• Moath Kahtan Bashir Department of Pharmacology, College of Pharmacy, University of Mosul, Mosul, Iraq
  • Sabariah Ismail Centre for Drug Research, Universiti Sains Malaysia, 11800 Pulau Pinang, Malaysia
  • Ma H Q Department of Pharmaceutical Chemistry, School of Pharmacy, Universiti Sains Malaysia, 11800 Pulau Pinang, Malaysia
  • Nurul Huda Abdullah Centre for Drug Research, Universiti Sains Malaysia, 11800 Pulau Pinang, Malaysia
  • Abas Haji Hussin Centre for Drug Research, Universiti Sains Malaysia, 11800 Pulau Pinang, Malaysia



Objective: The aim of this study was to investigate the in vitro and ex vivo (acute and sub-chronic doses) effect of Phyllanthus niruri methanol extract (PNME) on the activity of cytosolic glutathione S-transferase (GST) in streptozotocin (STZ)-induced diabetic young female Sprague Dawley (SD) rats.

Methods: Young female SD rats were induced type I diabetes mellitus using STZ (60 mg/kg i.v.). The in vitro study was performed on cytosolic fraction of diabetic rat livers using PNME in concentrations of 0.01, 1 and 10 µg/ml. Meanwhile, ex vivo studies were performed on diabetic rats using PNME in doses of 500, 1000, 2000 and 5000 mg/kg p.o. for acute study (1 day) and 100, 500 and 2000 mg/kg/day p.o. for sub-chronic study (14 days). The GST activity in the cytosolic liver fraction was measured using spectrophotometric analysis.

Results: The in vitro study showed that, there is no significant effect of the three concentrations of PNME versus control on GST activity in cytosolic fraction of diabetic young female SD rat livers, while for ex vivo study, there is a significant dose independent induction effect on GST enzyme activity at all doses used in acute study. Meanwhile, sub-chronic study showed significant dose-independent induction effect at doses 500 and 2000 mg/kg.

Conclusion: This study suggests that Phyllanthus niruri may increase the clearance of drugs that are metabolized by GST enzyme in phase II metabolism when given concomitantly with this plant extract.

Key words: Phyllanthus niruri, GST, diabetic rats, cytosolic liver fraction, phase II metabolism 



1. Santos AR, Filho VC, Niero R, Viana AM, Moreno FN, Campos MM, Yunes RA, Calixto JB. Analgesic effects of callus culture extracts from selected species of Phyllanthus in mice. J. Pharm. Pharmacol. 1994;46:755-759.

2. De Souza TP, Holzschuh MH, Lionc MI, Ortega OGG, Petrovick PR. Validation of a LC method for the analysis of phenolic compounds from aqueous extract of Phyllanthus niruri aerial parts. J. Pharm. Biomed. Anal. 2002;30:351-356.

3. Nishiura JL, Campos EAH, Boim EAM, Heilberg IP, Schor EN. Phyllanthus niruri normalizes elevated urinary calcium levels in calcium stone forming (CSF) patients. Urol. Res. 2004;32:362-366.

4. Dharmaraj S, Jamaludin SA, Razak HM, Valliappan R, Ahmad NA, Harn GL, Ismail Z. The classification of Phyllanthus niruri Linn. according to location by infrared spectroscopy. Vib. Spectrosc. 2006;41:68-72.

5. Khanna AK, Rizvi F, Chander R. Lipid lowering activity of Phyllanthus niruri in hyperlipemic rats. J Ethnopharmacol. 2002;82:19-22.

6. Liu J, Lin H, Mcintosh H. Genus Phyllanthus for chronic hepatitis B virus infection: a systematic review. J. Viral. Hepat. 2001;8:358-366.

7. Naik AD, Juvekar AR. Effects of alkaloidal extract of Phyllanthus niruri on HIV replication. Indian J. Med. Sci. 2003;57:387-393.

8. Tsai CW, Yang JJ, Chen HW, Sheen LY, Lii CK. Garlic Organosulfur Compounds Upregulate the Expression of the {pi} Class of Glutathione S-Transferase in Rat Primary Hepatocytes. J. Nutr. 2005;135:2560-2565.

9. Gibson GG, Skett P. Introduction to drug metabolism, United Kingdum, Blackie Academic & Professional. 1994.

10. Bashir MK. Effect of Standardized Methanolic Extract of Phyllanthus Niruri (Linn.) on Phase II Drug Metabolism and its Molecular Mechanism Elucidation in Rat Liver. Unpublished doctoral dissertation, Universiti Sains Malaysia, Pulau Pinang, Malaysia. 2010.

11. Murugaiyah V, Chan KL. Determination of four lignans in Phyllanthus niruri L. by a simple high-performance liquid chromatography method with fluorescence detection. J. Chromatogr. A. 2007;1154:198-204.

12. Rooseboom M, Vermeulen NPE, Groot EDJ, Commandeur JNM. Tissue distribution of cytosolic B-elimination reactions of selenocysteine Se-conjugates in rat and human. Chem. Biol. Interact. 2002;140:243-264.

13. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem, 1976;72:248-254.

14. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-Transferase, The First Enzymatic Step In Mercapturic Acid Formation. J. Biol. Chem. 1974;249:7130-7139.

15. Han D, Hanawa N, Saberi B, Kaplowitz N. Mechanisms of Liver Injury. III. Role of glutathione redox status in liver injury. Am. J. Physiol. Gastrointest. Liver Physiol. 2006; 291:G1-G7.

16. Frova C. Glutathione transferases in the genomics era: New insights and perspectives. Biomol. eng. 2006;23:149-169.

17. Chin JH, Ismail S, Hussin AH. Induction of hepatic Glutathione S-Transferase activity by Orthosiphone stamineus benth. in STZ-induced diabetic rats. Malaysian Journal of Pharmaceutical Sciences, 2008;6:59-68.

18. Gao CM, Takezaki T, Wu JZ, Lic ZY, Liu YT, Li SP, Ding JH, Su P, Hu X, Xu TL, Sugimura H, Tajima K. Glutathione- S-transferases M1 (GSTM1) and GSTT1 genotype, smoking, consumption of alcohol and tea and risk of esophageal and stomach cancers: a case-control study of a high-incidence area in Jiangsu Province, China. Cancer Lett. 2002;188:95-102.

19. De Mattia G, Laurenti O, Bravi MC, Ghiselli A, Iuliano L, Balsano F. Effect of aldose reductase inhibition on glutathione redox status in erythrocytes of diabetic patients. Metabolism 1994;43:965-968.

20. Fortuno A, Jose GS, Moreno MU, Diez J, Zalba G. Oxidative stress and vascular remodeling. Exp. Physiol. 2005;90: 457-462.

21. Aragno M, Parola S, Tamagno E, Brignardello E, Manti R, Danni O, Boccuzzi G. Oxidative Derangement in Rat Synaptosomes Induced by Hyperglycaemia: Restorative Effect of Dehydroepiandrosterone Treatment. Biochem. Pharmacol. 2000;60:389-395.

22. Beisswenger PJ, Drummond KS, Nelson RG, Howell SK, Szwergold BS, Mauer M. Susceptibility to diabetic nephropathy is related to dicarbonyl and oxidative stress, Diabetes, 2005;54:3274-3281.

23. Konard RJ, Mikolanenko I, Tolar JF, Liu K, Kudlow JE. The potential mechanism of the diabetogenic action of streptozotocin : inhibition of pancreatic B-cell O-GlcNAc-selective N-acetyl-b-D-glucosaminidase. Biochem. J. 2001;356:31-41.

24. Agrawal R, Tyagi E, Shukla R, Nath C. A study of brain insulin receptors, AChE activity and oxidative stress in rat model of ICV STZ induced dementia. Neuropharmacology. 2009;56:779-787.

25. Iqbal M, Sharma SD, Okazaki Y, Fujisawa M, Okada1 S. Dietary Supplementation of Curcumin Enhances Antioxidant and Phase II Metabolizing Enzymes in ddY Male Mice: Possible Role in Protection against Chemical Carcinogenesis and Toxicity. Pharmacol.Toxicol. 2003;92:33-38.

26. Rajeshkumar NV, Joy KL, Girija K, Ramsewak RS, Nair MG, Ramadasan K. Antitumour and anticarcinogenic activity of Phyllanthus amarus extract. J. Ethnopharmacol. 2002;81:17-22.

27. Shamasundar KV, Singh B, Thakur RS, Hussain A, Kiso Y, Hikino H. Antihepatoprotective principles of Phyllanthus niruri herbs. J. Ethnopharmacol., 1985;14:41-44.

28. Kapur V, Pillai KK, Hussain SZ, Balani DK. Hepatoprotective activity of jigrine on liver damage caused by alcohol, carbon tetrachloride and paracetamol in rats. Indian J. Pharmacol. 1994;26:35-40.

29. De S, Shukla VJ, Ravishankar B, Bhavasar GC. A preliminary study on the hepatoprotective activity of methanol extract of Paederia foetida leaf. Fitoterapia, 1996;LX VII:106-109.

30. Emmanuel S, Amalaraj T, Ignacimuthu S. Effect of coumestans isolated from the leaves of Wedelia calandulaceae in paracetamol induced liver damage. Indian J. Exp. Biol. 2001;39:1305-1307.

31. Lee JS, Obach RS, Fisher MB. Drug-drug interactions and the cytochrome P450. In: Bachmann, KA, Ring BJ, Wrighton SA, eds. Drug Metabolizing Enzymes, Fontis Media S.A. and Marcel Dekker, Inc., New York, 2003.

32. Myhrstad MW, Carlsen H, Nordstr¨om O, Blomhoff R, Moskaug JØ. Flavonoids increase the intracellular glutathione level by transactivation of the glutamylcysteine synthetase catalytical subunit promoter. Free. Radic. Biol. Med. 2002;32:386-393.

33. Newairy ASA, Abdou HM. Protective role of flax lignans against lead acetate induced oxidative damage and hyperlipidemia in rats. Food.Chem. Toxicol. 2009;47:813-818.

34. Harish R, Shivanandappa T. Antioxidant activity and hepato-protective potential of Phyllanthus niruri. Food. Chem. 2006;95:180-185.

35. Sabir SM, Rocha JBT. Water-extractable phytochemicals from Phyllanthus niruri exhibit distinct in vitro antioxidant and in vivo hepatoprotective activity against paracetamol-induced liver damage in mice. Food.Chem. 2008;111:845-851.

36. Heim KE, Tagliaferro AR, Bobilya DJ. Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. J. Nutr. Biochem. 2002;13:572-584.

37. Bernini R, Gualandi G, Crestini C, Barontini M, Belfiore MC, Willför S, Eklund P, Saladino R. A novel and efficient synthesis of highly oxidized lignans by a methyltrioxorhenium/hydrogen peroxide catalytic system. Studies on their apoptogenic and antioxidant activity. Bioorg. Med. Chem, 2009;17:5676-5682.
268 Views | 310 Downloads
How to Cite
Bashir, M. K., S. Ismail, M. H. Q, N. H. Abdullah, and A. H. Hussin. “THE IN VITRO AND EX VIVO EFFECT OF PHYLLANTHUS NIRURI METHANOL EXTRACT ON HEPATIC GLUTATHIONE S-TRANSFERASE ACTIVITY IN STZ-INDUCED DIABETIC SPRAGUE DAWLEY RATS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 8, no. 5, Sept. 2015, pp. 156-9,
Original Article(s)