Presentation about Murraya Koenigi (Curry leaves) antidiabetic activity on Streptozotocin-Nicotinamide Rats model.

1. Antihyperglycemic Activity of Murraya koenigii
Leaves Extract on Blood Sugar Level in
Streptozotocin Nicotinamide Induced Diabetes
in Rats
Presented by Under the guidance of
Prashant K Sahu Dr. Navneet Khurana
M.Pharmacy (Pharmacology) Dr. Sazal Patyar
Reg. No. 12105615

3. Introduction
Rationale & Novelty of the Study
Materials and Methods
Results
Discussion
Conclusion
References
Contents

4. Introduction
 Diabetes (DM) is a chronic, metabolic disease characterized by elevated levels of
blood glucose (or blood sugar), which leads over time to serious damage to the heart,
blood vessels, eyes, kidneys and nerves.
 The two major pathophysiologies of DM are impairment of insulin action and
secretion.

5. Continue…
 Type 1 diabetes, once known as juvenile diabetes or insulin-dependent
diabetes, is a chronic condition in which the pancreas produces little or no
insulin by itself.
 The most common is type 2 diabetes, usually in adults, which occurs when the
body becomes resistant to insulin or doesn't make enough insulin.

6. Pathophysiology of Type 2 Diabetes

7. Signs and Symptoms
of Type 2 DM
 Type 2 diabetes often develop
slowly. In fact, person living with
type 2 diabetes for years and not
know it. When signs and
symptoms are present, they may
include:

8. Rationale & Novelty of the study
Conventional oral drugs used for DM have been proven for their characteristic adverse effects
like hypoglycemia and others. Therefore, the research has still attention to develop a novel
molecular drug with maximum effective and negligible to no side effects.
Murraya Koenigii leaves have been shown to have anti-diabetic effect in STZ induced type 1
diabetes in rats. However, the effects of this plant in STZ-NA (T2DM) caused diabetes in rats have
only been documented in a few trials, prompting researchers to look into the possibility of
generating a medicine with a similar mechanism of action as the plant extract this research work
was done.

9. Materials and Methods
Drug and chemicals:
Streptozotocin, Nicotinamide,
Metformin pure form powder,
and Methanol were procured
and all reagents and chemicals
used were of analytical grade
and stored in a refrigerator at
-4°C.
Collection and authentication
of plant material:
Leaves of Murraya koenigii
were collected and washed
thoroughly and later dried and
then powdered using a grinder.

10.  Metformin solution of conc. 20 mg/ml was prepared
and its dose was selected as 50 mg/kg body weight
per oral (bw, p.o).
 Methanolic and aqueous extracts of Murraya koenigii
leaves were prepared using Soxhalet apparatus until
dark brown colour of aqueous extract turned to pale
yellow while in case of methanolic extract, the
appearance of dark green to colourless.
Preparation of
Metformin
solution and
Murraya
koenigii leaves
extracts

11. Continue…
 Each extract solution was prepared by dissolving 400 mg of extract in 4 ml of
DW to attain a concentration of 100 mg/ml and its dose was selected as 200
mg/kg b w, p. o.
Acute toxicity study of extracts
The toxicity study was conducted as per OECD-423 guidelines. The dose of 2000
mg/kg b/w of AEMK and MEMK taken as a starting dose and were orally
administrated to healthy Wistar rats. Lethal dose 50% (LD50) was determined and
1/10th of LD50 was taken as therapeutic dose for antidiabetic activity.

12. Experimental Animals
 Healthy albino Wistar rats of either sex were obtained and was acclimatized
for 7 days.
 Experiment study was compiled with the guidelines of CPCSEA.
Inclusion Criteria Exclusion Criteria
Albino Wistar rats of either sex
weighing 120-250 gm.
Pregnant rats and those that have
delivered Once.
Albino Wistar rats with normal
behaviour and activity.
Albino Wistar rats that were
previously used for any other
experimental purpose.

13. STZ-NA induced diabetes in rats
 The study was performed for 28 days by repeated oral administration.
Diabetes was induced in overnight fasted rats of Group II to V was injected ip
with a single dose of NA (110 mg/kg b w) first, followed by a single dose of
STZ (55 mg/kg b w) once. After 6 hours, STZ-NA injected rats were fed orally
5% glucose solution for the next 24 h to prevent fatal hypoglycemia.
 After 72h of STZ-NA injection the blood sugar level of rats was measured for
confirmation of diabetes.

14. Experimental
Design
 STZ-NA diabetic (STZNAD) rats of
either sex were randomly divided
into 4 groups (six animals each
group) and one group consisting
6 ND rats and all animals had
free access to water and animal
diet. Oral administration of MET,
AEMK and MEMK were given at
once daily for 28 days.
Group 1
NC
Group 2
DC
Group 3
MET
Group 4
AEMK
Group 5
MEMK
ND rats
received
DW and
served
as
normal
control
STZNAD
rats
received
DW and
served as
diabetic
control
STZNAD rats
received
MET-
100
mg/kg/day,
p. o
STZNAD rats
received
AEMK-200
mg/kg/day,
p. o
STZNAD
rats
received
MEMK-200
mg/kg/day
, p. o

15. Monitoring of body weight and blood sugar level
during treatment
Body weights of rats were measured on (0, 14th and 28th
days). Blood samples of rats were collected from tails
and BSL was checked. BSL readings were taken at every
week interval of the study period (0, 7th, 14th, 21st and
28th days). All animals were sacrificed by cervical
decapitation at the end of study.
Statistical Analysis
Data were expressed as mean ± standard error means
(SEM). Data analysis was performed by using Graph Pad
Prism software. All data were analyzed by using two-way
repeated ANOVA and Tukey’s test was applied for Post Hoc
analysis. The value of p < 0.05 was considered to be
statistically significant.

16. Results
Acute toxicity study of leaves extracts
Study observed no mortality or any toxic reactions within 4 h and after 14 days
by oral administration of AEMK and MEMK even at the highest dose (2000 mg/kg)
and the dose of 200 mg/kg was considered safe at 1/10th dose of LD50.
Effect of leaves extracts on body weight
No significant changes in body weight (g) of rats were observed (Table 1).

17. Table 1
Effect of leaves
extracts on body
weight in STZ-NA
induced diabetic rats.
Group Day 0 (g) Day 14 (g) Day 28 (g)
I NC 201.50 ±
20.43
207.16 ±
20.34
213.16 ±
19.16
II DC 156.83 ±
08.27
171.83 ±
09.78
166.16 ±
11.64
III MET 138.83 ±
04.29
149.33 ±
03.85
146.83 ±
05.23
IV AEMK 187.00 ±
17.70
207.5 ±
21.73
172.6 ±
12.46
V MEMK 157.16 ±
12.03
174.33 ±
12.81
169.83 ±
12.54

18. Effect of leaves extracts on blood sugar level
Oral administration of AEMK (200 mg/ kg b w) and MEMK (200 mg/kg b w) caused
the significant reduction in BSL (mg/dL) as compared to the diabetic group.
(Table 2)

19. Table 2
Effect of leaves on
blood sugar level in
STZ-NA induced
diabetic rats
Group Day 0
(mg/dL)
Day 7
(mg/dL)
Day 14
(mg/dL)
Day 21
(mg/dL)
Day 28
(mg/dL)
I NC 065.00 ±
00.80
066.10 ±
01.20
064.6 ± 02.30 064.3 ± 02.04 066.5 ± 01.90
II DC 483.83 ±
50.57
450.00 ±
51.37
406.60 ±
22.46
393.50 ±
39.12
396.60 ±
36.22
III MET 323.30 ±
24.66*
168.50 ±
08.04***
133.60 ±
10.41***
153.50 ±
20.87***
140.00 ±
19.65***
IV AEMK 336.00 ±
37.21
195.00 ±
12.95***
161.10 ±
09.41***
173.30 ±
24.59***
154.10 ±
17.73***
V MEMK 380.16 ±
51.55
201.60 ±
04.99***
172.60 ±
33.06***
261.10 ±
41.26*
231.60 ±
39.66**
ANOVA F 16.215
P<0.0001
F 30.507
P<0.0001
F 46.816
P<0.0001
F 17.685
P<0.0001
F 21.999
P<0.0001

20. Discussion
 Several alkaloids present in the plant are reported as oral potential
hypoglycemic agents in-vivo and their effects on glucose uptake.
 The anti-diabetic effects of AEMK and MEMK in STZ-NA-induced diabetes in
rats were investigated in this work. In diabetic rats, continuous treatment
with AEMK and MEMK at the same dose of for 28 days resulted in a substantial
decrease in BSL (p 0.05).

21. Continue…
 Mahanimbine which is a major constituent of this plant has shown
hypoglycemic activities in different studies.
 Murraya koenigii defends pancreatic cells of Langerhans islets on the account
of flavonoids and phenolics as well as synergizes hypoglycemic activity due to
the major portion of naturally occurring carbazole alkaloids like koenidine
and mahanimbine.

22. Conclusion
 Murraya koenigii could lower BSL in diabetic rats so it is proven as an
effective, safe anti-diabetic agent.
 Though it is suggested that to evaluate the influence on the lipid profile and
antioxidant levels in diabetic rats by elaborating distinct activities of isolated
bioactives in the plant.

23.  https://www.who.int/health-topics/diabetes#tab=tab_1
 Paul S, Bandyopadhyay TK, Bhattacharyya A. Immunomodulatory effect of leaf extract of
Murraya koenigii in diabetic mice. Immunopharmacol Immunotoxicol 33(4):691- 9. (2011).
 J, Taneja I, et al. Naturally occurring carbazole alkaloids from Murraya koenigii as potential
antidiabetic agents. J Nat Prod. 79(5):1276-84. (2016).
 https://www.amjmed.com/article/S0002-9343%2813%2900485-3/fulltext
 OECD/OCDE. Acute Oral Toxicity–Acute Toxic Class Method. OECD Guidel Test Chem. 423:1-
14. (2001).
 https://iamdiabetic.in/article/type-2-diabetes-symptoms/
 Kumar BD, Krishnakumar K, Jaganathan SK, Mandal M. Effect of mangiferin and mahanimbine
on glucose utilization in 3T3-L1 cells. Pharmacogn Mag. 9(33):72-5. (2013).
 Phatak RS, Matule SM. Beneficial effects of Murraya koenigii leaves chloroform extract (MKCE)
on erythrocyte, thrombocyte and leukocyte indices in lead-intoxicated mice. Biomed
Pharmacol J. 9(3): 1035-40. (2016).
References