SILYMARIN PROTECTS AGAINST COPPER-ASCORBATE INDUCED INJURY TO GOAT CARDIAC MITOCHONDRIA IN VITRO: INVOLVEMENT OF ANTIOXIDANT MECHANISM(S)
Keywords:Silymarin, Antioxidant, Copper-Ascorbate, Goat Heart Mitochondria, Oxidative Stress
Silymarin, 'one of the component of the Milk thistle seeds Silybum marianum (L.) is used in traditional food and medicine in India. In the present study, we investigated the antioxidant activities of Silymarin against copper-ascorbate induced toxic injury to mitochondria obtained from goat heart, in vitro. Incubation of isolated cardiac mitochondria with copper-ascorbate resulted in elevated levels of lipid peroxidation and protein carbonylation of the mitochondrial membrane, a reduced level of mitochondrial GSH and altered status of antioxidant enzymes as well as decreased activities of pyruvate dehydrogenase and the Kreb's cycle enzymes, altered mitochondrial morphology, mitochondrial swelling and di-tyrosine level. All these changes were found to be ameliorated when the cardiac mitochondria were co-incubated with copper-ascorbate and Silymarin, in vitro. Silymarin, in our in vitro experiments, was found to scavenge hydrogen peroxide, superoxide anion free radicals, hydroxyl radicals and DPPH radical, in a chemically defined system, indicating that this compound may provide protection to cardiac mitochondria against copper-ascorbate induced toxic injury through its antioxidant activities. The results of this study suggest that Silymarin may be considered as a future therapeutic antioxidant and may be used singly or as a co-therapeutic in the treatment of diseases associated with mitochondrial oxidative stress.
Davis-Searles PR, Nakanishi Y, Kim NC, Graf TN, Oberlies NH, Wani MC, et al. Milk Thistle and Prostate Cancer:Differential Effects of Pure Flavonolignans from Silybum marianum on Antiproliferative End Points in Human Prostate Carcinoma Cells. J Cancer Res 2005;65(10):4448-57.
EL-Ridy MS, Badawi AA, Safar MM, Mohsen AM. Niosomes as a novel pharmaceutical formulation encapsulating the hepatoprotective drug silymarin. Int J Pharm Pharm Sci 2012;4(1):549-59.
Jashitha M, Chakraborty M, Kamath JV. Pharmacodynamic interaction of quercetin with silymarin against paracetamol induced hepatotoxicity in rats. Int J Pharm Pharm Sci 2013;5(4):104-6.
Flora K, Hahn M, Rosen H, Benner K. Milk Thistle (Silybum marianum) for the therapy of liver diseases. Am J Gastroenterol 1998;93:139-43.
Muriel P, Mourelle M. Prevention by silymarin of membrane alterations in acute CCl4 liver damage. J Appl Toxicol 1990;10:275-9.
Muriel P, Garciapina T, Perez-Alvarez V, Mourelle M. Silymarin protects against paracetamol induced lipid peroxidation and liver damage. J Appl Toxicol 1992;12:439-42.
Fraschini F, Demartini G, Esposti D. Pharmacology of silymarin. J Clin Drug Invest 2002;22:51-65.
Feher J, Deak G, Muzes G, Lang I, Niederland V, Nekam K. Liver protection action of silymarin therapy in chronic alcoholic liver diseases. J Orv Hetil 1989;130:2723-27.
Dhiman RK, Chawla YK. Herbal medicines for liver diseases. J Dig Dis Sci 2005;50:1807-12.
Katiyar SK. Silymarin and skin cancer prevention:anti-inflammatory, antioxidant and immunomodulatory effects (Review). Int J Oncol 2005;26:169-76.
Wagoner J, Negash A, Kane O. Multiple effects of silymarin on the hepatitis C virus lifecycle. J Hepatology 2010;51:1912-21.
Kang JS, Jeon YJ, Kim HM, Han SH, Yang KH. Inhibition of inducible nitric-oxide synthase expression by silymarin in lipopolysaccharide-stimulated macrophages. J Pharmacol Exp Ther 2002;302:138-44.
Kang JS, Park SK, Yang KH, Kim HM. Silymarin inhibits TNF-a-induced expression of adhesion molecules in human umbilical vein endothelial cells. J FEBS Lett 2003;550:89-93.
Alarcon de la Lastra C, Martin MJ, Marhuenda E. Gastric anti-ulcer activity of silymarin, a lipoxygenase inhibitor, in rats. J Pharm Pharmacol 1992;44:929-31.
Babbs CF, Steiner MG. Detection and quantitation of hydroxyl radical using dimethylsulfoxide as a molecular probe. Meth Enzymol 1990;186:137-47.
Bandyopadhyay D, Ghosh G, Bandyopadhyay A, Reiter RJ. Melatonin protects against piroxicam-induced gastric ulceration. J Pineal Res 2004;36:195-203.
Misra HP, Fridovich I. The role of superoxide anion in the autooxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170-5.
Ghosh AK, Mitra E, Dutta M, Mukherjee D, Basu A, Firdaus SB, Bandyopadhyay D, Chattopadhyay A. Protective effect of aqueous bark extract of Terminalia arjuna on Cu2+-ascorbate induced oxidative stress in vitro:involvement of antioxidant mechanism(s). Asian J Pharm Clin Res 2013;6:196-200.
Dutta M, Ghosh AK, Basu A, Bandyopadhyay D, Chattopadhyay A. Protective effect of aqueous bark extract of Terminalia arjuna against copper ascorbate induced oxidative stress in vitro in goat heart mitochondria. Int J Pharm Pharm Sci 2013;5:439-47.
Dutta M, Ghosh AK, Rudra S, Bandyopadhyay D, Guha B, Chattopadhyay A. Human placental mitochondria is a better model for Studies on oxidative stress in vitro:a comparison with Goat heart mitochondria. J Cell Tissue Res 2014;14:3997-4007.
Dutta M, Ghosh AK, Mohan V, Mishra P, Rangari V, Chattopadhyay A, et al. Antioxidant mechanism(s) of protective effects of Fenugreek 4-hydroxyisoleucine and trigonelline enriched fraction (TF4H (28%)) SugahealÂ® against copper-ascorbate induced injury to goat cardiac mitochondria in vitro. J Pharm Res 2014;8(6):798-811.
Fiddler RM. Collaborative study of modified AOAC method of analysis for nitrite in meat and meat products. J AOAC Int 1977;60:594-9.
Griess P. Bemerkungen zu der abhandlung der H.H. Weselsky und Benedikt Ueber einige azoverbindungen.â€ J Chem Ber 1879;12:426-28.
Buege JA, Aust SD. Microsomal lipid peroxidation. J Meth Enzymol 1978;52:302-10.
Sedlak J, Lindsay RH. Estimation of total, protein bound, and non protein sulfhydryl groups in tissue with Ellmanâ€™s reagent. J Anal Biochem 1968;25:192-205.
Levine RL, Williams JA, Stadtman ER, Shacter E. Carbonyl assays for determination of oxidatively modified proteins. J Meth Enzymol 1994;233:346-57.
Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974;47:469-74.
Krohne-Ehrich G, Schirmer RH, Untucht-Grau R. Glutathione reductase from human erythrocytes. Isolation of the enzyme and sequence analysis of the redox active peptide. J Biochem 1977;80:65-71.
Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clinl Med 1967;70:158-69.
Dutta M, Ghosh D, Ghosh AK, Rudra S, Bose G, Dey M, et al. High fat diet aggravates arsenic induced oxidative stress in rat heart and liver. J Food Chem Toxicol 2014;66:262-77.
Chretien D, Pourrier M, Bourgeron T, SÃ©nÃ© M, RÃ¶tig A, Munnich A, et al. An improved spectrophotometric assay of pyruvate dehydrogenase in lactate dehydrogenase contaminated mitochondrial preparations from human skeletal muscles. J Clin Chim Acta 1995;240:129-36.
Duncan MJ, Fraenkel DG. Alpha-ketoglutarate dehydrogenase mutant of Rhizobium meliloti. J Bacteriol 1979;137:415-9.
Veeger C, DerVartanian DV, Zeylemaker WP. Succinate dehydrogenase. J Meth Enzymol 1969;13:81-90.
Giulivi C, Davies KJA. Dityrosine:A marker for oxidatively modified proteins and selective proteolysis. J Meth Enzymol 1994;233:363-71.
Dousset N, Ferretti G, Taus M, Valdiguie P, Curatola G. Fluorescence analysis of lipoprotein peroxidation. J Meth Enzymol 1994;233:459-69.
Halestrap AP, Davidson AM. Inhibition of Ca2(+)-induced large-amplitude swelling of liver and heart mitochondria by cyclo sporin is probably caused by the inhibitor binding to mitochondrial-matrix peptidyl-prolyl cis-trans isomerase and preventing it interacting with the adenine nucleotide translocase. Biochem J 1990;268:153-60.
Lowry OH, Rosenberg NJ, Farr AL, Randall RJ. Protein measurement with Folin-phenol reagent. J Biol Chem 1951;193:265-75.
Sies H. Oxidative Stress:Oxidants and Antioxidants. Academic Press:San Diego California;1991.
Khan SM. Protective effect of black tea extract on the levels of lipid peroxidation and antioxidant enzymes in liver of mice with pesticide-induced liver injury. J Cell Biochem Funct 2006;24:327-32.
Fridovich I. Superoxide dismutases. J Adv Enzymol 1986;58:61-97.
Bhattacharjee R, Sil PC. The protein fraction of Phyllanthus niruri plays a protective role against acetaminophen induced hepatic disorder via its antioxidant properties. J Phytother Res 2006;20:595-601.
Ramakrishnan G, Raghavendran HR, Vinodhkumar R, Devaki T. Suppression of N-nitrosodiethylamine induced hepato carcinogenesis by silymarin in rats. J Chem Biol Interact 2006;161:104-14.
Mayer KE, Myers RP, Lee SS. Silymarin treatment of viral hepatitis:a systematic review. J Viral Hepat 2005;12(6):559-67.
Tanamly MD, Tadros F, Labeeb S, Makld H, Shehata M, Mikhail N, et al. Randomised double-blinded trial evaluating silymarin for chronic hepatitis C in an Egyptian village:study description and 12-month results. J Dig Liver Dis 2004;36(11):752-9.
Polyak SJ, Morishima C, Lohmann V, Pal S, Lee DY, Liu Y, et al. Identification of hepatoprotective flavonolignans from silymarin. J Proc Natl Acad Sci USA 2010;107(13):5995-9.