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目的研究槲皮素对大鼠原代肝细胞酒精性氧化损伤的防护作用及其与I型血红素氧化酶(HO-1)的关系。方法二步胶原酶技术分离培养大鼠原代肝细胞。分别在酒精染毒前、后及同时用50μmol/L槲皮素作用,作用结束后检测细胞培养上清液中乳酸脱氢酶(LDH)、谷草转氨酶(AST)活性,细胞丙二醛(MDA)、谷胱甘肽(GSH)水平和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性;槲皮素预作用同时加入HO-1抑制剂锌原卟啉Ⅸ(ZnPPⅨ),作用结束后测上述指标。结果酒精暴露前2 h经50μmol/L槲皮素干预,肝细胞LDH、AST和MDA水平较酒精组明显降低,GSH、SOD和CAT水平较酒精组明显升高;槲皮素预干预同时加ZnPPⅨ可抑制HO-1活性,LDH、AST和MDA水平较未加有明显升高,而GSH、SOD和CAT水平则明显降低。结论槲皮素预干预对肝细胞酒精性氧化损伤具有防护作用,其保护效应可能通过HO-1介导。
Objective To study the protective effect of quercetin on alcoholic oxidative damage of rat primary hepatocytes and its relationship with type I heme oxygenase (HO-1). Methods Two - step collagenase technique was used to isolate rat primary hepatocytes. The cells were treated with 50μmol / L quercetin before, after and simultaneously with alcohol exposure. The activities of lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and malondialdehyde (MDA) ), Glutathione (GSH) and superoxide dismutase (SOD) and catalase (CAT) activity were observed. Pretreatment with quercetin was simultaneously added ZnPPⅨ, a HO-1 inhibitor, After the end of the role of the above indicators. Results The levels of LDH, AST and MDA in hepatocytes were significantly lower than those in the alcohol group and the levels of GSH, SOD and CAT were significantly increased 2 h before alcohol exposure with 50 μmol / L quercetin. Compared with the alcohol group, quercetin pretreatment added ZnPPⅨ Inhibited the activity of HO-1, the levels of LDH, AST and MDA were significantly increased compared with no increase, while the levels of GSH, SOD and CAT were significantly decreased. Conclusion Pretreatment with quercetin has a protective effect on alcoholic oxidative damage of hepatocytes, and its protective effect may be mediated by HO-1.