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Utilization of the Sibininto treat the DMBA Induced Hepatotoxicity and Free-Radical Damage in Norwegian Rat, Rattus norvegicus (L).

Vitthalrao Bhimasha Khyade, Pragati Pramod Shinde, Aarti Sanjay Dhumal


DMBA is abbreviated form of the 7,12-Dimethylbenz[a]anthracene. It is working as  immuno-suppressor. It also serves as a tumor initiator.  It allows for a greatly accelerated rate of growth of tumor and making many cancer studies possible. The plant, Silybum marianum (L) yields pharmacologically active compound named as silbinin. The silbinin deserve documented antioxidant activity. The purpose of this study was to assess this silbinin for protective influence in DMBA induced liver damage. The experimental animals (Norwegian Rat, Rattus norvegicus L) were divided into four groups, like control group (received oral corn oil ); DMBA group (received oral at the rate of 335 mg/kg in the corn oil solution);  Silibinin group (received 100 mg/kg/day silibinin alone orally, every 24 hours for 7 days)  and “Silibinin + DMBA” group (received DMBA plus silibinin). All animals were sacrificed at the end of attempt of schedule of treatment followed by rearing. The tissues used for biochemical assessment were blood and liver. The blood serum was assessed for the parameters that include: Superoxide dismutases (SOD), glutathione peroxidase (GPX), nitric oxide (NO) and myeloperoxidase (MPO) were investigated in serum and liver tissue. In addition, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzyme activities. The homogenate of liver tissue was evaluated histopathologically with Hematoxilin & Eosin dye. The ALT, AST, NO, MPO in serum and NO, MPO in liver tissue were found to be significantly higher in DIMBA treated group in comparison with control group (P < 0.001). In Group: Silibinin + DMBA, serum AST, ALT, NO, MPO levels were significantly lower (P < 0.01), and both serum and tissue SOD activities were significantly higher, compared to DMBA group (P < 0.001). DMBA induced histopathological changes in liver tissue include : severe sinusoidal dilatation, moderate disruption of the radial alignment of hepatocytes around the central vein, severe vacuolization in the hepatocyte cytoplasm, inflammation around central vein and portal region. In rats receiving both DMBA and silibinin, the DMBA induced changes accounted for less sinusoidal dilatation, vacuolization in the hepatocyte cytoplasm and the inflammation around central vein and portal region (P < 0.05). Silibinin exert reduction in the oxidative stress through inducing antioxidant mechanisms, there after showing protective effect against DMBA induced hepatic damage.


Hepatotoxicity; DMBA; Oxidative Stress; Antioxidants; Histopathology; Silbinin

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Klebanoff S J. Myeloperoxidase. ProcAssoc Am Physicians. 1999; 111(5):383–9.

Beyer W, Imlay J, Fridovich I. Superoxide dismutases. Prog Nucleic Acid Res Mol Biol. 1991;40:221–53.

Tutanc M, Arica V, Yilmaz N, Nacar A, Zararsiz I, Basarslan F, et al. Effects of erdosteine on cyclosporin- Miyata M; Furukawa M; Takahashi K; Gonzalez FJ; Yamazoe Y (2001). "Mechanism of 7, 12-Dimethylbenz[a]anthracene-Induced Immunotoxicity: Role of Metabolic Activation at the Target Organ". Jpn J Pharmacol. 86: 302–309. doi:10.1254/jjp.86.302.

Sung YM; He G; Fischer, SM (2005). "Lack of Expression of the EP2 but not EP3 Receptor for Prostaglandin E2 Results in Suppression of Skin Tumor Development". Cancer Res. 65: 9304–9311. doi:10.1158/0008-5472.can-05-1015.

Davis-Searles P, Nakanishi Y, Nam-Cheol K, et al. (2005). "Milk Thistle and Prostate Cancer: Differential Effects of Pure Flavonolignans from Silybummarianum on Antiproliferative End Points in Human Prostate Carcinoma Cells". Cancer Research. 65(10): 4448–57. doi:10.1158/0008-5472.CAN-04-4662.

Saller R, Brignoli R, Melzer J, Meier R (2008). "An updated systematic review with meta-analysis for the clinical evidence of silymarin". ForschendeKomplementärmedizin. 15 (1): 9–20. doi:10.1159/000113648. PMID 18334810. Retrieved 2010-12-14.

Kidd P, Head K (2005). "A review of the bioavailability and clinical efficacy of milk thistle phytosome: a silybin-phosphatidylcholine complex (Siliphos)" (PDF). Alternative Medicine Review. 10 (3): 193–203. PMID 16164374. Retrieved 2010-12-14.

Barzaghi N, Crema F, Gatti G, Pifferi G, Perucca E (1990). "Pharmacokinetic studies on IdB 1016, a silybin- phosphatidylcholine complex, in healthy human subjects". Eur J Drug MetabPharmacokinet. 15: 333–8. PMID 2088770.

Voinovich D, Perissutti B, Grassi M, Passerini N, Bigotto A (2009). "Solid state mechanochemical activation of Silybummarianum dry extract with betacyclodextrins: Characterization and bioavailability of the coground systems". Journal of Pharmaceutical Sciences. 98 (11): 4119–29. doi:10.1002/jps.21704. PMID 19226635.

Kosina P, Kren V, Gebhardt R, Grambal F, Ulrichová J, Walterová D (2002). "Antioxidant properties of silybin glycosides". PhytotherapyResearch : PTR. 16 Suppl 1: S33–9. doi:10.1002/ptr.796. PMID 11933137.

Zhou S, Lim LY, Chowbay B (2004). "Herbal modulation of P-glycoprotein". Drug Metabolism Reviews. 36 (1): 57–104. doi:10.1081/DMR-120028427. PMID 15072439.

Wu JW, Lin LC, Tsai TH (2009). "Drug-drug interactions of silymarin on the perspective of pharmacokinetics". Journal of Ethnopharmacology. 121 (2): 185–93. doi:10.1016/j.jep.2008.10.036. PMID 19041708. Retrieved 2010-12-14.

Thomas W. Flaig; Daniel L. Gustafson; Lih-Jen Su; Joseph A. Zirrolli; Frances Crighton; Gail S. Harrison; A. Scott Pierson; Rajesh Agarwal; L. Michael Glodé (2007). "A phase I and pharmacokinetic study of silybin-phytosome in prostate cancer patients". Investigational New Drugs. 25 (2): 139–146. doi:10.1007/s10637-006-9019-2.

Fraschini F, Demartini G, Esposti D (2002). "Pharmacology of Silymarin". Clinical Drug Investigation. 22 (1): 51–65. doi:10.2165/00044011-200222010-00007.

Hahn G, Lehmann HD, Kürten M, Uebel H, Vogel G (1968). "On the pharmacology and toxicology of silymarin, an antihepatotoxic active principle from Silybummarianum (L.) gaertn". Arzneimittelforschung. 18 (6): 698–704. PMID 5755807.

Bosch-Barrera J, Menendez JA (2015). "Silibinin and STAT3: A natural way of targeting transcription factors for cancer therapy". Cancer Treat. Rev. (Review). 41 (6): 540–6. doi:10.1016/j.ctrv.2015.04.008. PMID 25944486.

Singh, Rana P.; Agarwal, Rajesh (September 2009). "Cosmeceuticals and silibinin". Clinics in Dermatology. 27 (5): 479–484. doi:10.1016/j.clindermatol.2009.05.012. PMC 2767273. PMID 19695480.

Trappoliere M, Caligiuri A, Schmid M, Bertolani C, Failli P, Vizzutti F, et al. Silybin, a component of sylimarin, exerts anti-inflamma¬tory and anti-fibrogenic effects on human hepatic stellate cells. J Hepatol. 2009;50(6):1102–11.

Trouillas P, Marsal P, Svobodova A, Vostalova J, Gazak R, Hrbac J, et al. Mechanism of the antioxidant action of silybin and 2,3-dehy¬drosilybin flavonolignans: a joint experimental and theoretical study. J PhysChem A. 2008;112(5):1054–63.

Gazak R, Walterova D, Kren V. Silybin and silymarin--new and emerging applications in medicine. Curr Med Chem. 2007; 14(3):315–38.

Prabu SM, Muthumani M. Silibinin ameliorates arsenic induced nephrotoxicity by abrogation of oxidative stress, inflammation and apoptosis in rats. MolBiol Rep. 2012;39(12):11201–16.

Andreoli TE, Carpenter CCJ, Cecil RLF. Cecil Essentials of Medicine. 3rd edMichigan: Saunders; 1995.

A-induced nephrotoxicity. Hum ExpToxicol. 2012;31(6):565–73.

Peresleni T, Noiri E, Bahou WF, Goligorsky MS. Antisense oligode¬oxynucleotides to inducible NO synthase rescue epithelial cells from oxidative stress injury. Am J Physiol. 1996;270(6 Pt 2):F971–7.

Arthur JR. The glutathione peroxidases. Cell Mol Life Sci. 2000;57(13-14):1825–35.

Shah MD, Iqbal M. Diazinon-induced oxidative stress and renal dysfunction in rats. Food ChemToxicol. 2010;48(12):3345–53.

Yilmaz N, Yilmaz M, Altuntas I. Diazinon-induced brain tox¬icity and protection by vitamins E plus C. ToxicolInd Health. 2012;28(1):51–7.

Teimouri F, Amirkabirian N, Esmaily H, Mohammadirad A, Ali¬ahmadi A, Abdollahi M. Alteration of hepatic cells glucose me¬tabolism as a non-cholinergic detoxication mechanism in coun¬teracting diazinon-induced oxidative stress. Hum ExpToxicol. 2006;25(12):697–703.

Nakagawa Y, Moore G. Role of mitochondrial membrane perme¬ability transition in p-hydroxybenzoate ester-induced cytotoxic¬ity in rat hepatocytes. BiochemPharmacol. 1999;58(5):811–6.

Yehia MA, El-Banna SG, Okab AB. Diazinon toxicity affects histo¬physiological and biochemical parameters in rabbits. ExpToxicolPathol. 2007;59(3-4):215–25.

Alp H, Aytekin I, Esen H, Basarali K, Kul S. Effects of caffeic acid phenethyl ester, ellagic acid, sulforaphane and curcumin on diazi¬non induced damage to the lungs, liver and kidneys in an acute toxicity rat model. KafkasUniv Vet FakDerg. 2011; 17(6):927–33.

Giray B, Gurbay A, Hincal F. Cypermethrin-induced oxidative stress in rat brain and liver is prevented by vitamin E or allopuri¬nol. ToxicolLett. 2011;118(3):139–46.

Akturk O, Demirin H, Sutcu R, Yilmaz N, Koylu H, Altuntas I. The effects of diazinon on lipid peroxidation and antioxidant en¬zymes in rat heart and ameliorating role of vitamin E and vita¬min C. Cell BiolToxicol. 2007;22(6):455–61.

Mehta A, Verma RS, Srivastava N. Chlorpyrifos induced altera¬tions in the levels of hydrogen peroxide, nitrate and nitrite in rat brain and liver. Pesticide Biochem Physiol. 2009;94(2-3):55–9.

Klebanoff SJ. Myeloperoxidase: friend and foe. J Leukoc Biol. 2005;77(5):598–625.

Heinecke JW, Li W, Francis GA, Goldstein JA. Tyrosyl radical gener¬ated by myeloperoxidase catalyzes the oxidative cross-linking of proteins. J Clin Invest. 1993;91(6):2866–72.

Messarah M, Amamra W, Boumendjel A, Barkat L, Bouasla I, Ab¬dennour C, et al. Ameliorating effects of curcumin and vitamin E on diazinon-induced oxidative damage in rat liver and erythro¬cytes. ToxicolInd Health. 2013;29(1):77–88.

Sozmen M, Devrim AK, Tunca R, Bayezit M, Dag S, Essiz D. Protec¬tive effects of silymarin on fumonisin B(1)-induced hepatotoxic¬ity in mice. J Vet Sci. 2014;15(1):51–60.

Lee AG, Malcolm East J, Balgavy P. Interactions of insecticides with biological membranes. Pesticide Sci. 1991;32(3):317–27.

Vitthalrao B. Khyade and Rhidim Jiwan P. Sarwade (2014). Influence of Moracin on DMBA – TPA induced cancer in the skin of mice, Musmusculus (L). Recent Trends in Zoology (Pages: 113-133). Editor: Dr. R. K. Kasar ; Publisher: Dr. L. S. Matkar (Principal, New Arts, Commerce and Science College, Shevgaon Dist. Ahmednagar – 414502 (M.S.) India. ISBN: 978-93-84916-68-8.

Erlejman AG, Verstraeten SV, Fraga CG, Oteiza PI. The interaction of flavonoids with membranes: potential determinant of flavo¬noid antioxidant effects. Free Radic Res. 2004;38(12):1311–20.

Cacciapuoti F, Scognamiglio A, Palumbo R, Forte R, Cacciapuoti F. Silymarin in non alcoholic fatty liver disease. World J Hepatol. 2013;5(3):109–13.

Lazarević-Pašti T, Čolović M, Savić J, Momić T, Vasić V. Oxidation of diazinon and malathion by myeloperoxidase. Pesticide Biochem Physiol. 2011;100(2):140–4.

van Wenum E, Jurczakowski R, Litwinienko G. Media effects on the mechanism of antioxidant action of silybin and 2,3-dehydro¬silybin: role of the enol group. J Org Chem. 2013;78(18):9102–12.

Sirmali M, Solak O, Tezel C, Sirmali R, Ginis Z, Atik D, et al. Com¬parative analysis of the protective effects of caffeic acid pheneth¬yl ester (CAPE) on pulmonary contusion lung oxidative stress and serum copper and zinc levels in experimental rat model. Biol Trace Elem Res. 2013;151(1):50–8.

Vitthalrao B. Khyade1 and Aziz Sancer (2016).Treating the 7,12-dimethylbenz(a)anthracene (DMBA) induced buccal pouch carcinoma in Syrian hamster, Mesocricetusauratus(L) with ethanolic extractives of leaves of mulberry, Morus alba (L).World Scientific news 30 (2016): 1-13.

Ezhilarasan D, Karthikeyan S, Vivekanandan P. Ameliorative effect of silibinin against N-nitrosodimethylamine-induced hepatic fi¬brosis in rats. Environ ToxicolPharmacol. 2012; 34(3):1004–13.

Vitthalrao B. Khyade (2016).Impact of Moracin on Hydrogen Peroxide induced oxidative stress in feline fibroblasts.International Academic Journal of Innovative Research. Vol. 3No.10 : 45 - 59 . ISSN 2454-390X.


Vitthalrao B. Khyade and Ujwala D. Lonkar( 2013 ):Effect of Moracin on DMBA – TPA induced cancer in mice, Musmusculus(L). 2013.Annals of Plant Science Vol. 2 No. 10 (2013):412 – 419.

Shadnia S, Ashrafivand S, Mostafalou S, Abdollahi M. N-acetyl¬cysteine a Novel Treatment for Acute Human Organophosphate Poisoning. Int J Pharmacology. 2011;7(6).


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