Antioxidant role of mulberry (Morus indica L. cv. Anantha) leaves in streptozotocin-diabetic rats

1. Antioxidant role of mulberry (Morus indica L. cv. Anantha)leaves in streptozotocin-diabetic rats • Bondada Andallua *, N.Ch. Varadacharyulub • a Department of Home Science, Sri Sathya Sai Institute of Higher Learning, Anantapur-515001, India • b Department of Biochemistry, Sri Krishnadevaraya University, India 系別：生科四甲 學號：91390391 姓名：吳佳煒 Received 20 December 2002 received in revised form 7 July 2003; accepted 7 July 2003

2. Abstract • The antihyperglycemic and antioxidant role of mulberry (Morus indica L.) leaves were investigated. Model：Wistar rats • Standard diet • 25% dry mulberry leaf powder and standard diet For 8 weeks

3. Abstract • Key point： antihyperglycemic and antioxidant role of mulberry effect： • on blood glucose • lipid peroxidation • reduced glutathione (GSH) concentrations and on the activity of glucose-6-phosphate dehydrogenase (G6PDH) • various antioxidant enzymes in erythrocytes

4. Abstract Results： • lipid peroxidation • fasting blood glucose concentrations • glutathione GSH concentrations • activity of glucose-6-phosphate dehydrogenase (G6PDH) • antioxidant enzymes

5. Introduction • Hyperglycemia 血糖過高症 • Free radicals result in the consumption of antioxidant defenses may lead to： • disruption of cellular functions • oxidative damage to membranes • enhance susceptibility to lipid peroxidation.

6. Introduction • Total antioxidant status (TAS) of serum is reduced in type I diabetes • The reactive oxygen species are modulated by antioxidant enzymes： • Glutathione reductase (GR) • glutathione peroxidase (GPx) • glutathione-S-transferase (GST) • superoxide dismutase(SOD) • catalase (CAT) • non-enzymatic scavengers like reduced glutathione.

7. Introduction • Mulberry (M. alba L.) leaves containing many nutritional components： • Flavones 黃酮 • Steroids 類固醇 • Triterpenes 三烯 • amino acids 氨基酸 • Vitamins 維生素 • Moracin 桑辛素 • other trace minerals 無機物

8. Materials Animals • Male rats of Wistar strain from Hyderabad • weight range 150~200g • air-conditioned room (25° ±1℃) • light from 7 a.m. to 7 p.m. • Divide into 4 groups

9. Materials Wistar rats • I： normal control • II：normal treated with mulberry leaf powder • III：diabetic control • IV：diabetic treated with mulberry leaf powder

10. Materials Leaves • The mulberry leaves (M. indica L. cv. Anantha) • The 4th and the 5th leaves from the apex of healthy plants were plucked • washed thoroughly under running tap water • shade dried for 3 days and ground to a fine powder in an electric mixer

11. Methods • Groups III and IV (diabetic) • by a single i.p. injection of streptozotocin(STZ) (55 mg/kg) prepared in 1 ml of freshly prepared 0.1 mol/l citrate buffer after an overnight fast. • Groups I and II (Control) were injected with citrate buffer alone. • Streptozotocin injected animals were given 5% glucose water for 24 h following streptozotocin injection to prevent initial drug-induced hypoglycemic mortality

12. Methods streptozotocin • 注射streptozotocin，以破壞分泌胰島素的β胰島細胞，進而形成高血糖的情形 。 • 增加大鼠之進食量、飲水量及排泄量，並使體重變輕、血糖升高。 • 人類糖尿病的症狀：吃多、喝多、尿多

13. Methods - Biochemical estimations • Erythrocytes were separated from whole blood • Erythrocytes were washed thrice with 0.9% NaCl • suspended in 1 volume of 0.9% NaCl • The packed cell volume was adjusted to 5% with PBS (pH 7.4)

14. Student's T test

15. Results & Discussion

16. Fasting Blood Glucose

17. Fasting blood glucose

18. decreased food consumption insignificantly

19. increased the body weight in diabetic rats

20. Mulberry • decreased food consumption insignificantly • increased the body weight in diabetic rats

23. fagomine which potentiates the glucose induced insulin release similar to the action of glibenclamide a sulfonylurea drug which triggers the release of endogenous insulin • increase in tissue uptake of glucose by mulberry leaves 桑葉生物鹼fagomine可顯著增加葡萄糖引起的胰島素含量增加，從而降低糖尿病小鼠血糖。

25. The effect of mulberry in controlling hyperglycemia and glycosuria could be due to: • the N-containing sugars isolated form mulberry leaves which inhibit the functions of • α-glucosidase • α-mannosidase • β-galactosidase

26. diabetic control rats vs. normal controls • glucose-6-phosphate dehydrogenase 13% • glutathione reductase 34% • glutathione peroxidase 20% • glutathione-S-transferase 30% • superoxide dismutase 46% • Catalase 59% p < 0.01 CATALASE可氧化分解身體內過量的尿酸.將H2O2還原成H2O的反應 Glutathione peroxidase:催化以glutathione為還原劑，將H2O2還原成H2O的反應

27. Diabetic control v.s mulberry • The activity of glucose-6-phosphate dehydrogenase (69%; p < 0.01) • glutathione reductase (95%; p < 0.01), glutathione peroxidase (151%; p < 0.01) • glutathione-S-transferase (24%; p < 0.01) • superoxide dismutase(106%; p < 0.01) • activity of Catalase 33% all p < 0.01

28. Conclusion • Mulberry leaves are palatable and effective in controlling hyperglycemia and glycosuria in STZ-diabetic rats. • The leaves exerted a rapid protective effect against lipid peroxidation by scavenging O2 and elevating the activities of antioxidant enzymes by virtue of antioxidant flavonoids and moracins present in the leaves. 桑辛素Moracin 這類化合物顯示了良好的抗菌和抗真菌作用，是植物抗真菌抗生素。 類黃酮 flavonoid一類不含氮的生物色素，包括花色素&FAA1;和花黃色素。類黃酮在植物中存在普遍，但在動物中出現較少，具有抗癌的作用。

29. Question & Answer Thanks your listening