Rosmarinic Acid Ameliorates Diabetic Nephropathy in Uninephrectomized Diabetic Rats

Document Type: Original Article


1 Department of Anatomy, Faculty of Medicine, Lorestan University of Medical Sciences and Razi Herbal Researches Center, Lorestan University of Medical Sciences, Khoram Abad, Iran

2 Department of Biochemistry, Faculty of Medicine, Lorestan University of Medical Sciences, Khoram Abad, Iran

3 Department of Anatomy, Faculty of Medicine, Lorestan University of Medical Sciences, Khoram Abad, Iran


Oxidative stress plays an important role in diabetic nephropathy pathogenesis. Rosmarinic acid, a plant phenolic compound, was first used as an antioxidant agent for inhibition of diabetic nephropathy.
Material and Methods
Forty male rats were uninephrectomized from the left flank.The rats were divided in four groups randomly; group one as control, group two diabetic untreatment, groups three and four treatment with rosmarinic acid by 100 or 200 mg/kg/d orally respectively. Diabetes was induced in the second, third and fourth groups by alloxan injection subcutaneously. After 8 weeks treatment, serum malondialdehyde was measured by thiobarbituric acid (TBA) test. Serum creatinine and serum urea were measured by kits. Kidney paraffin sections were prepared and stained by periodic acid Schiff method.Glomerular volume and glomerular number were estimated by stereological rules and glomerular sclerosis was studied semi-quantitatively. Data were analyzed by non-parametric Man Whitney test ( using SPSS 13 software) and P< 0.05 was considered significant.
Rosmarinic acid (100 or 200 mg/kg) significantly inhibited glomerular hypertrophy, glomerular number loss, glomerulosclerosis, lipid peroxidation, serum urea and creatinine compared with the diabetic untreated group. The level of glomerular number and serum malondialdehyde in the treated groups (100 mg/kg or 200 mg/kg of rosmarinic acid) was maintained at the same level as compared to the control group.
Rosmarinic   acid   could   significantly   reduce   glomerular   hypertrophy,   loss   of   glomerular   number, glomerulosclerosis and attenuated serum urea and serum creatinine in diabetic rats.


1. Vasavada N, Agarwal R. Role of oxidative stress in diabetic nephropathy. Adv Chronic Kidney Dis 2005; 12:146- 154.

2.  Schena FP, Gesualdo L. Pathogenetic mechanisms of diabetic nephropathy. J Am Nephrol 2005; 16:s30-s33.

3. Rodrigo R, Bosco C. Oxidative stress and protective effects of polyphenols: Comparative studies in human and rodent kidney. A review Comparative Biochemistry and Physiology 2006; 142 :317–327.

4. Sepici  DA, Açikgöz  S, Cevik  C, Sengelen  M, Yeşilada  E. Effects of in vivo antioxidant enzyme activities of myrtle oil in normoglycaemic and alloxan diabetic rabbits. J Ethnopharmacol 2007; 110:498–503.

5. Ha H, Hwang IA, Park JH, Lee HB. Role of reactive oxygen species in the pathogenesis of diabetic nephropathy. Diab Res Clin Prac 2008; 82:s42-s45.

6. Choi SW, Benzie IF, Ma SW, Strain JJ, Hannigan BM. Acute hyperglycemia and oxidative stress: Direct cause and effect? Free Radical Biology & medicine  2008 ;44:1217-1231.

7. Stephens JW, Khanolkar MP, Bain SC. The biological relevance and measurement of plasma markers of oxidative stress in diabetes and cardiovascular disease. Athersclerosis  2009; 202:321-329.

8. Allen DA, Harwood S, Varagunam M, Raftery MJ, Yaqoob MM. High glucose-induced oxidative stress causes apoptosis in proximal tubular epithelial cells and is mediated by multiple caspases. FASEB J 2003; 17:908-910.

9. Qattan KA, Thomson M, Muslim A.Garlic (Allium sativum) and ginger (Zingiber officinale)  attenuate structural nephropathy progression in streptozotocin-induced diabetic rats. e-SPEN, the European e-J Clin Nutr Metabol 2008; 3:e62-e71.

10. Huang Ss, Zheng Rl. Rosmarinic acid inhibits angiogenesis and its mechanism of action in vitro. Cancer Lett 2006; 239:271-280 .

11. Bakirel T, Bakirel U, Keleş OU, Ulgen SG, Yardibi H. In vivo assessment of antidiabetic and antioxidant activities of rosmary (Rosmarinus officinalis) in alloxan – diabetic rabbits. J Ethnopharmacol  2008; 116:64-73.

12. Peterson M, Simmonds M. Rosmarinic acid. Phytochemistry 2003; 62:121-125.

13. Lee HJ, Cho HS, Seung EP, Lee SY, Kim CS, Kim DK , et al. Rosmarinic acid protects human dopaminergic neuronal cells against hydrogen peroxide – induced apoptosis . Toxicology 2008; 250:109-115.

14.  Erkan N, Ayranci G, Ayranci E. Antioxidant activities of rosmary extract,black seed essential oil,carnosic acid ,rosmarinic acid and sesamol. Food Chem 2008; 110:76-82.

15. Jeanette S, Alex K, Adviye E. Oxidative stress and the use of antioxidants in diabetes. Cardiovasc Diabetol 2005; 4:5-9.

16. Tursun A, Astumi N, Hiroyuki M ,Akio I, Toshitaka N. A natural scavenger of peroxynitrites, protects against impairment of memory induced by Ab25-35. Behav Brain Res 2007; 180:139-145.

17. Liu  B C, Chen Q, Luo DD, Sun J, PhillipsAO, Ruan XZ, Liu NF. Mechanisms of irbesartan in prevention of renal lesion in streptozotocin induced diabetic rats. Acta Pharmacol Sin 2003; 24:67-73.

18. Fernandes NP, Lagishetty CV, Panda VS, Naik SR. An experimental evaluation of the antidiabetic and antilipidemic properties of a standardized Momordica charantia fruit extract. BMC Complement Altern Med 2007; 24:29.

19. Haidara MA, Mikhailidis DP, Rateb MA, Ahmed ZA,Yassin HZ, Ibrahim I.M, et al. Evaluation of the effect of oxidative stress and vitamin E supplementation on renal function in rat with streptozotocin-induced type 1 diabetes. J diabetes and its complications  2009; 23:130-136.

20. Makino T, Ono T, Liu N, Nakamura T, Muso E, Honda G. Suppressive effects of rosmarinic acid on mesangioproliferative glomerulonephritis in rats. Nephron  2002; 92:898-904.

21. Satho K. Serum lipid peroxidation in cerebrovascular disordersdetermined by a new colorimetric method. Clin Chem Acta 1978; 90:37-43.

22. Kim K,Youngki K, Hye P, Hyeon J, Mauer MA .re-evaluation of the renal ablation model of progressive renal disease in rats. J Nephrol 2003; 16:196-202.

23. Gundersen HJ, Bendtsen TF, Korbo L, Marcussen N, Miller A, Nielsen K. Some new, simple and efficient stereological methods and their use in pathological research and diagnosis. APMIS 1988; 96:379-394.

24. Woods L, Ingelfinder J, Nyengaard J, Rasch, R. Maternal protein restriction suppresses the newborn renin- angiotensin system and programs adult hypertension in rats. Pediat Res 2001; 49:460-467.

25. Mayhew TM, Gunderson HJG. ‘If you assume you can make an ass out of u and me': A decade of the disector for stereological counting of particles in 3D space. J Anat 1996; 188:1-15.

26. Woods L, Rasch R. Perinatal ANGII programs adult blood pressure, glomerular number and renal function in rats. Am J Physiol 1998; 275:1593-1599.

27. Nascimento GG, Barbosa FT, Radaeli RF, Cavanal MF, Mello AM, Zaladek GF. Effect of D-alpha tocopherol on tobular nephron acidification by rats with induced diabetes mellitus. Braz J Med Biol Res 2005; 38:1043-1051.

28. Kim SS, Galaher DD, Csallany AS. Vitamin E and probucol reduce urinary lipophilic aldehydes and renal enlargement in streptozotocin –induced diabetic rats. Lipids 2000; 35:1225-1237.

29. Fujita A, Sasaki H, Doi A, Okamoto K, Matsuno S, Furuta H, Nishi M, Nakao T, et al. Ferulic acid prevents pathological and functional abnormalities of the kidney in otsuka long-Evans Tokushima fatty diabetic rats . Diabet Res Clin Pract 2008; 79:11-17.

30. Yoshida M, Kimura H, Kyuki K. Ito M.Effect of combined vitamin E and insulin administration on renal damage in diabetic rats fed a high cholesterol diet. Biol Pharm Bull 2005; 28:2080-2086.

31. Lee EY, Lee MY, Hong SW, Chung CH, Hong SY. Blockade of oxidative stress by vitamin C ameliorates albuminuria and renal sclerosis in experimental diabetic rats. Yonsei Med J 2007; 48:847-855.

32. Winarska K, Malinska D, Szymanski K, Dudziak M, Bryla J. Lipoic acid  ameliorates oxidative stress and renal injury in alloxan diabetic rabbits. Biochemie 2008; 90:450-459 .

33. Kuhad A, Chopra K. Attenuation of diabetic nephropathy by tocotrienol:Involvement of NF KB signaling pathway. Life Sci 2009; 84:296-301.

34. Obrosova II, Fathallah L, Liu E, Nourooz-Zadeh J. Early oxidative stress in the diabetic kidney: effect of DL-α- lipoic acid. Free Rad Biol Med ? 2003; 34:186-195.

35. Vestra M D, Fioretto P. Diabetic nephropathy: renal structural studies in type 1 and type 2 diabetic patients. Internatrional Congress Series 2003; 1253:163–169.

36.  Caramori  ML,  Kim  Y,  Huang  C,  Fish  Aj,  Rich  SS,  Miller  ME,  Russel  G.  Cellular  basis  of  diabetic nephropathy: Study design and renal structure –functional relation ship in patient with long standing type I diabetes. Diabetes 2002; 51:506-513.