مطالعه خواص آنتی اکسیدان عصاره گیاه سنبل بیابانی در هپاتوسیتهای رت

Abstract

Eremostachys laciniata (L) Bunge (family Lamiaceae) is one of the 15 endemic Iranian herbs of the genus Eremostachys. Eremostachys laciniata is a rich source of flavonoids. Some species of E. laciniata are used in traditional medicine for the treatment of allergies, headache and liver diseases. There are reports indicating various activities of this plant such as anti inflammatory, antimicrobial, free radical scavenging, antidepressant and anti allergic characteristics. We used ACMS (Accelerated cytotoxic or protective mechanism screening techniques) to evaluate possible antioxidant effect of Eremostachys laciniata against oxidative cell damage induced by different system of oxidative stress such as quinolin- iron complex and cupper in freshly isolated liver cell model.MATERIALS AND METHODSThe extracts of the plant material were prepared, with n-hexane, dichloromethane and methanol (1L each). The resulting combined extracts were dried using rotary evaporator. Hepatocytes were isolated from male Sprague-Dawley rats by a two-step collagenase perfusion, as described previously. Cell viability was measured by trypan blue exclusion method. The hepatocytes used in this study were at least 85-90% viable immediately after isolation. In the present study, five different assays were used to evaluate the antioxidant activity of the extract of Eremostachys laciniata growing in Iran DPPH (2,2-diphenyl-1-picrylhydrazyl) assay, the cell viability determined by trypan blue exclusion, ROS formation measured by using DCF-DA (2,7-dichlorofluorescin diacetate) prob, Mitochondrial membrane potential (MMP) was assessed by rhodamine 123, and lipid peroxidation assessed by TBA and TCA. RESULTSThe methanolic extract significantly decreased cell death and ROS formation induced by iron-quinoline complex and cupper as measured by trypan blue exclusion method and dichlorofluorescin diacetate fluorescence assay respectively. The methanolic extract of Eremostachys laciniata showed protective effect against Mitochondrial membrane potential collapse and lipid peroxidation caused by cupper. The methanolic extract were shown to possesses a significant 2, 2-diphenyl -1-picrylhydrazyl radical scavenging activity (RC50%=0.212). The n-hexane and dichloromethane extracts showed toxic effects.DISCUSSIONOur findings are in accordance with previous studies showing that oxidative stress plays a major role in iron-Quinoline and cupper cytotoxicity. Our methanolic extract showed protective effect and n-hexane and dichloromethane extracts showed toxic effect when added to cell suspensions. This protection can be attributed to antioxidant characteristics of our methanolic extract. Mechanisms such as chelating iron released from complex and free radical scavenging signaling pathways leading to MPT and necroses or apoptosis could be involved. Further investigation on isolated constituents is needed for clarification of the exact chemicals responsible for the protective and toxic effects of the extracts.