we isolated FR7 and FR8 from ficus racemosa which have a antidiabetic effect but no detailed study of hypoglycemic property found yet.

1. ANTIDIABETIC AND HYPOGLYCEMIC EFFECT OF ISOLATED
FLAVONOIDS FROM FICUS RACEMOSA STEM BARK IN
STREPTOZOTOCIN DIABETIC RATS
DEPARTMENT OF PHARMACEUTICAL SCIENCES
BABA SAHEB BHIMRAO AMBEDKAR UNIVERSITY
LUCKNOW, U.P.
SESSION 2014-2015
A thesis presentation submitted
to
Babasaheb Bhimrao Ambedkar University
in partial fulfillment of the requirements for the degree
of
MASTER OF PHARMACY
In
pharmacology
SUPERVISOR
Dr. Sudipta Saha
Assistant professor
PRESENTED BY
Ghanendra
Roll No- 32006

2. INTRODUCTION
 Diabetes is a global epidemic with an estimated worldwide prevalence of
246 million people in 2007 and forecasts to rise to 300 million by 20251.
 Diabetes is a metabolic disorder.
 It characterized by chronic hyperglycaemia with disturbance of
carbohydrate, fat, and protein metabolism resulting from defects in insulin
secretion, insulin action or both.
 There are two main types of diabetes -
I. IDDM (insulin-dependent diabetes mellitus) - It results from the
body’s failure to produce insulin
II. NIDDM (non-insulin-dependent diabetes mellitus)- It occurs due
to insulin resistance, a condition in which cells fail to use insulin
properly

3. CONT...
 There are symptom of diabetes –
 Blurred vision, unusual thirst, Polydepsia, Polyurea
Rapid weight loss, Erectile dysfunction
 It is diagnosis by the several parameters –
 Fasting plasma glucose level ≥ 7.0 mmol/l (126 mg/dl).
 Plasma glucose ≥ 11.1 mmol/l (200 mg/dl) two hours after a
75 g oral glucose load as in a glucose tolerance test.
 Glycosylated hemoglobin (Hb A1C) ≥ 6.5%.

4. CONT...
 Treatment -
 In type 1 diabetes require insulin.
 In type 2 diabetes treatment will vary depend on blood sugar level.
 Prevention –
 Good nutrients for normal body weight
 sensible exercise
 Test sugar level

5. LITERATURE REVIEW
 Eleazu et al (2013) reported that streptozotocin (STZ) produced
cytotoxicity through DNA methylation, nitric oxide production, free radical
generation and alteration NF- κB (nuclear factor kappa-light-chain-enhancer
of activated B cells) cell signal transduction pathway.
 Arunachalam K et al (2013) stated that ficus amplissima smith: bark
extract improved the diabetic condition in STZ induced diabetic rats and
significantly enhanced the level of antioxidant enzyme.
 Peibo Li et al (2013) reported that the 13-week subchronic oral toxicity
study, daily doses of 50, 250, and 1250 mg/kg of naringin were well
tolerated and did not cause either lethality or toxic clinical symptoms and
changes in both sexes of rats except for slight body weight decrease.

6. CONT...
 Cheng D et al (2012) repoted that Ginkgo biloba extract (GBE) possesses
antihyperglycemic, antioxidant, and antihyperlipidemic activities in STZ-
induced chronic diabetic rats, which promisingly support the use of GBE as
a food supplement or an adjunct treatment for diabetics.
 Mahmoud M A et al (2012) stated that antihyperglycemic efficacy of the
citrus flavonoids, hesperidin and naringin, may be mediated via up-
regulating adipose tissue PPARγ and adiponectin in conjunction with down-
regulating adipose tissue resistin.
 Rahigude A et al (2012) stated that both narigenin and pioglitazone showed
improvement in cognitive behavior against diabetes induce memory
dysfunction via depleting elevated lipid peroxide and nitric oxide and
inhibition of elevated ChE activity.

7. RESEARCH ENVISAGED
previous report suggested that
flavonoids produced antidiabetic action
in streptozotocin induced diabetic rats.
No detail study had been performed
regarding their hypolipidemic and
toxicological action.
The overall objectives of this project –
Toxicological, hypoglycemic and
molecular docking study via PPARγ
and GLUT1 receptor

8. PLAN OF WORK
Drugs: FR7, FR8
and glibenclamide
Toxicant:
streptozotocin
(STZ)
Animal
study
(n=5)
Biochemical
estimation

9. CONT...
S.NO GROUP DOSE ROUTE OF
ADMINISTRATION
1 Normal
control
0.25mg/kg, CMC Oral
2 Diabetic
control
50mg/kg, STZ I.P
3 D + N 10mg/kg,
Glibenclamide
Oral
4 D + FR7 50mg/kg, FR7 Oral
5 D + FR8 50mg/kg, FR8 Oral

10. CONT...
Biochemical
estimation
Glucose level and
weight variation
on 1st 3rd 5th &7th
day
1. Oxidative
parameter:
SOD, CAT,
GSH, TBARS,
PC
Lipid profile:
HDL LDL,
TC, TG &
VLDL
SGOT &
SGPT
level in
plasma
Histopathology
and docking
study
Statistical
analysis:
ANOVA

11. RESULTS AND DISCUSSION
 Antidiabetic effect of isolated flavonoids:
 Diabetic rats treated with FR7 (Quercetin), FR8
(Naringenin) & G (glibenclamide) showed reduction of
glucose level in comparison to D (diabetic control).
0
100
200
300
400
N Control D Control D + G FR7 FR8
mg/dL
7 day
1 day
3 day
5 day
7 day
Absorpti-
on of
glucose
from
intestine
Neogluc-
ogenesis
Increase
glucose
level in
diabetic
rats

12. RESULTS AND DISCUSSION
 Effect of isolated flavonoids on body weight:
 D group showed the reduction in body weight in
STZ(streptozotocin) diabetic rats. Orally administration
of FR7 and FR8 displayed improvement in body weight
and treated with G showed increased in body weight.
0
50
100
150
200
N Control D control D + G FR7 FR8
Changeinbodywt.
7 day
1 day
3 day
5 day
7 day

13. RESULTS AND DISCUSSION
 Effect of flavonoids on glycogen content in liver:
 Glycogen content was massively decreased in D control
than N control.
 Again, we found that glycogen content was improved by
FR7 and FR8 treatment.
0
10
20
30
40
50
60
N Control D Control D + G FR7 FR8
mg/g
7 day
glycogen

14. RESULTS AND DISCUSSION
 Effect flavonoids lipid profile in plasma:
 We observed that TC, TG, LDL, VLDL levels were
increased and HDL level was deceased in toxic control
rats (D control).
 lipid profile improved via FR7, FR8 and G treatment.
0
50
100
150
200
N Contol D
Control
D + G FR7 FR8
mg/dL
7 day
TC
TG
HDL
LDL
VLDL

15. RESULT AND DISCUSSION
 Oxidative parameter in pancreas:
 We observed that GSH, SOD & CAT level were
decrease in D control than N control. The level of GSH,
SOD & CAT were restored via 7 day treatment therapy
of FR7 and FR8.
0
20
40
60
80
100
120
N Control D Control D + G FR7 FR8
SOD
CAT
GSH

16. RESULTS AND DISCUSSION
 Oxidative parameter in liver
 According to observation, the formation of TBARS and
PC were higher in D control as compared to N control.
Both formations were decreased when flavonoids
treated to STZ rats.
0
20
40
60
80
100
N Control D Control D + G FR7 FR8
TBARS
PC

17. RESULTS AND DISCUSSION
 CAT is mainly responsible for the catalytic
decomposition of H2O2 to oxygen and water.
 SOD is responsible for catalytic dismutation of free
radicals and decrease superoxide level.
 Reduced GSH converted to disulfide glutathione
during oxidative damage.
 TBARS and PC level reduced by inhibition of lipid
peroxidation protein carbonylation.

18. RESULTS AND DISCUSSION
 Effect of FR-7 & FR-8 on AST and ALT level:
 Isolated flavonoids also showed protective action on AST,
ALT levels in plasma. Serum enzyme levels were increased
during STZ treatment which was normalized during FR7 and
FR8 administration .
0
50
100
150
200
N Control D Control D + G FR7 FR8
u/dL
7day
AST
ALT

19. RESULTS AND DISCUSSION
 Histopathology of pancreatic tissue
N Control D Control
D + N FR7 FR8

20. RESULTS AND DISCUSSION
GLUT
1
1SUK:
TYR28,GLY31,
ARG126, TRP412
FR7
PDB
Target
Exothermic
reaction
Hydrogen bond formation
And interaction energy (-
6.2)
Docking study of FR7, FR8 on GLUT1 & PPARγ

21. RESULTS AND DISCUSSION
FR7
Target
PDB
Exothermic
reaction
Hydrogen bond formation
And interaction energy(-9.6)
Docking study ...

22. RESULTS AND DISCUSSION
FR8
GLUT1
GLU261, ALA405,
VAL406, PHE81,
SER80, MET77,
Exothermic
reaction
Hydrogen bond formation
And interaction energy(-5.9)
Docking study ...

23. RESULTS AND DISCUSSION
VAL248, PHE347, LEU237
PPARγ
Exothermic
reaction
Hydrogen bond formation And
interaction energy(-6.2)
Docking study ...
FR8

24. SUMMARY AND CONCLUSION
Whole plant have antidiabetic
property
Previous literature suggested that
flavonoid has antidibetic activity
We isolated four flavonoids from
whole plant and performed
antidiabetic activity of two compound
– FR7 and FR8

25. REFERENCES
 A report of WHO named diabetes.
 Sapiro M, Welss JP. Diabetes insipidus: A review. Diabetes and metab.2012;
S8:001.
 Cheng D, Liang B, Li Y. Antihyperglycemic effect of Ginkgo biloba extract in
Streptozotocin - induced diabetes in rats. BioMed research international.2012;
2013.
 Chukwuma S, Essien NU. Review of the mechanism of cell death resulting from
streptozotocin challenge in experimental animals, its practical use and potential
risk to humans. Journal of diabetes & metabolic disorders. 2013; 12:60.
 Mahmoud AM, Ahmed OM, Monein AA, Ashour MB. Upregulation of PPARγ
mediates the antidiabetic effects of flavonoids in type 2 diabetic rats.
International journal of bioassays. 2012; 02(05): 756- 761.

26. CONT...
 Kushwaha PS, Raj V, Singh AK, Keshari AK, Saraf SA, Mandal SC, et al.
Antidiabetic effects of isolated sterols from Ficus racemosa leaves. RSC
Advances. 2015;5(44):35230-7.
 Yadav RK, Keshari AK, Kaithwas G, Maity S, Saha S. Antidiabetic and
hypolipidemic effect of Ficus racemosa petroleum ether extract in streptozotocin
induced diabetic Albino Rats. BioMed Research International. 2014
 Saha S, Chan DSZ, Lee CY, Wong W, New LS, Chui WK, et al.
Pyrrolidinediones reduce the toxicity of thiazolidinediones and modify their anti-
diabetic and anti-cancer properties. European journal of pharmacology.
2012;697(1):13-23.
 Sophia D, Manoharan S. Hypolipidemic activities of Ficus racemosa Linn. bark
in alloxan induced diabetic rats. African Journal of Traditional, Complementary
and Alternative Medicines. 2008;4(3):279-88.
 Szkudelski T. The mechanism of alloxan and streptozotocin action in B cells of
the rat pancreas. Physiological research. 2001;50(6):537-46.