Evaluation of antidiabetic and antiinflammatory activity of fatty extract of Caprus hircus milk in rats

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__________________________________________
* Corresponding author: Shakti Prasanna Sahoo
E-mail address: sahoo.shakti@gmail.com
Available online at www.ijrpp.com
Print ISSN: 2278 – 2648
Online ISSN: 2278 - 2656 IJRPP | Volume 2 | Issue 2 | 2013 Research article
Evaluation of antidiabetic and antiinflammatory activity of fatty extract of
Caprus hircus milk in rats
1
D.N.Anusha , 1,
*Shakti Prasanna Sahoo, 1
S.Ramchandran, 1
M.D.Dhanraju and
2
P.K.Kota
Department of Pharmacology, GIET School of Pharmacy, Chaitanya Knowledge City,
Rajahmundry, 533296, Andhra Pradesh, India.
2
Delve Labs Pvt. Limited Balanagar, Hyderabad.
ABSTRACT
Capra hircus(goat) is one of the domesticated animals used for milk, meat and fibre. Milk is one of the essential
products in the human diet, rich in nutritive components. Goat's milk has vitamins, minerals, trace elements,
electrolytes, enzymes, proteins, and fatty acids that are easily assimilated by the body. The fatty acids present in
Caprus hircus milk was revealed to have more beneficial properties to health and used as a traditional medicine
for.The present study is aimed to investigate the antidiabetic and anti-inflammatory effect of isopropanol extract
of milk of Caprus hircus. The antidiabetic activity was evaluated by measuring blood glucose level in rats
treated with isopropanol extract of caprine milk and comparing to normal and streptozotocin (STZ) induced rats.
The isopropanol extract of Caprus hircus produced significant changes in the streptazotocin induced diabetic
rats by reducing the glucose levels considerably. isopropanol extract treated group(200 mg/kg b.w) showed the
mean value of 157.6±3.656 and 117.6±3.839 respectively as compared with a mean value 317.2±4.244 of
diabetic control group. The anti-inflammatory activity was assessed by carrageenam induced paw edema model
in rats.There was significant reduction (p< 0.01) in paw diameter in the group that received high dose (200
mg/kg b.w) of isopropanol extract of Caprus hircus milk from Mean±SEM of 0.184±0.004 to 0.052±0.003
(71.73%) as compared with the disease control group. The present study concludes that caprine milk extract
confirmed promising anti diabetic activity in streptozotocin induced diabetic Wister rats and significant anti-
inflammatory properties against carrageenan induced paw edema.
Keywords: Caprus hircus, fatty acid, antidiabetic, stretozotocin, inflammation, carrageenam, paw edema.
Introduction
Diabetes is a metabolic disorder of multiple
etiologies characterized by chronic hyperglycemia
with disturbance of carbohydrate, fat, and protein
metabolism resulting from defects in insulin
secretion, insulin action, or both. The disease is
associated with microvascular, macrovascular, and
metabolic complications.When the renal threshold
for glucose reabsorption is exceeded, glucose spills
over into the urine (glycosuria) and causes an
osmotic diuresis (polyuria), which, in turn, results
in dehydration, thirst and increased drinking
(polydypsia)[1]
. Insulin deficiency causes wasting
through increased breakdown and reduced
synthesis of proteins. Diabetes is a global epidemic
with an estimated worldwide prevalence of 246
million people in 2007 and forecasts to rise to 300
million by 2025.The number of people suffering
from diabetes all over the world has increased and
the disease now kills more people than AIDS[2]
.
Consequently, diabetes presents a major challenge
to healthcare systems around the world. Insulin and
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Pharmacology & Pharmacotherapeutics

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oral hypoglycemic agents, Sulfonylurea,
Biguanides and other a-Glucosidase inhibitors,
Thiazolidinediones are used to treat diabetes. The
all drugs are having the different mechanism of
action in disease but they may also produce a
number of undesirable effects, which include
frequent diarrhea, hypoglycemia, hepatotoxicity,
lactic acidosis, dyslipidemia, hypertension, and
hypercoagulability. Hence there is a need to search
for newer anti-diabetic agents that have therapeutic
efficacy with minimum side effects. Management
of diabetes without any side effect is still a
challenge to the medical community. There is
continuous search for alternative drugs. Therefore
it is prudent to look for options in natural
medicines for diabetes as well. Many traditional
plant and animal treatments for diabetes are also
used.
Inflammation is a complex biological response of
vascular tissues to harmful stimuli including
pathogens, irritants or damaged cells . It is a body
defense reaction to eliminate or limit the spread of
an injurious agent and is characterized by five
cardinal signs, redness (rubor), swelling (tumor),
heat (calor), pain (dolor) and loss of function
(function laesa)[3]
. The inflammatory process
involves a cascade of events elicited by numerous
stimuli that include infectious agents, ischemia,
antigen-antibody interaction and thermal or
physical injury. Non-steroidal anti-inflammatory
drugs (NSAIDs) are widely used in the treatment of
acute and chronic inflammation, pain and fever.
But the greatest disadvantage in presently available
synthetic drugs is that they cause gastrointestinal
irritation and reappearance of symptoms after
discontinuation. Therefore, there is a dire need for
screening and development of novel, but better
anti-inflammatory drugs and indigenous medicinal
products derived from the natural products could be
a logical source to find these[4]
.
Capra hircus belongs to family Bovidae. It is one of
the domesticated animal used for milk, meat and
fibre.These products are used therapeutically since
ancient time as medicine. Goats produce about 2%
of the world's total annual milk supply. The milk
generally averages 3.5% butterfat. Studies revealed
that goat milk has more beneficial properties to
health than cow milk. Goat’s milk has smaller fat
globules as well as higher levels of medium chain
fatty acids. This means that during digestion, each
fat globule and individual fatty acid will have a
larger surface-to-volume ratio resulting in a quicker
and easier digestion process[5]
.
Materials and Methods
Materials
The milk samples of domestic goat, Caprus hircus
were collected from intensively-farmed goat
population in the Medak region, Andhra Pradesh.
The milk was collected in the morning in sterile
screw bottles and kept on ice during transportation
to the laboratory where milk bottles were stored at
4°C. All reagents used in this study were of
analytical grade and obtained from SD Fine-Chem
Limited, Hyderabad, India.
Preparation of fatty acid extract of milk
from Caprus hircus
The fresh raw milk was collected from a healthy
Caprus hircus. The separating funnel and organic
solvent method are the chosen apparatus to extract
the fat of milk.The fatty acid is extracted using
organic solvent isopropanol[6][7]
. The Caprus hircus
milk Extract (CHME) obtained is evaluated for
qualitative identification of type of constituents
present.
Test Animals
Wistar albino rats (weighing 170+10g) of either sex
were used in this study. They were procured from
National Institute of Nutrition, Hyderabad. The
animals were kept in the standard polycarbonate
cages and provided with standard rat feed and
water ad libitum. The animals were housed under
standard environmental conditions with controlled
conditions of temperature (23 ± 2 ºC), humidity (50
± 20 %) and 12 hour light-dark cycles. The husk in
the cages was renewed twice a week to ensure
hygiene and maximum comfort for animals. The
animals were acclimatized for 5 days. The
experiments were performed according to the
current guidelines for the care of the laboratory
animals.
Determintation of acute toxicity (LD50)
The acute toxicity for hexane extract of Caprus
hircus milk was determined on both sex of wistar
rats, maintained under standard conditions. The
animals were fasted overnight prior to dosing.
Fixed dose method of Schedule Y given by

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DCGIwas adopted for toxicity studies. The milk
extract was administrated orally. The hexane
extract of Caprus hircus milk exhibiting no
mortality at 5000 mg/kg dose, was considered as
LD50 cut off dose (safe dose). So 1/50th
and 1/25th
of that were selected (100 and 200 mg/kg) for the
experiment as sub-maximal and maximal dose.
Ant diabetic activity
Streptozotocin induced diabetic rats were used for
the study of ant diabetic activity.
Induction of Diabetes to Experimental
Rats[8][9]
Diabetes was induced in the overnight fasted
animals by a single intra peritoneal injection of
freshly prepared solution of streptozotocin (STZ)
65 mg/kg body weight in 0.1M cold citrate buffer
pH 4.5. The animals were allowed to drink 5%
glucose solution to overcome the drug- induced
hypoglycemia. The control rats were injected with
citrate buffer alone as placebo. The animals were
considered diabetic if the blood glucose values
were >250 mg/dL on the third day after STZ
injection.
Experimental Design[10][11][12]
The rats were divided into five groups (n = 6).
Except group I which served as normal non-
diabetic control all other groups were comprised of
diabetic rats. Group II served as diabetic (STZ)
control. Groups III and IV, received CMHE (100
and 200 mg/kg and group V received standard
hypoglycemic drug Metformin (500 mg/kg b.w.,
p.o.) daily. The blood glucose levels was measured
just prior to and 2, 4 and 6h after drug
administration. Treatment is continued for 21
consecutive days. The fasting blood glucose levels
were estimated on days 0, 1, 7, 14 and 21 using
blood glucose test strips with elegance glucometer.
Antiinflammatory activity
Anti-inflammatory activity was evaluated by using
carrageenan induced paw edema model.
Carrageenan-induced paw edema
Albino rats of either sex weighing 150-200 grams
were divided into four groups of six animals each.
The dosage of the drugs administered to the
different groups was as follows. Group I -Control
(normal saline 0.5 ml/kg), Group - II and III –
CHME (100 mg/kg and 200 mg/kg, p.o.), Group
IV–reference standard antiinflammatory drug
(Diclofenac 10 mg/kg, p.o.). All the drugs were
administered orally[13]
. After one hour of the
administration of the drugs, 0.1 ml of 1% W/V
carrageenan solution in normal saline was injected
into the subplantar tissue of the left hind paw of the
rat and the right hind paw was served as the
control[14]
. The paw volume of the rats were
measured in the digital plethysmograph at the end
of 0 min, 30min, 60 min, 120 min, 180 min, 240
min, 300 min, 360 min[15]
.
Evaluation of antiinflammatory activity[16]
The percentage increase in paw edema of the
treated groups was compared with that of the
control and the inhibitory effect of the drugs was
studied. The relative potency of the drugs under
investigation was calculated based upon the
percentage inhibition of the inflammation.
Percentage inhibition was calculated using the
formula,
Percentageof inhibition of edema = (Vc-Vt)/Vc ×100
Where,
Vt = mean paw volume of test group
Vc = mean paw volume of control group.
Serum biochemical parameters
The blood was used for the estimation of serum
biochemical parameters viz. serum glutamate
oxaloacetate transaminase (SGOT), serum glutamate
pyruvate transaminase (SGPT), total cholesterol, Urea,
Creatinine were estimated by using commercially
available reagent. The blood was collected from retro-
orbital plexus of the overnight fasted rats under light
ether anesthesia and kept aside for ½h for clotting.
Serum is separated by centrifuging the sample at 6000
rpm for 20 min. The serum will be analyzed for total
protein, SGPT and SGOT levels[17][18]
.
Statistical Analysis
The experimental data were the blood sugar levels
as compared to the reference drug expressed as
mean ± standard error of mean (SEM). in a dose
dependant manner. Statistical significance was
analyzed by one-way analysis The fasting blood
glucose levels of normal, diabetic of variance
(ANOVA) followed byfallowed by LSD multiple
comparison test, where the P<0.05 was considered
significant.
Results and Discussion
The present study evaluated the amtihyperglycemic
activity of caprine milk fatty extract in
streptozotocin (STZ) induced rats. STZ induced

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diabetes rats have been widely used as a model for
evaluation of antidiabetic activity. STZ is taken up
by pancreatic β-cells via glucose transporter
GLUT2. The main cause of STZ-induced β -cell
death is alkylation of DNA by the nitrosourea
moiety of this compound. However, production of
NO(Nitric oxide) and reactive oxygen species may
also be involved in DNA fragmentation and other
deleterious effects of STZ In this model, diabetes
arises from irreversible destruction of the β-islet
cells of the pancreas, causing degranulation or
reduction of insulin secretion and insulin is
markedly depleted, but not absent. The isopropanol
extract of Caprus hircus produced significant
changes in the streptazotocin induced diabetic rats
by reducing the glucose levels considerably. The
prolonged treatment of caprine milk extract on
streptazotocin-induced diabetic rats produced
consistent reduction in the blood glucose levels.
There was significant decrease in blood glucose
level of CHME and Standard treated group with
mean values of normal control group. On the last
(21st) day, there was significant (p<0.0001)
decrease in blood glucose level of standard, CHME
treated groups with the mean values of
157.6±3.656**and 117.6±3.839** respectively as
compared with a mean value 317.2±4.244 of
diabetic control group. The blood glucose data
obtained clearly indicate that the Caprine milk
extract produced significant and consistent anti-
hyperglycemic effect in streptazotocin -induced
diabetic rats. The continuous treatment with
Caprine milk extract for a period of 21 days
produced a significant decrease in the blood
glucose levels of the diabetic rats, but not in the
normal rats. It is possible that the drug may be
acting by potentiating the pancreatic secreation or
increasing the glucose uptake. The results of effect
of isopropanol extract of Caprus hircus milk on
Blood Glucose level before treatment and after
treatment are summarised in Table-1.
Serum cholesterol, urea, creatinine, SGOT, SGPT
levels are decreased significantly by the
isopropanol extract of caprine milk when compared
with the diabetic control. The extract exhibited
anti-diabetic property in streptozotocin induced rats
as exhibited by the blood glucose levels.(Table 2)
The study of anti-inflammatory activity of
isopropanol extract of Caprus hircus against
carrageenan induced paw edema shows that the
extract has significant effect on inflammation and
markedly reduced the swelling. The percentage
reduction in the paw volume in the group of
animals treated with Caprine milk extract 100mg
was 56.08% and for the 200mg/kg was 71.73% at 4
hours. It shows that the plant extract have
significant (P <0.01) anti-inflammatory effect and
the results were compared with indomethacin
10mg/kg and show percentage paw volume
reduction of 66.30%. (Table 3). Carrageenan
induced hind paw edema is the standard
experimental model of acute inflammation.
Carrageenan is the phlogistic agent of choice for
testing anti-inflammatory drugs as it is not known
to be antigenic and is devoid of apparent systemic
effects. Moreover, the experimental model exhibits
a high degree of reproducibility. Carrageenan
induced edema is a biphasic response. The first
Phase is mediated through the release of histamine,
serotonin and kinins whereas the second phase is
related to the release of prostaglandin and slow
reacting substances with peak at 4h.
Table 1: Effect of Fatty extract of Caprus hircus milk on the blood glucose concentration in
streptazotocin induced rats
Groups Dose
mg/kg
Blood glucose concentration(mmolL-1
)
initial 1 d 7 d 14 d 21 d
Normal Control - 78.40±0.75 79.93±0.67 83.85±0.92 82.36±0.5693 80.38±0.562
Diabetic control
(Streptozotocin)
65 279.9±3.94 283.0±3.93 288.9±3.97 303.1±4.219 317.2±4.244
CHME I 100 286.0±3.28 280.8±2.17* 256.9±3.29* 223.4±1.944* 157.6±3.656**
CHME II 200 280.9±4.20 257.1±3.91* 219.9±3.28* 173.6±2.351** 117.6±3.839**
Standard (Metformin) 500 278.7±4.23* 259.5±4.77* 214.6±4.05** 165.1±4.096** 115.8±5.037**
P < 0.05 when compared to control. * P < 0.01. ** P < 0.001 when compared to control. Number of animal / in
each group = 6; Data expressed in mean ± SEM

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Table 2: Effect of Fatty extract of Caprus hircus milk on the serum biochemical parameters
Biochemical parameters vehicle Diabetic control standard (Metformin) Extract
(100 mg)
Extract
(200 mg)
Glucose(mg/dl) 81.43 ±2 .3 316.25 ± 2.6 206.59 ± 2.5 148.57 ±4.8 149.36 ±3.1
Cholesterol(mg/dl) 128 ± 3.5 224.5±6.70 140.26±4.46 185.25±4.55 154.67±3.60
Urea 22.0± 1.5 78.5± 3.47 30.16± 2.56 43.22± 2.0 32.15± 1.44
Creatinine (mg/dl) 0.76±0.08 1.72±0.16 0.95±0.10 1.22±0.05 1.04±0.06
SGPT 73.26±2.60 135.40±7.21 89.50±2.57 110.33±4.05 94.45±5.40
SGOT 26.45± 1.5 65.72± 3.84 37.55± 1.63 56.64± 2.30 48.96± 2.67
Total protein 7.8± 0.06 3.9± 0.02 8.4± 0.10 4.7± 0.05 6.5± 0.03
Values are expressed as mean ± SEM ( n = 6); * p < 0.001 compared with saline control
and ** p < 0.001 compared with STZ control group
Table 3: Effect of Fatty extract of Caprus hircus milk on paw edema induced by carrageenam in
rats:
Groups Dose
mg/k
g
Mean paw volume ± SEM(ml) and % Inhibition (P.I)
30 min 60 min 120 min 180 min 240 min 300 min 360 min
Normal
control
- 0.148±0.00
5
0.165±0.00
4
0.174±0.00
3
0.189±0.00
6
0.184±0.00
4
0.177±0.00
5
0.187±0.0
04
CHME I 100 0.147±0.00
4*
0.090±0.00
4 (45.45)
0.081±0.00
4* (53.44)
0.083±0.00
3 (56.08)
0.074±0.00
3* (58.82)
0.092±0.00
4 (48.02)
0.107±0.0
05
(42.78)
CHME II
200 0.142±0.00
5*
0.078±0.00
1* (52.72)
0.058±0.00
3** (66.66)
0.057±0.00
3** (69.84)
0.052±0.00
3*
(71.73)
0.082±0.00
3** (53.67)
0.093±0.0
05
(50.26)
Standard
(Diclofen
ac)
10 0.1292±0.0
04*
0.072±0.00
2** (56.36)
0.060±0.00
2** (65.51)
0.067±0.00
2* (64.55)
0.062±0.00
4 (66.30)
0.071±0.00
3** (59.88)
0.079±0.0
04
(57.75)
P < 0.05 when compared to control. * P < 0.01. when compared to control. Number of animal / in each
group = 6; Data expressed in mean ± SEM
Conclusion
It can be concluded that isopropanol extract of
Caprus hircus milk has anti-inflammatory activity
against carrageenan induced paw edema in rats and
a significant anti-hyperglycemic activity against
streptazotocin induced diabetic rats. The activity
may be due to their content of fatty acids like lauric
acid, palmiticacid, stearic acid, oliec acid. This
study demonstrates the efficacy of Caprus hircus
milk as an anti-hyperglycemic agent and anti-
inflammatory agent. It also scientifically justifies
the use of this milk extract as an anti-edematous
agent in folk medicine. However, further studies
are required to determine the constituents
responsible for its activities and further
authenticate its mechanism of action at the cellular
and molecular levels.
Conflict of Interests
The authors declare that they have no conflict of
interests.
Acknowledgment
We acknowledge to the management and the head
of the institute for making the facilities to carry out
the research work in a successful manner.
.

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