The Protective Effect of Garlic Extract againstAcetaminophen-Induced Loss of Mitochondrial MembranePotential in Freshly Isolated Rat Hepatocytes Effect of garlic extract against acetaminophen cytotoxicity
Iranian Journal of Pharmaceutical Sciences,
Vol. 5 No. 3 (2009),
1 July 2009
,
Page 141-150
https://doi.org/10.22037/ijps.v5.41209
Abstract
Overdose of acetaminophen causes severe hepatic necrosis in humans and experimental animals. Studies on its hepatotoxicity remain a very active area since some of current data are still uncertain. In this study, freshly isolated rat hepatocytes were used to determine the effects of garlic extract and its component, allicin on the acetaminophen-induced cell cytotoxicity and to compare with the effect of N-acetyl cysteine as a standard treatment. Garlic extract was prepared via a standardmethod and its allicin and allyl mercaptan contents were determined using analytical and preparative high performance liquid chromatography (HPLC). Rat hepatocytes were isolated using collagenase perfusion and mitochondrial membrane potential and cell cytotoxicity were determined using Rhodamine 123 fluorescence and trypan blue exclusion, respectively. Inclusion of garlic extract and/or N-acetyl cysteine resulted in a reduction in the loss of mitochondrial membrane potential as well as cell death which occurred after acetaminophen addition and therefore,illustrated considerable hepatoprotective effects without significant differences between two treatments. In contrast, pure allicin was not effective significantly. The hepatoprotective effects of garlic extract may be due to the compounds other than allicin such as allyl mercaptan, as allicin has been shown to transform to allyl mercaptan as a major metabolite.
Keywords:
- Allicin
- Cytotoxicity
- Garlic extract
- Hepatocytres
- Mitochondrialmembrane potential
How to Cite
Mahdieh Anoush, Mohammad Ali Eghbal, Hossein Hamzeiy, Fatemeh Fathiazad, & Nazli Saeedi Kouzehkonani. (2009). The Protective Effect of Garlic Extract againstAcetaminophen-Induced Loss of Mitochondrial MembranePotential in Freshly Isolated Rat Hepatocytes: Effect of garlic extract against acetaminophen cytotoxicity. Iranian Journal of Pharmaceutical Sciences, 5(3), 141–150. https://doi.org/10.22037/ijps.v5.41209
References
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[25]Lemar KM, Passa O, Aon MA, Cortassa S, Müller CT, Plummer S, O'Rourke B, Lloyd D. Allyl alcohol and garlic (Allium sativum) extract produce oxidative stress inCandida albicans.Microbiology 2005; 151: 3257-65.
[26]Chu Q, Ling MT, Feng H, Cheung HW, Tsao SW, Wang X, Wong YC. Anovel anticancer effect of garlic derivatives: Inhibition of cancer cell invasion through restoration of E-cadherinexpression. Carcinogenesis2006; 27: 2180-9.
[27]Seki T, Ozono T, Hosono-Fukao T, Inada K,Tanaka R, Ogihara J, Ariga T. Anticancer effects of diallyl trisulfide derived from garlic. Asia PacJ Clin Nutr2008; 17 Suppl 1: 249-52.
[28]Gebhardt R. Multiple inhibitory effects of garlic extracts on cholesterol biosynthesis in hepatocytes.Lipids1993; 28: 613-9.
[29]Rousar T, Koucera O, Krivakova P, Lotkova H,Kandar R, Muzakova V, Cervinkova Z. Evaluation of oxidative status in acetaminophen treated rat hepatocytes in culture.Physiol Res(in press).
[30]Sumioka I, Mansura T, Yamada K.Acetaminophen induced hepatotoxicity: Still an important issue. Yonago Acta Medica2004; 47:17-28.
[31]Wargovich MJ. Diallylsulfide and allylmethylsul-fide are uniquely effective among organosulfur compounds in inhibiting CYP2E1 protein in animal models. J Nutr2006; 136: 832-4S.
[32]Amagase H, Petesch BL, Matsuura H, Kasuga S,Itakura Y. Intake of garlic and its bioactive components. J Nutr2001; 131: 955-62S.
[33]Lawson, LD, Ransom DK, Huges BG. Inhibition of whole blood platelet aggregation by compounds in garlic clove extracts and commercial garlic products. Thromb Res1992; 65: 141-56.
[34]Song K, Milner JA. The influence of heating on the anticancer properties of garlic. J Nutr2001;131: 1054-7S.
[35]Zeng, T, Zhang CL, Zhu ZP, Yu LH, Zhao XL, Xie KQ. Diallyl trisulfide (DATS) effectively attenuated oxidative stress-mediated liver injury and hepatic mitochondrial dysfunction in acute ethanol-exposed mice.J Toxicol2008; 252: 86-91.
[36]Cong Z, Duquet S, Yao-Xia Z. Effects of combined use of diallyl disulfide and N-acetyl-cysteine on acetaminophen hepatotoxicity in ß-naphthoflavone pretreated mice. WJG1998;4: 153-7.
[2]El-Shenawy SM, Hassan NS. Comparative evaluation of the protective effect of selenium and garlic against liver and kidney damage induced by mercury chloride in the rats. Pharmacol Rep 2008; 60: 199-208.[3]Lawson LD. Bioactive organosulfur compounds of garlic and garlic products- role in reducing blood-lipids. Acs Symposium Series1993; 534:306-30.
[4]Koch H. The long path toward ‘odorless garlic’.Pharm Unserer Zeit1996; 25: 186-91.
[5]Egen-Schwind C, Eckard R, Kemper FH.Metabolism of garlic constituents in the isolated perfused rat liver. Planta Med1992; 58: 301-5.
[6]Xu S, Simon Cho BH. Allyl mercaptan, a major metabolite of garlic compounds, reduces cellular cholesterol synthesis and its secretion in Hep-G2 cells. J Nutr Biochem1999; 10: 654-9.
[7]Aleksunes LM, Campion SN, Goedken MJ,Manautou JE. Acquired resistance to acetaminophen hepatotoxicity is associated with induction of multidrug resistance-associated protein 4 (Mrp4) in proliferating hepatocytes.Toxicol Sci2008; 104: 261-73.
[8]Amar J, Schiff ER. Acetaminophen safety and hepatotoxicity-where do we go from here? Expert Opin Drug Saf 2007; 6: 341-55.
[9]Dai G, He L, Chu N, Wan YJ. Acetaminophen metabolism does not contribute to gender difference in its hepatotoxicity in mouse. ToxicolSci2006; 92: 33-41.
[10]Corcoran GB, Wong BK. Role of glutathione in prevention of acetaminophen-induced hepatotoxicity by N-acetyl-L-cysteine in vivo:Studies with N-acetyl-D-cysteine in mice. JPharmacol Exp Ther1986; 238: 54-61.
[11]James L, Mayeux R, Hinson JA. Acetaminophen-induced hepatotoxicity. Drug Metab Dispos2003;31: 1499-506.
[12]Ruepp SU, Tongue RP, Shaw J, Wallis N, Pognan F. Genomics and proteomics analysis of acetaminophen toxicity in mouse liver. Toxicol Sci 2002; 65: 135-50.
[13]Mitchell JR. Host susceptibility and acetaminophen liver injury.Ann Intern Med 1977;87: 377-8.
[14]Donnelly PJ, Walker RM, Racz WJ. Inhibition of mitochondrial respirationin vivois an early event in acetaminophen-induced hepatotoxicity.Arch Toxicol 1994; 68: 110-8.
[15]Baluchnejadmojarad T, Roghani M, Homayounfar H, Hosseini M. Beneficial effect of aqueous garlic extract on the vascular reactivity of streptozotocin-diabetic rats. J Ethnopharmacol2003; 85: 139-44.[16]Vargas I, Corrales C, Martinez MA. Compounds derived from garlic as bud induction agents in organic farming of table grape. Chilean J Agri Res2008; 68: 94-9.
[17]Eghbal MA, Pennefather PS, O'Brien PJ. H2S cytotoxicity mechanism involves reactive oxygen species formation and mitochondrial depolarisation. Toxicology2004; 203: 69-76.
[18]Moldeus P, Mogberg J, Orrenius S. Isolation and use of liver cells. Methods Enzymol 1978; 52: 60-71.
[19]Mingatto FE, Rodrigues T, Pigoso AA, Uyemura SA, Curti C, Santos AC. The critical role of mitochondrial energetic impairment in the toxicity of nimesulide to hepatocytes. J Pharmacol ExpTher2002; 303: 601-7.[20]Eghbal MA, Tafazoli S, Pennefather PS, O'Brien PJ. Peroxidase catalysed formation of cytotoxic prooxidant phenothiazine free radicals at physiological pH. Chem Biol Interact2004; 151:43-51.
[21]Marvasi L, Zaghini A, Gervasi PJ, Vaccaro E,Chirulli V. Induction of cytochrome P450 1A by beta-naphto-flavon within the pig liver and central nervous system. Vet Res Commun2006; 30: 333-6.
[22]Sun X, Ku DD. Allicin in garlic protects against coronary endothelial dysfunction and right heart hypertrophy in pulmonary hypertensive rats. Am J Physiol Heart Circ Physiol2006; 291: 2431-8.
[23]O'Gara EA, Hill DJ, Maslin DJ. Activities of garlic oil, garlic powder, and their diallyl constituents against Helicobacter pylori. Appl Environ Microbiol2000; 66: 2269-73.
[24]Ross ZM, O'Gara EA, Hill DJ, Sleightholme HV,Maslin DJ. Antimicrobial properties of garlic oil against human enteric bacteria: Evaluation of methodologies and comparisons with garlic oil sulfides and garlic powder. Appl Environ Microbiol2001; 67: 475-80.
[25]Lemar KM, Passa O, Aon MA, Cortassa S, Müller CT, Plummer S, O'Rourke B, Lloyd D. Allyl alcohol and garlic (Allium sativum) extract produce oxidative stress inCandida albicans.Microbiology 2005; 151: 3257-65.
[26]Chu Q, Ling MT, Feng H, Cheung HW, Tsao SW, Wang X, Wong YC. Anovel anticancer effect of garlic derivatives: Inhibition of cancer cell invasion through restoration of E-cadherinexpression. Carcinogenesis2006; 27: 2180-9.
[27]Seki T, Ozono T, Hosono-Fukao T, Inada K,Tanaka R, Ogihara J, Ariga T. Anticancer effects of diallyl trisulfide derived from garlic. Asia PacJ Clin Nutr2008; 17 Suppl 1: 249-52.
[28]Gebhardt R. Multiple inhibitory effects of garlic extracts on cholesterol biosynthesis in hepatocytes.Lipids1993; 28: 613-9.
[29]Rousar T, Koucera O, Krivakova P, Lotkova H,Kandar R, Muzakova V, Cervinkova Z. Evaluation of oxidative status in acetaminophen treated rat hepatocytes in culture.Physiol Res(in press).
[30]Sumioka I, Mansura T, Yamada K.Acetaminophen induced hepatotoxicity: Still an important issue. Yonago Acta Medica2004; 47:17-28.
[31]Wargovich MJ. Diallylsulfide and allylmethylsul-fide are uniquely effective among organosulfur compounds in inhibiting CYP2E1 protein in animal models. J Nutr2006; 136: 832-4S.
[32]Amagase H, Petesch BL, Matsuura H, Kasuga S,Itakura Y. Intake of garlic and its bioactive components. J Nutr2001; 131: 955-62S.
[33]Lawson, LD, Ransom DK, Huges BG. Inhibition of whole blood platelet aggregation by compounds in garlic clove extracts and commercial garlic products. Thromb Res1992; 65: 141-56.
[34]Song K, Milner JA. The influence of heating on the anticancer properties of garlic. J Nutr2001;131: 1054-7S.
[35]Zeng, T, Zhang CL, Zhu ZP, Yu LH, Zhao XL, Xie KQ. Diallyl trisulfide (DATS) effectively attenuated oxidative stress-mediated liver injury and hepatic mitochondrial dysfunction in acute ethanol-exposed mice.J Toxicol2008; 252: 86-91.
[36]Cong Z, Duquet S, Yao-Xia Z. Effects of combined use of diallyl disulfide and N-acetyl-cysteine on acetaminophen hepatotoxicity in ß-naphthoflavone pretreated mice. WJG1998;4: 153-7.
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