10 Whole extract 30, 60, and
Myocardial infarction
in Rats
Cardiotropic and
cardioprotective
activity
Ojha and Arya (2009),
Prince et al. (2008),
and Reuland
et al. (2013)
11 Whole extract 50 mg/kg
(oral route)
Myocardial infarction
in Rats
Anti-apoptotic/pro-
apoptotic effects and
reduced TUNEL
positivity and lessened
histopathologic
deterioration
of myocardium
Ashour et al. (2012)
12 Aqueous extract 200 and
Non-insulin-dependent
diabetes mellitus
in rats
Improved insulin
sensitivity index and
blocked the rise in
homeostasis model
assessment of
insulin resistance
Anwer et al. (2008)
(continued)

Table 1. Continued.
13 Root and leaf extract
Alloxan-induced
diabetes mellitus
in rats
Normalized blood
glucose, urine glucose,
glucose-6-phosphatase,
and tissue
glycogen levels
Udayakumar et al. (2010)
14 EuMil, poly herbal
formulation 100 mg/kg
(oral route)
Chronic electroshock
stress in rats
Ameliorated cerebral
monoamine levels
Bhattacharya et al. (2002)
15 EuMil, poly herbal
formulation 100 mg/kg
(oral route)
Chronic electroshock
stress in rats
Attenuated cognitive
dysfunction,
immunosuppression,
gastric ulceration, and
plasma
corticosterone levels
Muruganandam
et al. (2002)
16 Glycowithanolides 20 and
Pentylene tetrazole
induced anxiety
in rats
Anxiolytic effects and
reduced rat brain
levels of tribulin
Bhattacharya et al. (2000)
17 Leaf extract and
Withanone 100, 200,
and 300 mg/kg
(oral route)
Scopolamine induced
toxicity in mice
Produced neuronal and
glial protection cells by
activating neuronal
proteins, oxidative
stress and
DNA damage
Konar et al. (2011)
18 Withanolide-A,
withanolides-IV,
Withanoside-VI 10
mM/kg (oral route)
Amyloid- b toxicity (rat
cortical neurons)
Promoted neurite
outgrowth, axonal,
dendritic and synaptic
rejuvenation
Kuboyama et al.
(2002, 2006)
19 Withaferin-A 1.882
mg/mouse
(intravenous route)
Human umbilical vein
endothelial cells
Inhibited TNF-a and IL-1b Ku et al. (2014)
20 Whole extract 100, 200,
and 300 mg/kg
(oral route)
6-OHDA induced
toxicity in rats
Attenuated LPO, reduced
glutathione content
and activities of
GST,GR, GPx, SOD,
and CAT
Ahmad et al. (2005)
21 Ethanolic extract
MBPQ-induced toxicity
in mice
Rescued dopaminergic
neurons, replenished
dopamine levels,
attenuated locomotor
activity, reduced
oxidative stress, and
inflammation
Prakash et al. (2014)
22 Ethanolic extract
100 mg/kg (i.p. route)
MBPQ-induced toxicity
in mice
Activated anti-apoptotic
Bcl-2 protein
expression and down-
regulated pro-
apoptotic Bax,
astrocytes,
GFAP expression
Prakash et al. (2013)
23 Standardized aqueous
extract 250 mg
(oral route)
Psychomotor
functional disorders
in healthy humans
Improved cognitive and
psychomotor
performance
Pingali et al. (2014)
24 Withanoside-IV and
Sominone 10 mM/kg
(oral route)
Alzheimer’s
disease mice
Attenuated Ab(25,35)
induced
neurodegeneration and
improved memory
deficits, prevented loss
of axons, dendrites
and synapses
Kuboyama et al. (2006)
(continued)

Somniferine, Somniferinine, and Somnine (Majumdar 1955; Maheshwari 1989;
Mishra 1989).
Pharmacological effects of Withania somnifera
Neuroprotective effect of W. somnifera
Preclinical and clinical research has supported the usage of W. somnifera for the treat-
ment of variety of neurological conditions such as cognitive disorders, anxiety, depres-
sion, senile dementia Alzheimer’s, and Parkinson’s diseases. Numerous research studies
have reported the neuroprotective effects of W. somnifera (Singh et al. 2008; Ven
Murthy et al. 2010; Wollen 2010; Durg et al. 2015; Kuboyama et al. 2014).
The antioxidant activity was demonstrated by Glycowithanolides from W. somnifera
in the cortex and striatum of rat brain by activating dose-related increase in catalase
(CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activity
(Bhattacharya and Satyan 1997). Schliebs et al. (1997) have reported the neuroprotective
activity of W. somnifera root extract. The neuroprotective activity could be due to the
presence of glycowithanolides, withanolides, and sitoindosides constituents and their
role in the inhibition of lipid peroxidation (LPO), improve CAT and GPx levels in rat
striatum and frontal cortex area. W. somnifera augment the cortical muscarinic acetyl-
choline capacity in lateral septum and frontal cortex thereby improve the cognitive
capabilities of the brain by affecting on the cortical cholinergic-signal transduction cas-
cade (Schliebs et al. 1997). Moreover, the root powder extract of W. somnifera signifi-
cantly retrieved the degenerating rat hippocampus cells exposed to immobilization
stress (Jain et al. 2001). The neurite outgrowth extensions in human neuroblastoma cell
lines were stimulated by W. somnifera root extract (Zhao et al. 2002). W. somnifera
extract also reduced kainic acid-induced excitotoxic damage (hippocampal neuronal cell
damage) by mitigating oxidative stress (Parihar and Hemnani 2003).
The streptozotocin-induced oxidative damage in treated mice was prevented by the
extract of W. somnifera by alleviating the oxidative stress (Parihar and Hemnani 2004).
In cortical neurons of rat damaged by beta amyloid protein, the extension of axons and
dendrities were done by withanolide-A, withanolide-4 and six, respectively whereas,
axonal and dendritic rejuvenation and synaptic restoration was induced by withano-
side-IV (Kuboyama et al. 2002, 2006). In the treatment of diabetic neuropathy,
Withanolide-A isolated from the roots of W. somnifera (10 mmol/kg) produces regen-
eration of neurites and reconstruction of synapses in rigorously damaged neurons
Table 1. Continued.
25 Whole extract 1 g/kg
(oral route)
Middle cerebral artery
occlusion in rats
Attenuated MDA level,
reduced lesion area
and restoration of
neurological deficits
Chaudhary et al. (2003)
26 Withaferin-A 4 mg/kg,
b.w. (intra-peritoneal)
Breast tumor
progression In
xenograft &
transgenic
mouse models
G2 and M-phase cell cycle
arrest, up-regulated
ERK/RSK axis, activation
of DR-5, Elk1 & CHOP
Nagalingam et al. (2014)

Table 2. In-vitro pharmacological activities of W. somnifera.
Sr. No. Type of extract and dose Diseased condition/model Mechanism of action References
1 Methanolic leaf extract
(2 mg/ml)
Methicillin resistant
Staphylococcus aureus
and Enterococcus spp.
Anti-bacterial activity Bisht and
Rawat (2014)
2 Methanolic extract
(0.125–2 mg/ml)
Oral infections by
Streptococcus mutans
and
Streptococcus sobrinus
Inhibited acid production,
acid tolerance and
biofilm formation of
oral bacteria
Pandit et al. (2013)
3 Withanolides (F5 and F6
fractions) (60 and 15
mg/ml)
Leishmania donovani Elevation of ROS,
Apoptosis, Cell cycle
arrest & externalization
of phosphatidylserine
Chandrasekaran
et al. (2013)
4 Glycoproteins (20 mg/ml) Aspergillus flavus,
Fusarium oxysporum, F.
verticilloides
Inhibiting hyphal growth
and spore germination
Girish et al. (2006)
5 Withaferin-A (2 mM/ml) Murine fibrosarcoma Decreased p38, ERK-1/2
and C jun JNK
Kaileh et al. (2007)
6 Withaferin-A (3 mM/ml) Cystic Fibrosis
Inflammation
(KKLEB cells)
Inhibition of IŒb
phosphorylation,
degradation &
Inetrleukin-8
Maitra et al. (2009)
7 Aqueous extract of root
powder (10 mg/ml)
Human osteoarthritis
(cartilage damage
explant models)
Inhibited gelatinase activity
of collagenase type-2
enzyme and
Decreased NO
Sumantran
et al. (2007)
8 Crude ethanolic Extract
(1 mg/ml)
Rheumatoid arthritis
(PBM cells)
Suppression of LPS-induced
production of cytokines,
interleukins and TNF-a
Singh et al. (2007)
9 Leaf extract (6, 15, 21, 25,
& 32 mg/ml)
Cancer cells (TIG1, U2OS
and HT1080)
Activation of p53,
Apoptosis pathway and
cell cycle arrest
Widodo
et al. (2008)
10 Withaferin-A (3 mM/ml) Human melanoma cells
(M14, Lu1205
and Sk28)
Promoted ROS-induced
apoptosis by lowering
Bax/Bcl2 & Bcl2/
Bim ratio
Mayola et al. (2011)
11 Withaferin-A (2 and 3
mM/ml)
Breast cancer cells (MDA-
MB-231 and MCF-7)
Translocation of Bax to
mitochondrial
membrane, Cytochrome
c release & Caspase-9
and 3 and
PARP activation
Stan et al. (2008)
12 Withaferin-A (5 and 10
mM/L)
Breast tumor progression
in xenograft and
transgenic
mouse models
G2 and M-phase cell cycle
arrest, up-regulated
ERK/RSK axis, activation
of DR-5, Elk1 & CHOP
Nagalingam
et al. (2014)
13 Withaferin-A 925 mg/ml) Human laryngeal
carcinoma Hep2 cells
Cell cycle arrest and
Blockade of
Angiogenesis
Mathur et al. (2006)
14 Withaferin-A (4 mM/ml) Human renal cancers cells PARP cleavage by down-
regulation of STAT-
3 pathway
Choi et al. (2011)
15 Withaferin-A (26 mM/ml) Renal cancers (Caki cells) Upregulation of GRP-78
& CHOP
16 Whole extract (50, 75,
and 100 mg/ml)
Coronary artery occlusion
in rats
Activated Nrf2, stimulation
of phase II detoxification
enzymes, abrogated
apoptosis in a Nrf2
Mohanty
et al. (2008)
17 Whole extract (0.15 and
0.3 mg/ml
Ab toxicity in SK-
NMC cells
Enhanced cell viability and
PPARc levels, inhibition
of acetyl-
cholinesterase activity
Kurapati et al.
(2013, 2014)
18 Water extract (0.05
and 0.1%)
Reversed glutamate-evoked
stress, restored neuronal
(continued)

Table 2. Continued.
Sr. No. Type of extract and dose Diseased condition/model Mechanism of action References
Glutamate induced
excitotoxicity in IMR-
32 and C6 cells
plasticity, reduced kainic
acid-induced excitotoxic
damage by mitigating
oxidative stress
Kataria et al. (2012)
and Parihar and
Hemnani (2003)
19 Withanolides (6.25, 12.5,
25, 50, 100 mg/ml)
Alzheimer’s disease
transgenic mice
Prevented the fibril
formation and
protection of cells from
amyloid-b toxicity
Jayaprakasam
et al. (2010)
20 Withaferin A (9.4 mg/ml) Human lung cancer cell
line (NCI-H460)
Growth inhibition and
cytotoxic activity against
human lung cancer
cell line
Choudhary
et al. (2010)
21 Withanolides (0.003 1.0
lg/ml)
Isolated rabbit jejunum Treat Alzheimer’s disease
(AD) and
associated problems
Choudhary
et al. (2005)
22 Withanolides (200 lg/mL) Isolated human
neutrophils
Treat Alzheimer’s disease
(AD) and
associated problems
Choudhary
et al. (2005)
23 Withaferin A (0, 0.156,
0.313, 0.625, 1.25,
2.5, 5 lM)
MCF-7 breast cancer cells Stimulating tumor
cell apoptosis
Zhang et al.
(2011, 2012)
24 Withaferin A (0.25, 0.5,
1.0, 1.5, 2.0 lM)
Human leukemia
U937 cells
Activation of caspase-3,
increase translocation of
cytochrome C from
mitochondria to cytosol
Oh et al. (2008)
25 Withaferin A (2 lM) Human breast cancer
cell lines
MDA-MB-231 and MCF-7
Causes G2 and M phase
cell cycle arrest
Stan et al. (2008)
26 Withaferin A (8 mg/kg) CaSki (cervical) Inhibition of Tumor growth Munagala
et al. (2011)
27 Withaferin A (0.5 lM) Human STS cell lines Anti-cancerous effect Lahat et al. (2010)
28 Withaferin A (2 mg/kg) HT-1080, SKLMS-1 (soft
tissue sarcoma)
Inhibition of Tumor growth Lahat et al. (2010)
28 Withaferin A (1.5 mM) Osteogenic sarcoma
(U2OS) and
fibrosarcoma
(HT1080) cells
Anti-cancerous effect Widodo
et al. (2008)
29 Ashwagandha leaf
extracts (0.3 mg/mL)
Human neuroblastoma
(IMR32) and rat
glioblastoma (C6) cells
Protect brain-derived cells
against oxidative stress
and induce
differentiation
Shah et al. (2015)
30 Withaferin A (8 or
12 mg/kg)
Human uveal melanoma
cell lines
Induce apoptosis Samadi et al. (2012)
31 Withaferin A (2 mM) Human umbilical vein
endothelial cells
Anti-angiogenic activity Mohan et al. (2004)
32 Withaferin A (4.0 and
5.0 mM)
Human lung cancer
cells, A549
Alters intermediate
filament organization,
cell shape
and behaviour
Grin et al. (2012)
33 Leaf extract, Withaferin A,
Withanone,
Withanolide A
(0.8 5.0 mg/ml)
Glioma cell lines C6 (rat)
and YKG1 (human)
Effective glioma therapy Shah et al. (2009)
34 Water extract of
leaves (0.12.5%)
Rat C6 glioma cell line
Human glioma
cell lines
Useful for complimentary
therapy for glioma
Kataria et al. (2011)
35 Withaferin A (5 and
10 mM)
Leukocyte-depleted
erythrocytes
Suicidal effectiveness on
erythrocyte
Jilani et al. (2013)
36 Withaferin A (2.5 to
5.0 mM)
Human breast cancer cells
(MCF-7 and
SUM159 cells)
Induct the
apoptotic process
Hahm et al. (2014)

(Kuboyama et al. 2005). Bhatnagar et al. (2009) have proved the neuroprotective
effects of W. somnifera root extract by decreasing nitric oxide (NO) production that
significantly inhibits the stress induced NADPH-diaphorase activation in the brain
through activation of cholineacetyl transferase and suppression of corticosterone
release (Bhatnagar et al. 2009). The extract obtained from leaf of W. somnifera and
its withanone constituent was found to be protective against scopolamine induced
Figure 2. Structure of main Withanolides present in W. somnifera.

toxic changes in neuronal and glial cells. The neuronal cell markers (MAP-2, NF-H,
GAP-43, PSD-95), glial cell marker, glial fibrillary acidic protein (GFAP), DNA dam-
age, and oxidative stress markers were significantly decreased by W. somnifera leaf
extract and withanone component in scopolamine induced neuronal inactivation
(Konar et al. 2011). The lead nitrate induced toxicity to brain was prevented by the
roots extract of W. somnifera that significantly increases the brain antioxidant
enzymes levels such as SOD, GPx, GST, chloramphenicol acetyltransferase and total
proteins (Sharma et al. 2011). Kataria et al. (2012) have confirmed that W. somnifera
leaf extract abolished the glutamate-induced excitotoxicity in retinoic acid-differenti-
ated C6 (glioma cell line from rat) and IMR-32 (human neuroblastoma cell line) cells
up-regulation of heat shock protein (HSP70). Furthermore, neuronal plasticity was
restored by neural cell adhesion molecules, its polysialylated form and neuronal plas-
ticity markers.
Another research study proved the neuroprotective effects of W. somnifera against
b-amyloid induced neuropathogenesis. Whereas, the in-vitro studies of W. somnifera
converses the lethal effects when human neuronal SK-N-MC cell lines were intoxicated
with b-amyloid and HIV-1Ba-L infection (Kurapati et al. 2013). The lead-induced
toxicity in glial cells was reduced by W. somnifera extract by managing the balance
of various expressions such as; neural cell adhesion molecule (NCAM), HSP70,
glial fibrillary acidic protein (GFAP), and mortalin (Kumar et al. 2014). Dar et al.
(2017) have shown that withanone-an active constituent from W. somnifera protected
NMDA-induced neuron like cells by crashing of Bax/Bcl-2 ratio, generation of
reactive oxygen species, decreasing intracellular Ca2þ
, loss of mitochondrial membrane
potential, increased expression of caspases, release of cytochrome c, attenuation
of malondialdehyde (MDA) and Poly (ADP-Ribose) polymerase-1 (Parp-1) levels which
are the indications of DNA damage (Dar et al. 2017). Dutta et al. (2018) have reported
the protective effects of W. somnifera extract in SOD1G93A mouse model of amyo-
trophic lateral sclerosis. Administration of W. somnifera extract caused significant
decrease in the levels of mutant form of SOD whereas; it enhanced the cellular chaper-
ons expression in spinal cord of SOD1G93A mice. Furthermore, it noticeably reduced
glial activation and phosphorylation of nuclear factor kappa B (Dutta et al. 2018). Birla
et al. (2019) have studied the neuroprotective effects of W. somnifera in Bisphenol A
induced-cognitive dysfunction and oxidative stress in mice. The treatment of mice with W.
somnifera improved the behavioral deficits induced by Bisphenol A whereas; the number of
Table 3. Chemical constituents present in different part of Withania somnifera.
Sr. No. Chemical constituents
1 Alkaloids Withanine, Withaninine, Pseudo-withanine, Withasomine,
Somniferine, Tropeltigloate, Somniferinine, Somninine, Nicotine,
Tropine, Pseudo-tropine, 3-a-gloyloxytropine, Choline,
Cuscohygrine, Isopelletierine, Anaferine, Anahydrine,
Scopoletin, Visamine
2 Steroids Cholesterol, b-sitosterol, Stigmasterol, Diosgenin, Stigmastadien,
Sitoinosides (VII, VIII, X, X)
3 Steroidal lactones Withaferin A, Withanone, Withanolide (E, F, A, G, H, I, J, K, L, M)
4 Flavonoids Kaempferol, Quercetin
5 Salts Cuscohygrine, Anahygrine, Tropine, Pseudotropine, Anaferine
6 Nitrogen-containing compounds Withanol, Somnisol, Somnitol

NMDA receptors in hippocampus region was restored along with anti-oxidative property by
enhancing the endogenous anti-oxidants in the brain (Birla et al. 2019) (Figure 3).
Anti-Parkinson effect of Withania somnifera
Numerous research literatures have reported a major role of W. somnifera in the
treatment of Parkinson’s disease. W. somnifera extracts have been confirmed anti-
Parkinson’s activity against functional deficits and pathological changes induced by 6-
hydroxydopamine in rat models. The study results indicated the restoration of striatal
dopamine level and its metabolites by its marked anti-oxidant activity as proved by the
diminution of LPO, GPx, GST, glutathione reductase (GR), SOD, and CAT levels.
The functional impairments like muscular coordination, locomotor activity, and drug-
induced rotational behavior were reversed due to enhancement of striatal catecholamine
level due to W. somnifera extract treatment. In addition to that W. somnifera has led to
rise in the number of surviving dopaminergic neurons as assessed by tyrosine hydroxy-
lase labeling (Ahmad et al. 2005). The treatment of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahy-
dropyridine intoxicated parkinsonian mice with root extract of W. somnifera had
maintained the anti-oxidant status by reducing the oxidant stress and therefore regu-
lated the catecholamine content in mid brain of rats. The biochemical changes such as
regulation of catecholamine level was associated with the betterment in functional activ-
ity of the rats (Sankar et al. 2007; Raja Sankar et al. 2009). Along with L-Dopa, W. som-
nifera were found to be effective in inhibiting haloperidol-induced catalepsy in mice.
Figure 3. Neuroprotective, anti-parkinsonism, and anti-alzheimer activity of W. somnifera.

Furthermore, Maneb and Paraquate induced toxicities in mouse model of Parkinson’s
disease, were diminished by ethanolic root extract of W. somnifera. There was improve-
ment in locomotor activity through anti-apoptotic, anti-inflammatory and antioxidant
effects, maintenance of tyrosine hydroxylase expression, regulation of dopamine content
of striatum nigra. W. somnifera masked the expression of inducible nitric acid synthase
(iNOS), down regulated pro-apoptotic Bax and upregulated anti-apoptotic Bcl-2 protein
expression that preceded decrease in the expression levels of GFAP (Prakash et al.
2014). Moreover, the treatment of W. somnifera root extract in rotenone induced
Parkinsonism in Drosophila melanogaster noticeably resolved the oxidative stress, mito-
chondrial dysfunctions and impaired cholinergic function by restoration of dopamine
levels and neurotransmitter functions in cerebellum and striatum of mouse brain
(Manjunath and Muralidhara 2015) (Figure 3).
Anti-Alzheimer effect of W. somnifera
Furthermore, molecular docking studies have visualized the inhibition of human acetyl
cholinesterase enzyme by withanolide-A for the treatment of Alzheimer’s diseases.
Withanoside IV and sominone-active metabolite decreased Ab(25-35) encouraged neuro-
degeneration by preventing a loss of axons, dendrites, synapses and improving memory
deficits in mice (Kuboyama et al. 2006). Computer simulation studies have proved the
ability of withanamides A and C to prevent the fibril formation and protection of cells
from Ab toxicity by binding to the active site of Ab (25-35) (Jayaprakasam et al. 2010).
The cognitive impairment and diminished acetylcholine esterase activity in rats induced
by sub-chronic exposure to propoxur-carbamate insecticide was lessened by the treat-
ment of W. somnifera (Yadav et al. 2010; Grover et al. 2012).
The root extract of W. somnifera in transgenic mice overexpressing Ab, has modified
behavioral and pathological shortfalls, plaque pathology, accumulation of b-amyloid pepti-
des and oligomers in the brains of transgenic mice besides Ab clearance by activation of
lipoprotein receptor proteins in brain blood vessels and b-amyloid peptide-degrading pro-
tease neprilysin (Sehgal et al. 2012). Furthermore, W. somnifera affords beneficial protec-
tion on cognitive deficit by improving oxidative damage produced by streptozotocin
model of cognitive impairment (Ahmed et al. 2013). The extract of W. somnifera main-
tained cell morphology in Ab-treated SK-N-MC cell lines by improving the cellular viabil-
ity by the activation of peroxisome proliferator activated receptor-c. The SK-N-MC cell
lines toxicity induced by Ab (1-42) and hydrogen peroxide recovered by the treatment of
W. somnifera by inhibition of acetyl cholinesterase activity (Kurapati et al. 2013). The
treatment of healthy human subjects with aqueous extract of W. somnifera improved the
cognitive and psychomotor activities (Pingali et al. 2014). Hence, there is great participa-
tion of W. somnifera for the treatment of Alzheimer’s disease (Figure 3).
Anti-ischemic and anti-hypoxic effect of W. somnifera
W. somnifera have demonstrated protective effect against middle cerebral artery occlu-
sion induced injury in rats by dropping oxidative stress, reduction in lesion area and
balancing the neurological functions (Chaudhary et al. 2003). The W. somnifera

treatment in mice exposed to permanent middle cerebral artery occlusion resulted in
functional recovery and diminution of infarct size. The mechanism behind decrease in
infarct size would be recovery of hemeoxygenase-1 expression and declined the upregu-
lation of proapoptotic protein PARP-1 in mouse cortex. This causes blockade of apop-
totic cascade by preventing nuclear translocation of apoptosis inducing factor. W.
somnifera also abridged the semaphorin-3A dependent inhibitory signals and thus
encouraged renovation mechanisms of dead neurons (Raghavan and Shah 2015a,
2015b). The root extract of W. somnifera and withanolide-A protected against hypobaric
hypoxia induced memory and hippocampal neurodegeneration in rats by improving
reduced glutathione levels via the activation of glutathione biosynthesis pathway in iso-
lated hippocampal cells. These effects were mediated by the activation of nuclear factor
erythroid 2-related factor two pathways and NO in a corticosterone-dependent manner
(Baitharu et al. 2013, 2014) (Figure 4).
Anti-cancer effect of W. somnifera
Over the last two decades, numerous studies have published indicating the anti-cancer
role of W. somnifera and its chemical constituents. Ashwagandha exhibits anti-cancer
activity against various types of cancer such as colon, prostate, leukemia, lung, breast,
pancreatic, renal, head and neck cancer of human cells (Yadav et al. 2010; Singh et al.
2011; Nema et al. 2013; Patel et al. 2013), stomach and skin cancer cells in mice
(Padmavathi et al. 2005). Withaferin A hampers the human umbilical vein endothelial
cells (HUVEC) proliferation by hindering the cyclin D1 expression and amending the
proteasome pathway defects (Mohan et al. 2004). The root extract of W. somnifera
killed human laryngeal carcinoma Hep2 cells via arresting the cell cycle and by
obstructing the angiogenesis process (Mathur et al. 2006). Malik et al. (2007) have
documented the anticancer activity of Withaferin-A by overexpression of tumor necrosis
factor receptor-1 and abolished the Bid expression. These anti-cancer studies declared
that withaferin-A kills the cancerous cells by apoptosis mechanism. Based on the
studies, it was decided that withaferin-A induced anticancer activity are regardless of the
participation of mitochondrial machinery (Malik et al. 2007). The W. somnifera leaf extract
Figure 4. Anti-ischemic and anti-hypoxic effect of W. somnifera.

and its phytoconstituents kills the cancer cells by different pathways such as mitogen-
activated protein kinase signaling, angiogenesis inhibition, induction of oxidative stress,
granulocyte–macrophage colony- stimulating factor signaling, p53 signaling, apoptosis
signaling, death receptor signaling and G2-M DNA damage regulation pathway (Widodo
et al. 2008).
In azoxymethane induced experimental colon cancer model in mice, it was perceived
that W. somnifera significantly altered the neutrophils, leucocytes, lymphocytes and
immunoglobulins levels. In addition to that enhanced enzymes activities such as succin-
ate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase and alpha-keto glu-
tarate dehydrogenase in colon cancerous mice leads to cell death (Muralikrishnan et al.
2010). At high concentration, aqueous extract of W. somnifera leaves induces cytotox-
icity, apoptosis and cell death of human and rat glioma cell lines (Kataria et al. 2011).
Withaferin-A demonstrated anti-cancer activity by inducing reactive oxygen species
induced apoptosis in melanoma cells by unbalancing the Bcl-2/Bax and Bcl-2/Bim pro-
teins ratio. The mechanism behind this apoptotic cascade could be activation of cas-
pases-3 and 9, release of cytochrome c into the cytosol, down-regulation of Bcl-2,
translocation of Bax to the mitochondrial membrane, abolition of trans-membrane
potential and Poly-(ADP-Ribose) Polymerase DNA fragmentation (Mayola et al. 2011).
Various mechanistic pathways were involved in the apoptosis induced by withaferin-A
and radiation in human lymphoma U937 cells such as down-regulation of Bcl-2 protein,
enhanced production of reactive oxygen species (ROS), cleavage of Poly (ADP-ribose)
Polymerase, stimulation of caspase-3, and mitogen-activated protein kinase signaling
cascade (Yang et al. 2011). Likewise, Withaferin-A also aggravated radiation induced
apoptosis in human renal cancer cells by generation of dangerous ROS, by inhibition of
Bcl-2 protein, dephosphorylation of Protein kinase B, and by endoplasmic reticulum
stress (Choi et al. 2011; Yang et al. 2011). In kidney cancer cell line, Withaferin-A
aggravated dose dependent apoptotic cell death and PARP cleavage through down-
regulation of STAT-3 pathway (Choi et al. 2011; Um et al. 2012). Vaishnavi et al.
(2012) have confirmed the use of withanone and withaferin-A for the cancer drug
development by molecular docking analysis (Vaishnavi et al. 2012). Withaferin-A sig-
nificantly introverted the development of breast cancer in transgenic mouse model by
decreasing the number of breast cancer stem cells, size, and tumor area. In the same
manner, formation of mammosphere in human breast cancer cells was blocked dose
dependently by withaferin-A treatment due to apoptosis induction and diminution of
complex-III activity (Hahm et al. 2011, 2013; Kim and Singh 2014).
Recent research studies examined that withaferin-A induces apoptosis through various
mechanisms such as, inhibiting the activation of nuclear factor kappa-B, activation of tumor
suppressor proteins, ROS generation, Par-4 induction and p38 MAP kinase activation to
induce programed cell death (Oh and Kwon 2009; Patel et al. 2013; Wadhwa et al. 2013).
Treatment of Withaferin-A inhibited the progression of breast tumor in transgenic and
xenograft mouse models that exhibited activation of Death Receptor 5, upregulation of
ERK/RSK axis and elevated levels of nuclear ETS domain containing protein-1 (Elk-1)
(Hahm and Singh 2013; Nagalingam et al. 2014). Treatment of Withaferin-A inhibits the
mammary tumors growth through the inhibition of vimentin protein expression and by
interfering with b-tubulin of cytoskeletal architecture (Antony et al. 2014; Lee et al. 2016).

The results of the various findings have discovered that, W. somnifera and its chemical con-
stituents are effective in the prevention and treatment of several kinds of cancers (Figure 5).
Anti-inflammatory effect of W. somnifera
Pronounced anti-inflammatory effect was exhibited by W. somnifera in various disease
models. Withaferin-A inhibits NFҡ -b translocation by preventing protein kinase-B
phosphorylation, degradation and gene transcription in Murine fibrosarcoma L929sA
and human embryonic kidney 293 T cells (Kaileh et al. 2007). Moreover, in human
pulmonary epithelial cells, Withaferin-A inhibited the expression of cell adhesion mol-
ecules induced by tumor necrosis factor-a (TNF- a) by inactivating the expressions of
Nuclear Factor ҡ-b (NF ҡ-b) and protein kinase-B (Oh and Kwon 2009). In cystic
fibrosis models, Withaferin-A also leads to inhibition of Interleukin-8 and NFҡ -b
(Maitra et al. 2009). In tri-nitro-benzyl-sulfonic acid induced inflammatory bowel dis-
ease model, the root extract of W. somnifera displayed anti-inflammatory and mucor-
estorative activity by improving necrosis, edema and neutrophilic infiltration in the
colon (Pawar et al. 2011). In lupus mouse model, the root powder of W. somnifera
has exhibited strong inhibitory effect on nephritis, proteinuria, various inflammatory
markers such as interleukin-6 (IL-6) and TNF-a, NO and ROS (Minhas et al. 2011,
2012). The high mobility group box-1-protein induced vascular barrier integrity in
HUVEC in mice, was protected by Withaferin-A by preventing production of IL-6,
TNF-a, expression of cell adhesion molecules, hyper-permeability, migration of
Figure 5. Anti-cancerous activity of W. somnifera.

leukocytes and activation of NFҡ-b (Lee et al. 2012). In HUVEC, phorbol-12-myris-
tate-13-acetate-induced shedding of endothelial cell protein-C-receptor was inhibited
by Withaferin-A by mitigating TNF-a and interleukin-1b (IL-1b) levels. Additionally,
Withaferin-A decreases the cecal ligation and puncture induced endothelial cell pro-
tein-C-receptor shedding in mice by decreasing the expressions of TNF-a converting
enzyme. Additionally, Withaferin-A diminished PMA-stimulated phosphorylation of
p38, extracellular regulated kinases 1/2, and c-Jun N-terminal kinase (JNK) (Ku et al.
2014). Furthermore, Withaferin-A hampers NFҡ -b activation by inhibiting TNF-a
induced expression of cell adhesion molecules by inactivation of protein kinase-B and
targeting cysteine 179 located in catalytic site of inhibitor of NFҡ-b. Withaferin-A, a
novel compound isolated from W. somnifera, enhances the cerulean induced acute
pancreatitis by reducing myeloperoxidase (MPO), nitro-tyrosine, NO levels thereby
showing the role of oxidative stress and inflammation in acute pancreatitis in animals
(Tiruveedi et al. 2018). Withaferin-A lessens the ovalbumin induced airway inflamma-
tion in mice by significantly down regulating the inflammatory cell infiltration into
the broncho-alveolar lavage fluid, pro-inflammatory cytokine expression in the lung
tissue, suppressed transforming growth factor-b1 expression, collagen I, collagen III,
a-smooth muscle actin, metalloproteinase-1, and extracellular signal related kinase 1/2
inactivation (Figure 6).
Anti-arthritic effect of W. somnifera
Abundant research work has been done on W. somnifera for the treatment of arthritis.
The ethanol extract of W. somnifera significantly inhibited lipopolysaccharide (LPS)
Figure 6. Anti-inflammatory and anti-arthritic activity of W. somnifera.

induced rheumatoid arthritis in patients by attenuating the production pro-inflamma-
tory cytokines (TNF-a and IL-1b) in peripheral blood mononuclear cells from normal
individuals and synovial fluid mononuclear cells from rheumatoid arthritis patients by
inhibiting Iҡ-b phosphorylation, nuclear translocation of NFҡ-b transcription factors
and activator protein-1. Furthermore, it also stabilized the LPS induced NO production
in RAW 264.7 cells (Singh et al. 2007). In adjuvant-induced arthritis rat model, root
powder of W. somnifera diminished the cartilage degradation by measuring the bone
collagen level (Rasool and Varalakshmi 2007). Aqueous extracts of W. somnifera root
powder have exhibited cartilage-protective effects on damaged human osteoarthritic car-
tilage through the significant inhibition of gelatinase enzyme activity (collagenase type-2
enzyme) in-vitro and prominently diminishing NO release (Sumantran et al. 2008).
Research literature suggests the important role of W. somnifera in arthritis and aids
in collagen balance by inhibition of collagenase enzyme (Ganesan et al. 2011). Whereas,
few research studies have published conflicting reports about the role of Withaferin-A
in arthritis. It has shown collagen inflammation and degradation by the up-regulation
of cyclooxygenase-2 enzyme expression through activation of microRNA-25 in rabbit
articular chondrocytes (Yu and Kim 2013; Kim and Kim 2014). Additionally, it also
exhibited increased production of intracellular ROS followed by apoptosis and increased
p53 expression via activation of JNK and PI3K/AKT pathways in rabbit articular chon-
drocytes (Yu and Kim 2013, 2014). Likewise, an administration of Ayurvedic polyherbal
formulation (BV-9238) of W. somnifera in Freund’s complete adjuvant-induced arthritis
in rats resulted in decreased production of TNF-a and NO in absence of cytotoxic
effects in rats and macrophage cell line in mouse (Dey et al. 2014). Administration of
water extract of W. somnifera root powder decreased the severity of arthritis by signifi-
cantly improving the radiological score, motor activity and decreeing the arthritic index,
auto-antibodies and C-reactive protein P levels in collagen-induced arthritis in rats
(Gupta and Singh 2014; Khan et al. 2015). Ashwashila, herbo-mineral formulation from
Indian Ayurvedic System containing aqueous extract of Ashwagandha attenuates
rheumatoid arthritis symptoms in collagen induced arthritis in mice. The Ashwashila
treatment significantly reversed the effect of C-Ab with reduced arthritis score, paw
edema, radiological and histological lesion scores joints and articular cartilage through
the inhibition of pro-inflammatory cytokines such as IL-6, TNF-a, IL-1b (Balkrishna
et al. 2019) (Figure 7).
Anti-microbial effect of W. somnifera
In the traditional system of medicine, W. somnifera has been used against variety of
microbial infections. The effectiveness of antimicrobial activity differs from microorgan-
ism to microorganism and is occurs through cytotoxicity, gene silencing, immunopoten-
tiation, etc. (Mwitari et al. 2013). In accordance with the folkloric usage, extracts of W.
somnifera have exhibited promising antifungal and antibacterial activity.
W. somnifera extracts improved the anti-bacterial effect of rifampicin and isoniazid
against E. coli and S. typhimurium (Arora et al. 2004). Increased survival rate of vital
organs of salmonellosis mice and diminished bacterial load in the same organs was
reported with W. somnifera treatment (Owais et al. 2005). The mice infected with

malaria parasite has shown significant reduction in parasite load and protection of
packed cell volume drop after the dose dependant treatment of W. somnifera with max-
imum inhibition at the dose of 600 mg/kg. Whereas, it showed non-significant inhib-
ition of chloroquine-resistant Plasmodium berghei in mice (Muregi et al. 2007). A
glycoprotein isolated from the W. Somnifera exerts fungistatic effect in different fungi
species such as Aspergillus flavus, Fusarium oxysporum, and Fusarium verticilloides by
inhibition of spore germination and hyphal growth (Girish et al. 2006). Furthermore,
W. somnifera has shown anti-leishmanial activity against free-living promastigotes and
intracellular amastigotes of Leishmania with a 50% maximum inhibitory effect (El-On
et al. 2009).
Figure 7. Anti-microbial activity of W. somnifera.

Many gram-negative bacteria such as Salmonella typhi, Escherichia coli, Citrobacter
freundii, Proteus mirabilis, Klebsiella pneumonia, and Pseudomonas aeruginosa were
inhibited by the extracts of W. somnifera confirming its antibacterial activity (Singh
and Kumar 2011; Alam et al. 2012). Furthermore, the flavonoids isolated from W.
Somnifera have demonstrated pronounced anti-fungal effect against Candida albicans
with 0.039 minimum inhibitory concentration and 0.039 mg/ml minimum fungicidal
concentration. However, it was found to be ineffective against Aspergillus niger and
Aspergillus flavus fungal species (Singh and Kumar 2011). W. somnifera also exhibits
potent in-vitro anti-bacterial activity against Salmonella typhimurium (Alam et al.
2012). Withanolides causes in-vitro apoptotic death in Leishmania donovani by pro-
voking cell cycle arrest at G0/G1 phase, DNA breakage, externalization of phospha-
tidyl serine, decrease in mitochondrial potential via increasing formation of ROS and
blocking the protein kinase-C signaling pathway (Grover et al. 2012; Chandrasekaran
et al. 2013). The acid production, acid tolerance and formation of biofilm of oral
bacteria, Streptococcus sobrinus and Streptococcus mutans was prevented by W. som-
nifera at minimum inhibitory concentration levels (Pandit et al. 2013). The protective
action of W. somnifera was potentiated in L. donovani-infected mice treated with cis-
platin as compared to donovani-infected mice treated with W. somnifera only by
expressing the concentration of T cells and markers of natural killer cell (Sachdeva
et al. 2013). The methanolic extract of W. somnifera leaves has revealed noticeable
anti-bacterial activity against gram-positive isolates obtained from pus samples of
methicillin-resistant Staphylococcus aureus and Enterococcus spp. (Bisht and Rawat
2014) (Figure 7).
Anti-stress and adaptogenic effect of W. somnifera
W. somnifera is a recognized stress relieving herbal remedy being used in Ayurvedic
medicine as an anti-stress medication. It resulted in enhanced stress tolerance in ani-
mals as well as human beings (Bhattacharya et al. 1995; Kaur et al. 2001; Singh et al.
2001). The result reported by Archana and Namasivayam 1999 has indicated the anti-
stress property of W. somnifera in cold water swim stress (Archana and Namasivayam
1999). The similar results were reported by Rege et al. (1999) thus showing the anti-
stress and adaptogenic activity of W. somnifera (Rege et al. 1999). Glycowithanolides
obtained from W. somnifera exhibited an anxiolytic effect against pentylene tetrazole
induced anxiety in rats. In addition to that, it decreased the level of endocoid marker (trib-
ulin) of clinical anxiety and mitigated oxidative stress induced LPO in the frontal cortex
and striatum of chronic foot shock stressed rats (Bhattacharya et al. 2000, 2001). Dhuley
(2000) has stated the adaptogenic potential of W. somnifera in frogs and rats (Dhuley
2000). Another research study has indicated the anti-stress effect of W. somnifera root
extract in stress-induced neuronal degeneration in rats. In hippocampal sub-layer, the ultra-
structural study of neuronal cell bodies was performed that indicates the cytoprotective
effect of W. somnifera in improving the neuronal degenerating features such as membrane
blebbing, chromatin condensation and fragmentation, karyorrhexis and intracellular spacing
in rat brain (Shukla et al. 2000). Additionally, a poly herbal formulation of W. somnifera
(EuMil) markedly improved the cerebral monoamine neurotransmitter levels such as nor-

adrenaline, dopamine and 5-hydroxytryptamine that were induced by chronic electroshock
stress (Bhattacharya et al. 2002). Moving ahead, EuMil also normalized chronic stress-
induced glucose intolerance, male sexual behavior and frustration along with retrieval of
immunosuppression, gastric ulceration, cognitive dysfunction and plasma corticosterone lev-
els (Muruganandam et al. 2002). Chronic Fatigue Syndrome (CFS) is a disorder considered
by persistent and relapsing fatigue. The anti-stress activity of W. somnifera was tested in
CSF in mouse model. In this study the fatigue was induced by forcing the mice to swim
for six minute per day for continuous 15days. The treatment of mice with W. somnifera
before the stress exhibited a significant reduction in immobility time showing the anti-stress
activity of W. somnifera due to its antioxidant property. The induction of chronic fatigue
by forced swimming for 15days caused a significant increase in the brain MDA levels as
compared to naïve mice representing the oxidation of proteins, lipids and DNA. Treatment
of mice with W. somnifera extract at a dose of 100mg/kg, p.o. resulted in significant rever-
sal of LPO (Singh et al. 2002). Thus, W. somnifera is added in diverse therapeutic prepara-
tions for the treatment of stress and related disorders (Muruganandam et al. 2002). The
adaptogenic activity of root extract of W. somnifera was tested against chronic stress
induced in Wistar rat model. The chronic stress induced significant increase in blood glu-
cose level, increase in plasma corticosterone levels, gastric ulcerations, cognitive deficits,
male sexual dysfunction; immunosuppression and mental depression were reversed by the
treatment of W. somnifera root extract thereby confirming its anti-stress adaptogenic activ-
ity (Bhattacharya and Muruganandam 2003). The administration of aqueous extract fraction
of W. somnifera roots in mice, showed reduction of T-cell population and upregulated Th1
cytokines (Khan et al. 2006). Perment is another poly-herbal formulation of W. somnifera
revealed significant anti-depressant and anxiolytic activity in rats that was due to partial
stimulation of adrenergic and serotonergic systems (Ramanathan et al. 2011). In a clinical
study, the root extracts of W. somnifera controlled the serum cortisol levels devoid of any
side effects in human subjects (Chandrasekhar et al. 2012) (Figure 8).
Figure 8. Anti-stress and adaptogenic activity of W. somnifera.

Anti-diabetic effect of W. somnifera
W. somnifera is one of the important herbs of Indian systems of medicine having
anti-diabetic activity. Numerous Ayurvedic polyherbal formulations such as Dianix,
Trasina exhibited blood sugar reducing activity in human subjects (Bhattacharya et al.
1997; Mutalik et al. 2005; Gauttam and Kalia 2013). Moreover; oral administration of
W. somnifera root powder for 30days controlled the blood sugar level that was comparable
to an oral anti-diabetic drug daonil (Andallu and Radhika 2000). The treatment of rats
with aqueous W. somnifera extract significantly stabilizes the hyperglycemia in non-
insulin dependent diabetes mellitus by improving insulin sensitivity index (Anwer
et al. 2008).
The treatment of rats with W. somnifera root and leaf extracts showed significant
normalization of blood glucose, urine glucose, glucose-6-phosphatase and tissue
glycogen levels in alloxan-induced diabetes mellitus via enzymatic and non-enzymatic
antioxidant mechanisms (Udayakumar et al. 2009, 2010). Withaferin-A also obstructs
the inflammatory response in cytokine induced damage to pancreatic islets in in-vitro
and successive transplantation (SoRelle et al. 2013). W. somnifera leaf and root
extracts exhibited anti-diabetic activity in a dose-dependent manner by improving
the glucose uptake in adipocytes and skeletal myotubes. The leaf extract of W. somni-
fera showed more prominent effects than the root extract of W. somnifera (Gorelick
et al. 2015). Withaferin-A mitigates multiple low doses of Streptozotocin induced
type 1 diabetes mellitus in rats. Withaferin-A improved the Streptozotocin induced
oxidative and nitrosative stress along with significant reduction in the pro-inflamma-
tory cytokines levels such as TNF-a and IL-6, DNA fragmentation and apoptosis,
supporting the protective effect of Withaferin-A in diabetes mellitus (Tekula et al.
2018) (Figure 9).
Aphrodisiac effect of W. somnifera
Low sperm count and reduction of testosterone hormone levels are signs of defective
spermatogenesis process that has indicated the physiological problems of Sertoli and
Leydig cells (Sharpe 1993). The treatment of 25 days old female rats with W. somnifera
extract exhibited significant changes in gonadotropin hormone levels along with
improvement of ovary weight and folliculogenesis process (Al-Qarawi et al. 2000).
The treatment of Wistar rats with W. somnifera extracts was found to decreased testes
weight and improved number and diameter of testicular seminiferous tubular cell.
Therefore, W. somnifera exhibited testicular development and spermatogenesis process
by lowering the testosterone and follicle-stimulating hormone levels and increasing
the interstitial cells stimulating hormone levels (Abdel-Magied et al. 2001).
Additionally, a research report was published indicating the warned use of W. somni-
fera against sexual incompetence. In this study, administration of W. somnifera
resulted in significant decrease in sexual performance, sexual vigor, libido, penile erec-
tion, and these effects were reasonably reversed by the cessation of W. somnifera treat-
ment. These anti-masculine effects were not due to alterations in testosterone levels or
toxicity but might be associated with serotonergic, hyperprolactinemic, GABAergic or
sedative activities of the extract (Ilayperuma et al. 2002). In a different study of

infertility in men, the extract of W. somnifera regularized the sperm count and motil-
ity by decreasing the follicle-stimulating, testosterone, luteinizing and prolactin hor-
mones serum levels (Ahmad et al. 2010). Furthermore, W. somnifera also reported
non-significant enhancement of psychogenic erectile dysfunction management
(Mamidi and Thakar 2011). The treatment with W. somnifera in sexually lethargic
mice has resulted in enhanced sperm production and improved blood testosterone lev-
els. The extract of W. somnifera was found to decrease the cadmium toxicity on motil-
ity and density of cauda epididymidal sperm and seminiferous tubules (Mishra et al.
2012). The treatment of oligospermic patients with 675 mg of W. somnifera extract
per day for 90 days resulted in increased sperm count, semen volume and sperm
motility by improving testosterone and serum luteinizing hormone levels (Ambiye
et al. 2013) (Figure 10).
Cardio-protective effect of W. somnifera
W. somnifera possesses cardio-tropic and cardio-protective activity in preclinical and
clinical models (Das et al. 1964; Prince et al. 2008; Ojha and Arya 2009). Polyherbal
formulations containing W. somnifera as one of the constituent revealed cardio-pro-
tective activity in animals by activation of nuclear factor-erythroid-2-related
Figure 9. Anti-diabetic activity of W. somnifera.

transcription factor (Nrf)2, phase-II detoxification enzymes, and abrogating apoptosis
process (Mohan et al. 2006; Thirunavukkarasu et al. 2006; Reuland et al. 2013). The
prophylactic treatment with W. somnifera extract significantly maintained the balance
between oxidant and antioxidant (Gupta et al. 2004; Mohanty et al. 2004; Ashour
et al. 2012), decreased histopathological damage of rat myocardium, anti-apoptotic
activity measured by using terminal deoxynucleotidyl transferase dUTP nick-end label-
ing (TUNEL) assay in coronary artery occlusion rat model (Mohanty et al. 2008).
Likewise, the standardized extract of W. somnifera inhibited doxorubicin induced car-
dio-toxicity by decreasing the biochemical changes of rat myocardium (Hamza et al.
2008). The oral pretreatment of rats with W. somnifera extract (100 mg/kg) 4 weeks
elicited a significant cardio-protective activity by lowering cardiac troponin-I level,
LPO level, lipid profiles, and cardiac marker enzymes along with elevation of antioxi-
dant enzymes in isoproterenol induced oxidative damage in rat myocardium. The
probable mechanism of cardio-protection activity would be augmentation of the
endogenous antioxidant system and an inhibition of LPO in the rat myocardial mem-
brane (Khalil et al. 2015) (Figure 11).
Figure 10. Aphrodisiac activity of W. somnifera.

Hepato-protective effect of W. somnifera
Several research studies were implemented to evaluate the hepatoprotective potential
of W. somnifera. In one of study, W. somnifera has exhibited hepatoprotective activ-
ity against gamma radiation induced toxicity in rodents. The treatment of rodents
with 100 mg/kg dose of W. somnifera significantly reduces the MDA levels, hemeoxy-
genase levels, hepatic serum enzymes, and total nitrate/nitrite activity in liver.
Whereas, the levels of serum antioxidant enzymes such as GPx and SOD were found
to be elevated in hepatic tissues (Hosny and Farouk 2012). Furthermore, the treat-
ment of rats with W. somnifera at a dose 500 mg/kg significantly decreases the
increased biomarkers levels such as alanine transaminase (ALT), aspartate amino-
transferase (AST), alkaline phosphatase (ALP), bilirubin when treated with hepato-
toxic dose of paracetamol. It also significantly reduces the LPO, enhances CAT,
glutathione content, GPx and GR activity in liver (Malik et al. 2013; Sabina et al.
2013). The hepatoprotective activity of withanolide-rich fraction isolated from a
methanolic extract of Withania somnifera roots were performed in acetaminophen
(750 mg/kg, p.o. for 14 days) intoxicated rats. The withanolide-rich fraction group
have shown significant decrease in serum bilirubin, ALP, AST and ALT levels
whereas, significant increase in hepatic SOD, GPx and total antioxidant capacity. The
MDA and NO levels were found to be significantly decreased along with down
Figure 11. Cardio-protective activity of W. somnifera.

regulation of mRNA expression of TNF-a, IL-1b, cyclooxygenase-II, and iNOS genes
(Devkar et al. 2016). Above research outcomes, support the usage of W. somnifera
for the treatment of many hepatic ailments (Figure 12).
Immunomodulatory effect of W. somnifera
Immunomodulatory activity of W. somnifera has exhibited a significant modulation of
immune response in animal models. These observational studies suggest that W. somni-
fera can be used as an immunological adjuvant with several therapeutic benefits in
AIDS, cancer, and infection (Grandhi et al. 1994). The myelosuppression in mice
induced by three immunosuppressive drugs (azathioprine, cyclophosphamide and pred-
nisolone) was prevented by treatment of mice with W. somnifera extract. Treatments
with W. somnifera extract exhibited a significant increase in the red blood cell count,
platelet count, hemoglobin count and body weight in mice (Ziauddin et al. 1996). W.
somnifera extract significantly reduced the leukopenia condition induced by cyclophos-
phamide treatment. The numbers of b-esterase positive cells in the bone marrow of
cyclophosphamide treated animals were also found to be increased after W. somnifera
extract treatment (Davis and Kuttan 1998). W. somnifera extract significantly reduced
the leukopenia condition induced by sub-lethal dose of gamma radiation (Kuttan 1996).
The extract of W. somnifera were studied for cyclophosphamide induced immunosup-
pression in mice by using two models such as active paw anaphylaxis and delayed type
hypersensitivity. The results of the study showed the cyclophosphamide induced
potentiation of delayed type hypersensitivity reaction was suppressed in animals treated
Figure 12. Hepatoprotective protective activity of W. somnifera.

with W. somnifera extract along with significant increase in white blood cell (WBC)
counts and platelet counts (Agarwal et al. 1999). Treatment of Balb/c mice with W.
somnifera root extract (20 mg/kg, i.p.) was found to stimulate the immunological activ-
ity. Treatment of mice with five doses of W. somnifera root extract enhanced the WBC,
bone marrow cellularity and alpha-esterase positive cell number, circulating antibody
titer and number of plaque forming cells in the spleen. The extract also inhibited
delayed type hypersentivity reaction (Mantoux test) and improved phagocytic activity of
peritoneal macrophages in mice (Davis and Kuttan 2000). In a recent study, a polyher-
bal formulation (Immun-21), containing W. somnifera as one of the constituent, exhib-
ited significant immunomodulatory activity expressing its clinical benefits in human
immuno-deficiency virus (HIV) patients (Singh et al. 2001). Prakash et al. (2002) have
reported the cytoprotective activity of W. somnifera against experimental skin cancer in
which the levels of SOD, glutathione, GPx and CAT were normalized following the
administration of W. somnifera (Prakash et al. 2002). Numerous research studies have
demonstrated the immuno-stimulatory potential of W. somnifera, resulting in enhanced
phagocytic activity of macrophages, hemolytic titers and inhibition of delayed type sen-
sitivity. In normal and tumor containing mice, W. somnifera treatment had shown a
positive effect on natural killer cells resulting in improved cell killing (Davis and
Kuttan 2002).
W. somnifera has exhibited immuno-modulatory effects on cytotoxic lymphocyte pro-
duction leading to reduced tumor growth. Withaferin-A has exhibited better immuno-
modulatory activity than doxorubicin in inhibiting growth of breast and colon cancer
cell lines (Jayaprakasam et al. 2003). The cytoprotective potential of W. somnifera was
examined by Diwanay et al. (2004). In that study W. somnifera has exhibited myelopro-
tection in tumor model without conceding anti-tumor efficacy of azathioprine, cyclo-
phosphamide or prednisolone (Diwanay et al. 2004). Withaferin A and Withanolide E
exhibited specific immunosuppressive effect on human T and B lymphocytes and on
mice thymocytes. Withanolide E had specific effect on T lymphocytes whereas
Withaferin A affected both T and B lymphocytes (Aggarwal et al. 2012). In the present
study, W. somnifera were tested as an immunomodulatory drug against doxorubicin
induced immunosuppression study. The treatment of rats with W. somnifera extract
exhibited significant increase in the white blood cells, absolute lymphocyte and platelets
count after bone marrow suppression induced by doxorubicin. Bone marrow cellularity
as well as alpha-esterase positive cell number also increased significantly. Treatment
with W. somnifera extract along with antigen produced an enhancement in the circulat-
ing antibody titer and number of plaque forming cells in the spleen (Rizvi et al. 2016).
The immunomodulatory of W. somnifera extract and Withaferin A supplementation
was reported on zinc oxide nanoparticles mediated toxicity in Balb/c mice. The animals
were exposed to zinc oxide nanoparticles along with W. somnifera extract and
Withaferin A for 28 days and different parameters such as, body weight, organ coeffi-
cient, cytotoxicity, total serum protein levels, NO, phagocytosis, TLR6 and arginase gene
expression were measured. The toxicity of zinc oxide nanoparticles was found to be
reduced in presence of W. somnifera extract and Withaferin A with decreased TLR6
and arginase gene expression and restoration of phagocytic activities in Balb/c mice
(Kumar et al. 2019) (Figure 13).

Clinical evaluation of W. somnifera
Effect of W. somnifera on insomnia and anxiety
Langade et al. (2019) have examined the safety and efficacy of W. somnifera root extract
in insomnia and anxiety. The result of the study shown that, sleep onset latency, sleep
efficiency, Pittsburgh Sleep Quality Index, and anxiety scores with W. somnifera root
extract treatment for ten weeks in test groups when compared to placebo group. So, it
can be concluded that W. somnifera plant is of potential use to enhance sleep in insom-
nia and anxiety patients (Langade et al. 2019).
Effect of W. somnifera on memory and cognition
Choudhary et al. (2017) have studied the safety and efficacy of W. somnifera root extract in
improving memory and cognitive functions. The W. somnifera treatment group demon-
strated significant improvements in both immediate and general memory as evidenced by
Wechsler Memory Scale III subtest scores as compared to placebo group at the end of eight
week study. The treatment group also demonstrated significant improvement in executive
function, sustained attention and information-processing speed as indicated by scores on
the Eriksen Flanker task, Wisconsin Card Sort test, Trail-Making test part A, and the
Mackworth Clock test. Hence, W. somnifera is effective in enhancing both immediate and
Figure 13. Immunomodulatory effect of W. somnifera.

general memory in people with mild cognitive impairment as well as improving executive
function, attention and information processing speed (Choudhary et al. 2017).
Effect of W. somnifera on stress and weight management
Choudhary et al. (2017) have examined the effect of W. somnifera root extract on body
weight management under chronic stress in adults. At the end of experiment of W.
somnifera exhibited significant reduction in PSS scores, reduced cravings for food as
compared to placebo group. The Food Cravings Questionnaire (FCQ) scores were
reduced significantly in the W. somnifera treated group. Significant decrease in serum
cortisol levels, body weight and Body Mass Index was found with W. somnifera treat-
ment group. Hence, it can be concluded that W. somnifera root extract can be used for
body weight management in adults under chronic stress (Choudhary et al. 2017).
Effect of W. somnifera on thyroid gland function
The safety and efficacy of W. somnifera root extract was studied by Sharma et al. (2018)
on thyroid gland function. The eight weeks treatment of W. somnifera root extract in
hypothyroid patients significantly improved the serum thyroid stimulating hormone, tri-
iodothyronine and thyroxine levels as compared to placebo (Sharma et al. 2018).
Effect of W. somnifera on telomerase activity
The telomerase activity of W. somnifera root extract was studied on human HeLa cell
line by Raguraman and Subramaniam in 2016. The result of the study showed that W.
somnifera root extract enhanced telomerase activity through telomeric repeat amplifica-
tion protocol assay with highest enhancement of 45% at 10–50 lg concentration. So, it
is concluded that W. somnifera root extract exhibits the anti-aging potential
(Raguraman and Subramaniam 2016).
Effect of W. somnifera on cardio-respiratory endurance
Choudhary et al. (2015) have studied the effect of W. somnifera in improving cardio-
respiratory endurance in healthy athletic adults. There was a significant increase in the
oxygen consumption at peak physical exertion level (VO2 max) at 8 (4.91) and 12
(5.67) weeks after the treatment with W. somnifera root extract as compared to placebo
group with improvement in the quality of life scores. It is concluded that, the root
extract of W. somnifera enhances the cardiorespiratory endurance and improves quality
of life in healthy athletic adults (Choudhary et al. 2015).
Effect of W. somnifera on testosterone, muscle strength, and recovery
Wankhede et al. (2015) have studied the effect of W. somnifera on muscle strength and
recovery. The group treated with W. somnifera exhibited significant increase in muscle
strength (bench-press and leg-extension exercise), muscle size increase at the arms and

chest, testosterone level, reduction of exercise induced muscle damage and body fat per-
centage when compared with placebo group (Wankhede et al. 2015).
Effect of W. somnifera on female sexual function
The safety and efficacy of W. somnifera root extract was tested in improving sexual
function in women by Dongre et al. (2015). The treatment of females with high concen-
tration ashwagandha root extract leads to significant improvement in the Female Sexual
Distress Scale score and Female Sexual Function Index score for arousal, lubrication,
orgasm and satisfaction and the number of successful sexual encounters at the end of
treatment. This indicates that, the treatment of W. somnifera root extract improves the
sexual function in healthy women (Dongre et al. 2015).
Effect of W. somnifera on testosterone male sexual function
Ambiye et al. (2013) have studied the clinical evaluation of the spermatogenic activity
of W. somnifera root extract in Oligospermic Males. The results of the study showed
that there was 167% increase in sperm count, 53% increase in semen volume, 57%
increase in sperm motility and serum testosterone levels on day 90 as compared to pla-
cebo-treated group. So, from the Ayurveda point of view, W. somnifera indicated in the
treatment of oligospermia leading to infertility (Ambiye et al. 2013).
Effect of W. somnifera on anti-Aging activity
Kumar et al. (2013) have studied the anti-aging property of W. somnifera root extract
Treatment of Caenorhabditis elegans round worm with W. somnifera root extract has
extended the round worm lifespan by 20% (Kumar et al. 2013).
Effect of W. somnifera on stress and anxiety
Chandrasekhar et al. (2012) have conducted the randomized double-blind, placebo-con-
trolled study of W. somnifera root extract in reducing stress and anxiety in adults. The
results of the study exhibited that treatment group displayed significant reduction in
scores on all stress-assessment scales on day 60 as compared to placebo group along
with significant decrease in serum cortisol levels and absence of serious side effects. So,
from the study it was clear that W. somnifera root extract safely and effectively
improves an individual’s resistance toward stress and thereby improves self-assessed
quality of life (Chandrasekhar et al. 2012).
Safety and toxicity profile of W. somnifera
The leading step before therapeutic usage of any new medicinal herb is the assessment
of its possible toxicity on various body systems (Mishra et al. 2000). Numerous research
studies have reported the safety and efficacy of W. somnifera extract were found to be
safe for all the age groups, males and females and even during the pregnant condition

Table 4. Marketed formulation of W. Somnifera along with their applications.
Sr. No. Product name Company name Applications of product
1 Stresscom Dabur India Ltd. Relieves anxiety, Neurosis, Mental
stress, Depression
2 Ashvagandha Morpheme Remedies Combating stress
3 Stresswin Baidynath Ayurved Bhawan Combating exertion, Reduction in
anxiety, Strain, Stress,
Mental alertness
4 Himalaya Massage oil The Himalaya Drug Co. Relief from stress and insomnia
5 Brento Zandu Pharmaceutical
Works Ltd.
Nerve tonic
6 Ashwagandha Ayurceutics Stress reliever
7 Ashwagandharista Baidynath Ayurved Bhawan Nerve tonic, Memory, Cognition
improvement, Natural sleep inducer
8 Dabur Ashwagandha Churna Dabur Combating stress
9 Arshadi pills Dehlvi Remedies Stress, Depression, Cardiac tonic
10 100% Natural W.
somnifera extract
Xi’an Saina Biological
Technology Co., Ltd
Anti-allergy, Anti-histamine, Anti-
pyretic, Pain-relieving,
Local anesthetic
11 Ashwagandha/W.
somnifera extract
Wuxi Gorunjie Natural-Pharma
Co., Ltd.
Anti-inflammatory, Anti-arthritic,
Stress reliever
12 Ashwagandha extract Nanjing Zelang Medical
Technology Co., Ltd.
Anti-allergy, Anti-histamine, Anti-
bacterial, Local anesthetic, Anti-
pyretic, Anti-inflammatory,
Rheumatoid arthritis, Improves
sexual function
13 Nutramax W. somnifera
extract (10:1).
Hunan Nutramax Inc Adaptogen, Anti-stress agent,
aphrodisiac, Growth promoter,
Hypotensive, Anti-spasmodic,
Respiratory stimulant, Sleep inducer
14 Natural 80 mesh American
ginseng root extract
Qingdao Fraken International
Trading Co., Ltd.
Anti-tumor, Anti-arthritic, Anti-
inflammatory, Immunosuppressive
15 Nutramax-AE (100%). Hunan Nutramax Inc Anti-allergy, Anti-histamine, Anti-
pyretic, Pain-relieving,
Local anesthetic
16 Amaybion Aimil Pharmaceuticals Pvt. Ltd Anti-ageing
17 Amry-gel Aimil Pharmaceuticals Pvt. Ltd Health-Tonic
18 Ashwagandh tablets Himalyan Drugs Co. Anti-stress
19 Articulin-F Eisen Pharmaceuticals Pvt. Ltd Useful in rheumatoid Arthritis
20 Aswal Plus Gufic Ltd. Ageing, Anti-stress, Adaptogenic,
Rejuvenating agent
21 Geriforte Himalyan Drugs Co. Health supplement, Physical and
mental exhaustion, Anxiety, Post-
menopausal syndrome
22 Gestone Zandu Pharmaceuticals
works Ltd
Preeclampsia, Placental insufficiency,
Threatened abortion, and
pregnant anemia
23 One Be Lupin Lab Ltd. Stress, Ageing, Adaptogen,
Rejuvenator, Immuno modulator
24 Imunocin Nukem Remedies Ltd. Immunomodulator
25 Stir Targof Pure Drug Ltd. Stress, Male subfertility, Oligospermia,
Adaptogen, Neurostimulator
26 Stresscom Dabur India Ltd. Anti-stress, Anxiety,
Depression, Insomnia
27 Keshari Kalpa Baidyanath Ayurved Bhavan Ltd. Aphrodisiac, Health tonic
28 Lactare TTK Pharma Pvt. Ltd. Improved lactation
29 Lacton Baidyanath Ayurved Bhavan Ltd. Improved lactation
30 Medispermina Mesi Products (p) Pvt. Ltd. Azoospermia, Oligospermia,
Impotence, Infertility
31 Mustong TTK Pharma Pvt. Ltd. Revitaliser
32 Sioton Albert David Ltd. Improved libido, Sexual fatigue
33 Vigorex SF Zandu Pharmaceuticals
works Ltd
Stress reliever, Improves strength

(Sharma et al. 1985). The acute and sub-acute oral toxicities was performed in Wistar
rats by using hydro-alcoholic root extract of W. somnifera at a dose of 2,000 mg/kg/
body weight and was found to be safe. The extract was administered at 2,000 mg/kg for
acute toxicity study and animals were monitored for 14 days whereas, for sub-acute tox-
icity the extract was administered at different doses such as 500, 1,000, and 2,000 mg/kg
and observed for 28 days. No noticeable changes were observed in the body and organ
weight and hematological parameters in the rats (Prabu et al. 2013). Moreover, the
extract of W. somnifera was found to be safe for developing rat fetuses and pregnant
mothers including death, structural deformities and body growth but no apparent
changes were found in the pregnant mother or in the fetus (Prabu and Panchapakesan
2015). Once daily intraperitoneal injection of W. somnifera extract at 1,100 mg/kg dose
in Swiss albino mice did not revealed any death within 24 h of injection. But, a little
increase in dose causes death with an LD50 of 1,260 mg/kg/body weight in acute toxicity
study. No changes were observed in peripheral blood constituents. However, significant
reductions were found in the spleen, thymus, and adrenal gland weights (Sharada et al.
1993; Tiwari et al. 2014). Hence, W. somnifera can be used as safe medicinal herb for
the treatment of various diseased conditions.
Marketed formulations of W. somnifera
Ashwagandha is advocated as a protective drug against atherosclerosis, hypertension,
and coronary heart diseases. It reduces the sensitivity of the heart to adrenergic stimula-
tion and thereby protects the heart against sympathetic outbursts. Moharana (2008)
reported that the roots and leaves of Ashwagandha are traditionally used in the form of
powder, decoction, or oil. These have been used in folk medicine against general dis-
ability, hypertension, inflammation, and wounds. Thirunavukkarasu et al. (2006) found
Ashwagandha to have energy boosting properties and recommended its use as a dietary
supplement for cardioprotection. The effect of Ashwagandha root was evaluated for
lipid peroxidation in stress. The herb was found to have very good antioxidant activity,
which may partly explain the anti-stress, congestion-facilitating, anti-inflammatory, and
anti-aging effects of this herb (Moharana 2008; Table 4).
Conclusion
W. somnifera is a medicinal plant with recognized ethnopharmacological and pharma-
ceutical properties. It has numerous clinical applications in Indian Systems of Medicine.
In pre-clinical studies, W. somnifera and its chemical constituents had exhibited several
pharmacological properties such as anti-oxidant, aphrodisiac, anti-microbial, adapto-
genic, diuretic, anti-helminthic, immunomodulatory, anti-inflammation, anti-diabetic,
hepatoprotection, anti-stress, anti-ulcer, anti-rheumatic, oligospermia, neuro-protection,
anti-apoptotic, angiogenic, inhibition of NFŒ-b transcription, MAPK signaling pathways,
and endoplasmic reticulum stress reducing effects. Clinically, W. somnifera has proven
for its Insomnia, anxiety, memory stress, weight management, thyroid gland function,
telomerase, cardio-respiratory endurance, muscle strength, recovery of male and female
sexual function, and anti-aging activity. Toxicity studies of W. somnifera has not

indicated any toxicity or side effects till date and thus can be safely used in human
beings for the treatment of acute and chronic diseased conditions. Due to the presence
of variety of bioactive constituents, their low toxicity and unique mechanism of action
makes W. somnifera a suitable drug candidate for the treatment of various diseases.
However, there is an urgent need of research at the phytochemistry and pharmacology
interface toward W. somnifera drug development with increased pharmacological activ-
ity and minimal toxicity as compared to its available commercial formulations.
Acknowledgments
The authors want to acknowledge Poona college of Pharmacy, Pune for online article support.
Conflicts of interests
The author has no conflicts of interest to declare.
About the authors
Dr. Deepa S. Mandlik (Ingawale) holds PhD degree in Pharmacology and working as an
Assistant Professor in Poona College of Pharmacy. Bharati Vidyapeeth Deemed to be University,
Pune. Her research areas are Inflammation, Immunity, Anticancer, Gene expression studies,
Anti-diabetic and Hepatoprotection.
Dr. Ajay G. Namdeo holds PhD degree in Pharmacognosy and working as an Associate
Professor in Poona College of Pharmacy. Bharati Vidyapeeth Deemed to be University, Pune. His
research areas include natural products development.
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