Dimethylnitrosamine (DMN)-induced fibrotic rats: effect of Vernonia amygdalina on extracellular matrix and Hepatic/lysosomal integrity

  • Abstract
  • Keywords
  • References
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  • Abstract

    Background: Hepatic fibrosis is the accumulation of extracellular matrix protein, or scar, in response to acute or chronic liver injury. This study investigated the effect of ethanolic leaf extract of Vernonia amygdalina on extracellular matrix protein: hyaluronic acid (HA), liver synthetic molecules: total protein (TP), albumin (ALB), total bilirubin (TB) as well as lysosomal membrane stability: acid phosphatase (ACP) in dimethylnitrosamine (DMN)-induced fibrotic rats.

    Methods: Wistar albino male rats were intraperitoneally injected with 10 mg/kg DMN on first three days a week for two weeks. Ethanolic leaf extracts of Vernonia amygdalina (200 mg /kg) was administered simultaneously by oral gavage daily for two weeks. All rats were sacrificed after 24 hours of last administration by cardiac puncture, and blood collected from the ocular vein. Analysis of serum ACP and LDH activities with those of the concentrations of HA, ALB, TP and TB were carried out.

    Results: Administration of DMN to rats significantly increased HA and TB concentration and the activities of ACP and LDH (p<0.05) in the serum while it significantly reduced (p<0.05) serum TP and ALB concentrations when compared with controls. However, simultaneous administration of ethanolic leaf extracts of Vernonia amygdalina with DMN significantly (p<0.05) reversed these changes.

    Conclusion: This study shows that Vernonia amygdalina possesses hepatoprotective, lysosomal membrane stabilizing and anti-fibrotic properties may be due to its antioxidant and phytochemical constituents.

  • Keywords

    Dimethylnitrosamine; Extracellular Matrix; Fibrosis, Liver; Vernonia Amygdalina.

  • References

      [1] Ahmed OM (2001): Histopathological and biochemical evaluation of liver and kidney lesions in streptozotocin diabetic rats treated with glimepiride and various plant extracts. Journal Union Arab Bioliogy 16A, 585-625.

      [2] Akah PA & Okafor CL (2006). Blood sugar lowering effect of Vernonia amygdalina Del, in an experimental rabbit model. Phytotherapy Research 6, 171-173. http://dx.doi.org/10.1002/ptr.2650060318.

      [3] Akanji MA, Nafiu M.O, & Yakubu MT. (2008). Enzyme Activities and Histopathology of Selected Tissues in Rats Treated with Potassium bromate. African Journal of Biomedical Research 11, 87 – 95.

      [4] Ala–Kokko L, Pihlajaniemi T., Myers JC, & Kivirikko KI, & Savolainen ER. (1987). Gene expression of type I, III and IV collagens in hepatic fibrosis induced by dimethylnitrosamine in the rat. Biochemistry Journal 244, 75–79. http://dx.doi.org/10.1042/bj2440075.

      [5] Babalola OO, Anetor JI, & Adeniyi FA. (2001). Amelioration of carbon tetrachloride induced hepatotoxicity of terpenoid extract from leaves of Vernonia amygdalina. African Journal of Medical Sciences 30(1-2), 91-93.

      [6] Chichibu K, Matsuura, T, & Shichijo S. (1989). Assay of serum hyaluronic acid in clinical application. Clinica Chimica Acta 181, 317-324. http://dx.doi.org/10.1016/0009-8981(89)90237-4.

      [7] Collins, AJ. & Lewis DA. (1971). Lysosomal enzyme level in blood of arthritic rats. Biochemistry and Pharmacology 28, 251-253. http://dx.doi.org/10.1016/0006-2952(71)90496-5.

      [8] Dasgupta T, Banerjee S, Yadava PK & Rao AR. (2004). Chemopreventive potential of Azadirachta indica (Neem) leaf extract in murine carcinogenesis model system. Journal Ethnopharmacology 92, 23. http://dx.doi.org/10.1016/j.jep.2003.12.004.

      [9] EL-Maragy SA, Rizk SA. & EL-Sawalhi MM. (2009): Hepatoprotective potential of crocin and curcumine against iron overload- iduced biochemical alterations in rat. African Journal of Biochemistry Research 3(5), 215-221.

      [10] El-Zayat EM. (2007). Isoenzyme pattern and activity in oxidative stress-induced hepatocarcinogenesis: the protective role of selenium and Vitamin E. Research Journal Medicine & Medical Sciences 2, 62–71.

      [11] Farber E. (1996). Orientation in Liver Toxicity by Drugs, Handbook of Experimental Pharmacology, Springer-Verlag, Berlin, Germany. http://dx.doi.org/10.1007/978-3-642-61013-4_1.

      [12] Fasano M, Curry S, Terreno E, Galliano M, Fanali G, & Narciso P, (2005). The extraordinary ligand binding properties of human serum albumin. IUBMB Life. 57, 787–796. http://dx.doi.org/10.1080/15216540500404093.

      [13] Fraser JR, Alcorn D, Laurent TC, Robinson AD, & Ryan GB. (1985). Uptake of circulating hyaluronic acid by the rat liver. Cellular localization in situ. Cell Tissue Research 242, 505–510. http://dx.doi.org/10.1007/BF00225415.

      [14] Fraser JRE, Laurent TC, & Laurent UBG (1997). Hyaluronan: its nature, distribution, functions and turnover. Journal of Internal Medicine 242(1), 27-33. http://dx.doi.org/10.1046/j.1365-2796.1997.00170.x.

      [15] Ganger S.C., Sandhir R., Rai D.V., and Koul A., (2006). Preventive effects of Azadirachta indica on benzo(a)pyrene DNA adduct formation in murine forestomach and hepatic tissues. Phytotherapy Research 20, 88-89.

      [16] George J, Suguna L, Jayalakshmi R., & Chandrakasan G. (2006) Efficacy of Silymarin and Curcumin on Diethylnitrosamine induced liver fibrosis in rats. Biomedicine 26(3-4), 18-26.

      [17] George J, (2008). Elevated serum β-glucuronidase reflects hepatic lysosomal fragility following toxic liver injury in rats. Biochemistry and Cell Biology 86, 235–243. http://dx.doi.org/10.1139/O08-038.

      [18] George J & Chandrakasan G (1997). Lactate dehydrogenase isoenzymes in dimethylnitrosamine induced hepatic fibrosis in rats. Journal of Clinical Biochemistry and Nutrition 22: 51–62. http://dx.doi.org/10.3164/jcbn.22.51.

      [19] George J, Rao KR, Stern R, & Chandrakasan G (2001). Dimethylnitrosamine-induced liver injury in rats: the early deposition of collagen. Toxicology 156(2-3), 129-138. http://dx.doi.org/10.1016/S0300-483X(00)00352-8.

      [20] Greige-Gerges H, Khalil RA, Chahine R, Haddad C & Harb W (2007). Effect of cucurbitacins on bilirubin-albumin binding in human plasma. Life Sciences 80, 579-585. http://dx.doi.org/10.1016/j.lfs.2006.10.005.

      [21] Hartley JL, Brown RM, & Tybulewicz A (2006). Hyaluronic acid predicts hepatic fibrosis in children with hepatic disease. Journal of Pediatric Gastroenterology and Nutrition 43, 217-21. http://dx.doi.org/10.1097/01.mpg.0000228121.44606.9f.

      [22] Hasegawa T, Sasaki T, Kimura T, Hoki M, Okada A, Mushiake S, Yagi M, & Imura K (2000). Measurement of serum hyaluronic acid as a sensitive marker of liver fibrosis in biliary atresia. Journal of Pediatric Surgery 35, 1643–1646. http://dx.doi.org/10.1053/jpsu.2000.18342.

      [23] Huang W, Zhang J Chua SS Qatanani M & Han Y (2003). Induction of bilirubin clearance by the Constitutive Androstane Receptor (CAR). PNAS 100, 4156-4161. http://dx.doi.org/10.1073/pnas.0630614100.

      [24] Izevbige EB, Bryant TL, & Walker A (2004). A novel natural hibitor of extracellular signal regulated kinases and human breast cancer cell growth. Experimental Biology and Medicine 229(2), 163-169.

      [25] Kambizi L. & Afolayan AJ (2001). An ethnobotanical study of plants used for the treatment of sexually transmitted disease (njovher) in Guruve District, Zimbabwe. Journal of Ethnopharmacology 77, 5-9. http://dx.doi.org/10.1016/S0378-8741(01)00251-3.

      [26] Kobyashi H, Horikoshi K, and Yamataka A, (1999). Hyaluronic acid: a specific prognostic indicator of hepatic damage in biliary atresia. Journal Pediatric Surgery 34, 1791-94. http://dx.doi.org/10.1016/S0022-3468(99)90314-7.

      [27] Liu SQ, Yu JP, Chen HL, Luo HS, Chen SM & Yu HG (2006). “Therapeutic effects and molecular mechanisms of Ginkgo biloba Extract on liver fibrosis in rats,” The American Journal of Chinese Medicine 34 (1), 99–114. http://dx.doi.org/10.1142/S0192415X06003679.

      [28] Mio K, Carrette O, Maibach HI & Stern R (2000). Evidence that the serum inhibitor of hyaluronidase may be a member of the inter-alpha-inhibitor family. Journal of Biology and Chemistry 275, 32413–32421. http://dx.doi.org/10.1074/jbc.M005428200.

      [29] Montazeri G, Estakhri A, & Mohamadnejad M (2005). Serum hyaluronate as a non-invasive marker of hepatic fibrosis and inflammation in HBeAg-negative chronic hepatitis B. BMC Gastroenterology 5, 32. http://dx.doi.org/10.1186/1471-230X-5-32.

      [30] Muthulingam M, Mohandoss P, Indra N, & Sethupathy S (2010). Antihepatotoxic efficacy of Indigofera tinctoria on paracetamol-induced liver damage in rats. International Journal of Pharmacy and Biomedical Research 1(1), 13-18.

      [31] Nadia N, Hassan, Firyal HA, & Hala SA (2013). The Values of Hyaluronic Acid and as a Marker of Cirrhosis in Children with Chronic Liver Diseases. Iraqi Journal of Medical Sciences 12(1), 70-77.

      [32] Ostrow JD., Pascolo L, Shapiro SM & Tiribelli, C (2003). New concepts in bilirubin encephalopathy. European Journal of Clinical Investment 33, 988-997. http://dx.doi.org/10.1046/j.1365-2362.2003.01261.x.

      [33] Patel K, Lajoie A, Heaton S, Pianko S, Behling CA, Bylund D, Pockros PJ, Blatt LM, Conrad A, & McHutchison JG (2003). Clinical use of hyaluronic acid as a predictor of fibrosis change in hepatitis C. Journal of Gastroenterology and Hepatology 18, 253–257. http://dx.doi.org/10.1046/j.1440-1746.2003.02930.x.

      [34] Pillay CS, Elliott E, & Dennison C (2002). Endolysosomal proteolysis and its regulation. Biochemistry Journal 363(3), 417-429. http://dx.doi.org/10.1042/bj3630417.

      [35] Regassa A (2000). The use of herbal preparations for tick control in western Ethiopia. Journal South-African Veterinary Association 71, 240-243. http://dx.doi.org/10.4102/jsava.v71i4.722.

      [36] [36] Saegusa S, Isaji S, & Kawarada Y (2002). Changes in serum hyaluronic acid levels and expression of CD44 and CD44 mRNA in hepatic sinusoidal endothelial cells after major hepatectomy in cirrhotic rats. World Journal of Surgery 26, 694–699. http://dx.doi.org/10.1007/s00268-001-0292-0.

      [37] Sharma V & Singh M (2014). Attenuation of N-nitrosodimethylamine induced hepatotoxicity by Operculina turpethum in Swiss Albino mice. Iran Journal of Basic Medical Sciences 17, 73-80.

      [38] Shin M & Moon J (2010). Effect of dietary supplementation of grape skin and seeds on liver fibrosis induced by dimethylnitrosamine in rats. Nutrition Research and Practice 4(5), 369-374. http://dx.doi.org/10.4162/nrp.2010.4.5.369.

      [39] Ueno T, Inuzuka S, Torimura T, Tamaki S, Koh H, Kin M, Minetoma T, Kimura Y, Ohira H, Sata M, Yoshida H, & Tanikawa K (1993). Serum hyaluronate reflects hepatic sinusoidal capillarization. Gastroenterology 105, 475–481.

      [40] Usunobun U, Okolie PN & Eze GI (2015a). Effect of Vernonia amygdalina on some biochemical indices in Dimethylnitrosamine (DMN)-induced liver injury in rats. International Journal of Animal Biology 1(4), 99-105.

      [41] Usunobun U, Okolie NP & Eze IG (2015b). Inhibitory Effect of Vernonia amygdalina on Dimethylnitrosamine (DMN)-induced Liver Fibrosis in Rats. International Journal of clinical Pharmacology and Toxicology 4(4), 179-184.

      [42] Usunobun U (2014). Antihepatotoxic efficacy of Vernonia amygdalina ethanolic leaf extract on Dimethylnitrosamine (DMN)-induced liver damage in rats. International Journal of Healthcare and Biomedical Research. 03(01), 89-98

      [43] Usunobun U & Okolie NP (2015). Phytochemical, trace and mineral composition of Vernonia amygdalina leaves. International Journal of Biological and Pharmaceutical Research 6(5), 393-399.

      [44] Vogel AI (1971). A textbook of practical organic Chemistry including qualitative organic analysis. Longman group limited, London. Pp 426.

      [45] Vrochides D, Papanikolaou V, Pertoft H, Antoniades AA & Heldin, P (1996). Biosynthesis and degradation of hyaluronan by nonparenchymal liver cells during liver regeneration. Hepatology 23, 1650–1655. http://dx.doi.org/10.1002/hep.510230648.




Article ID: 5785
DOI: 10.14419/ijpt.v4i1.5785

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