1. The effects of ginseng (Panax quinquefolius) treatment on chronic diabetic complications
Subhrojit Sen & Subrata Chakrabarti
Dept. of Pathology
University of Western Ontario

2. Introduction
Diabetes mellitus is a heterogeneous clinical syndrome in which the central feature is a chronic elevation of the blood glucose concentration.
It occurs due to deficiency of insulin (absolute) or resistance to insulin (relative).
40% diabetics develop long term complications affecting various organs including retina, heart and kidney.

3. Types of diabetes
Type I diabetes Results from the body's failure to produce insulin. It is estimated that 5-10% of Americans who are diagnosed with diabetes have type 1 diabetes.
Type II diabetes Results from insulin resistance (a condition in which the body fails to properly use insulin), combined with relative insulin deficiency. Most Americans who are diagnosed with diabetes have type 2 diabetes.
Gestational diabetes Occurring during pregnancy. Gestational diabetes affects about 4% of all pregnant women - about 135,000 cases in the United States each year.

4. Chronic diabetic complications Diabetic Cardiomyopathy
CHRONIC COMPLICATIONS OF DIABETES
Retinopathy
Nephropathy
Chronic diabetic complications
Diabetic Cardiomyopathy
Diabetic Neuropathy
Others

5. Progressive Background
Retinopathy
Progressive Background
i) Neovascularization i) increased capillary
permeability
ii) closure of retinal
capillaries
iii) vascular leakage
retinal edema
iv) accumulation of
lipids
v) retinal ischemia

6. Nephropathy:
Microalbuminuria
elevated blood pressure progresses to macroproteinuria
decline in renal function.

7. Diabetic Neuropathy:
Peripheral
Focal

8. Diabetic cardiomyopathy:
microvascular narrowing
micro aneurysm (increased risk of rupture)
hypertrophy
defects in collagen structure
myocardial fibrosis
perivascular fibrosis

9. Signalling pathways involved in diabetic complications
Hyperglycemia
Aldose reductase pathway
Protein kinase C
Oxidative stress
AGE/RAGE
Growth factors (TGF-β, VEGF etc.) ET
vasoconstriction
Permeability
PKC, MAPK
Blood flow
Growth Factors
Extracellular Matrix Protein Angiogenesis
Khan et al, Current Vascular Pharmacology, 2006, Vol 4, No. 1

10. Ginseng has been shown to have antioxidant effect
Ginseng has been shown to have antioxidant effect. Prevents oxidative lipid peroxidation and kidney damage in rats. Ginsenosides are the active components of ginseng found mainly in the roots. Structurally, ginsenosides are divided into two types: protopanaxadiol ginsenosides (PPDGs) and protopanaxatriol ginsenosides (PPTGS).
North American Ginseng

11. Objective
The present study will investigate the
effects of ginseng on glucose-induced changes in human vascular endothelial cells (HUVECs)
biochemical, functional and structural changes in heart, kidney and retina of insulin-dependent (IDDM) and noninsulin-dependent (NIDDM) mouse model.

12. Experiments on HUVECs
HUVECs were incubated with 25 mM glucose for 24 hrs. Aqueous and ethanolic extracts of ginseng were given at conc. of 5 mg/ml.
Glucose-induced
i) fibronectin (FN)
ii) FN protein level
iii) EDB+FN
iv) Vascular Endothelial Growth Factor (VEGF)
v) endothelin (ET-1) mRNA levels were measured.

13. Animal models used in the present study
Type 1 diabetes
STZ mice
Type 2 diabetes
db/db mice

14. Experimental Design Mice Type 1 STZ induction (IP, 65 mg/kg BW)
Type 2 db/db mice
Control
Diabetic DC
Diabetic +
Ginseng (DT)
(200mg/Kg BW for 2 months after onset of diabetes by oral gavage)
Control NC
Diabetic
db/db NC

15. Experimental parameters
Body weight
Heart weight
Biochemical parameters from
- blood :
a) Glucose
- urine
a) Albumin by ELISA
b) Creatinine by ALKALINE PICRATE METHOD

16. Preliminary results on HUVEC
FN-mRNA levels in Endothelial cells
0.5 1
1.5 2 NG HG
NGI
HGI
NGGE
HGGE
NGGA
HGGA
Groups
mRNA/18s RNA *
EDB+FN mRNA levels in endothelial cells
2.5
mRNA/18 sRNA
NG: 5mM glucose; HG: 25mM High glucose; I = insulin; GA: aqueous extract of ginseng; GE = ethanolic extract of ginseng. * = significantly different from other groups
ALCOHOLIC EXTRACTS APPEARED TO BE MORE EFFECTIVE

17. FN protein levels in endothelial cells* **
NG: 5mM glucose; HG: 25mM High glucose; I = insulin; GA: aqueous extract of ginseng; GE = ethanolic extract of ginseng. * = significantly different from other groups, * *= significantly different with HG and HGGA.
ALCOHOLIC EXTRACTS APPEARED TO BE MORE EFFECTIVE

18. * *
NG: 5mM glucose; HG: 25mM High glucose; I = insulin; GA: aqueous extract of ginseng; GE = ethanolic extract of ginseng. * = significantly different from other groups
ALCOHOLIC EXTRACTS APPEARED TO BE MORE EFFECTIVE

19. Preliminary results on diabetic mouse model
Groups
Body
wt.
(gms)
Heart
(mgm)
Fasting Blood
Glucose
mmol/L
Urinary
Albumin
(μg/ml)
Urinary Creatinine (mg/ml) NC
28±5.5
124.1±10.2
8.86±2.7
0.128±0.01
0.225±0.05 DC
24±1.6
121.12±2.5
27.4±5.5*
0.182±0.02*
0.350±0.025* DT
(ginseng
treated)
27.2±1.7
133.96±5.5
15.28±8.5
0.138±0.007
0.235±0.02
db/db
44.4±1.5* -
29.08±2.5*
0.184±0.02
0.312±0.01*
THE ABOVE PARAMETERS WERE MEASURED AFTER 2 MONTHS OF ONSET OF DIABETES IN CASE OF IDDM MODEL. THE PARAMETERS IN db/db MICE WERE MEASURED AT 12 WEEKS OF AGE (Duration of diabetes 4 weeks).

20. Conclusion
In HUVECs:
Glucose caused upregulation of FN, EDB+FN, VEGF and ET-1.
Treatment with both aqueous and ethanolic extract of ginseng (5mg/ml) caused diminution of the above parameters.
In STZ mice:
Treatment with ethanolic extract of ginseng (200mg/kg BW/day) for 2 months caused significant diminution of fasting blood glucose, urinary albumin and creatinine levels.
In db/db mice:
Enhancement of fasting blood glucose, urinary albumin and creatinine levels occurred after 12 weeks of age.

21. Acknowledgement
I sincerely acknowledge the guidance of my supervisor, Dr. Subrata Chakrabarti and Dr. Ed Lui, Dept. of Physiology & Pharmacology.
I also acknowledge the guidance and help provided by all members of my laboratory.
Shali Chen
2) Biao Feng
3) Jane Chiu
4) Kara McArthur
5) Yuexiu Wu
6) Chunyan Wang
7) Biju George