1. Non-insulin dependent hypoglycemic effect of Gynostemma pentaphyllum water extract in streptozotocin induced non-insulin dependent diabetic rats
Pei-Hsiu Hung

2. Pei-Hsiu Hung 1,2, Jane-Yii Wu 2 and Shih-Liang Chang 3
1. Director of Traditional Chinese Medicin, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Taiwan
2. Department of BioIndustry Technology, Da-Yeh University, Taiwan
3. Director of Department of Medicinal Botanicals and Health Appications, Da-Yeh University, Taiwan

3. Introduction: Gynostemma pentaphyllum
an herbal medicinal plant known in Taiwan as ‘Jiao-Gu-Lan’ or ‘q-iye-dan’ .
Fiveleaf Gynostemma

4. Anti-tumor
Gypenoside induces apoptosis in human Hep3B and HA22T tumor cells
Chen et al., 1999, Cytobios Volume 100, Issue 393 , 1999, Pages 37-48
Regulation of Bcl-2 family molecules and activation of caspase cascade involved in gypenosides-induced apoptosis in human hepatoma cells
Wang et al.,Cancer Letters 183 (2002) 169–178
Chiu et al., 2003

5. Anti-oxidation
The Direct Release of Nitric Oxide by Gypenosides Derived from the Herb Gynostemma pentaphyllum
Tanner et al., NITRIC OXIDE
Vol. 3, No. 5, pp. 359–365 (1999)
Gypenosides derived from Gynostemma pentaphyllum suppress NO synthesis in murine macrophages by inhibiting iNOS enzymatic activity and attenuating NF-jB-mediated iNOS protein expression
Aktan et al., Nitric Oxide 8 (2003) 235–242

6. Anti-Hyperlipidemic
Phytopreventative Anti-Hyperlipidemic Effects Of Gynostemma Pentaphyllum In Rats
Megalli et al.,2005

7. Hypoglycemic activity
Natural medicines used in the traditional Chinese medical system for therapy of diabetes mellitus.
Li et al.
J of Enthopharmacology 2004, 92: 1-21.
A novel insulin-releasing substance, phanoside, from the plant Gynostemma pentaphyllum.
Norberg et al.
J. Biol. Chem., 2004,279(40):
1.文中提到70多種有調節血糖的中草藥，其中包括絞股藍。
2. 由絞股藍，以乙醇和正丁醇萃取物，再分離出一種分子量為914.5 Da的物質，命名為phanoside，有刺激胰島細胞分泌，以及改善血糖耐受性功能。

8. Purpose
Gynostemma pentaphyllum (GP) had been used in the treatment of Diabetes Miletus (DM), but the mechanism is still not clear enough.
The purpose of this study is to investigate the hypoglycemic effect in the water extract of GP, and to study the secretion of insulin and improvement of insulin resistance.

9. Method:water extract of GP
Gynostemma Pentaphyllum
add H2O，soaked for 30‘
add H2O，boiling for 1h
filtrate
Filtrate
Residue
add H2O，boiling for 1h
filtrate
Filtrate
Residue
Combined filtrate，Concentrated by vaccum，Cold storage for usage

10. Method：Animal Model
Male Wistar rats that weighed 300 – 350 g, ranging in age from 8 to 10 weeks,

11. Method：Animal Model STZ-induced IDDM rats
were produced by intravenous injection (i.v.) of 60 mg/kg streptozotocin (STZ) (Sigma Company, USA) after the rats had been fasting for 3 days.
Seven days after being treated with STZ, the rats' blood glucose levels were checked.
If the blood glucose level was greater than 350 mg/dl, the rat was included in the STZ-induced IDDM model

12. Method：Animal Model STZ-induced NIDDM rats
were produced by Intraperitoneal injection (i.p.) of 60 mg/kg streptozotocin (STZ) (Sigma Company, USA) after the rats had born for 3 days.
Blood glucose levels were determined ranging from 8 to 10 weeks after streptozotocin injection
the blood glucose level was ranging from 130 to 170 mg/dl

13. Method
Normal rats were applied to evaluate the lowering plasma glucose effect, and find the most effective dose and time.
Blood samples were taken from the femoral vein GP
30 min
60 min
Glucose
level
Normal rat→ 0, 50, 500 mg/kg

14. Method After finding out the most effective dose and time,
insulin level was assayed for evaluating the insulin secretion. GP
Glucose
Insulin
HOMA-IR
IDDM rats
NIDDM rats
X mg/kg
Y min

15. Homeostasis Model Assessment of Insulin Resistance (HOMA-IR)
Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) was also applied for evaluating the improvement of insulin sensitivity.
HOMA-IR = FI x FPG / 22.5
FI = fasting plasma insulin level (U/ml)
FPG = fasting plasma glucose level (mmol/l)
(Matthews, et al, 1985)

16. Statistical Analysis Mean±SEM Wilcoxon Signed Ranks Test
self-paired t-test
P<0.05

17. Result 1:
Three dosages, 0, 50, 500 mg/kg water extract of GP were applied orally for 30 minutes to test the optimal dose of hypoglycemic effect.
The most hypoglycemic effect was the dosage, 500 mg/kg.
18.44±3.50%

18. Glucose Lowering Effect
mg/kg 50
500
Befer
treatment
126.81
±6.69
129.02
±4.18
136.67
±5.37
After
30 min
110.25
112.33
±8.80
106.47
±4.12
HGA (mg/dl)
16.45
±5.48
16.71
±9.76
24.26
±5.04
HGA (%)
12.91
12.52
±6.54
18.44
±3.50 ＊ ＊
HGA: Hypoglycemic Activity

19. The effective dose of GP on the normal wistar rats＊ ＊

20. Result 2:
Three different animal models, normal, non-insulin dependent DM (NIDDM) and insulin dependent DM (IDDM) rats took 500 mg/kg water extract of GP orally to compare the hypoglycemic effects.
There were significant hypoglycemic effects of normal and NIDDM rats, but no significant hypoglycemic effects of IDDM rats.

21. The impact of GP on the plasma glucose between different rat models
GP 500 mg/kg
Before
treatment
After
30 min
GLE%
Control group (n=7)
130.67±4.98
106.47±3.82
-18.44
NIDDM group (n=10)
151.43±20.29
126.68±25.28
-16.51
IDDM group (n=8)
413.29±38.44
388.35±34.39
-5.93 ＊ ＊
GP: Gynostemma pentaphyllum water extract
GLE: Glucose Lowering Effect

22. Result 3:
In the assay of insulin, there were no significant change of insulin levels in NIDDM rats.
Plasma
Glucose ↓
Insulin
Secretion
Resistance ？ ？

23. The impact of GP on the plasma insulin of NIDDM-rat model
　Group
Insulin
(μU/ml)
before
after
Control (n=8)
0.828
±0.404
0.954
±0.496
NIDDM (n=8)
1.024
±0.630
0.79
±0.564

24. ？ X Result 4: The NIDDM rats were compared the change of HOMA-IR,
there were significant change of HOMA-IR in NIDDM rats.
Plasma
Glucose ↓
Secretion
Insulin
Resistance ？ X

25. The effect of GP on the HOMA-IR of NIDDM-rat model﹡

26. Conclusion:
The water extract of GP has the hypoglycemic effect in normal and NIDDM rats.
The mainly mechanism was a non-insulin dependent lowering plasma glucose manner through to improve the insulin resistance in STZ induced NIDDM rats.
Plasma
Glucose ↓
Secretion
Insulin
Resistance X V