槲皮素对酒精性肝损伤的防护作用及其与I型血红素氧化酶关系的研究

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。

THE PREVENTIVE EFFECTS OF QUERCETIN ON ETHANOL-INDUCED HEPATOCYTES DAMAGE AND THE POSSIBLE MECHANISM INVOLVING HEME OXYGENASE-1

Objective To investigate the preventive effects of quercetin on ethanol-induced oxidative damage in rat primary hepatocytes and its relation with heme oxygenase-1 (HO-1). Method Sprague- Dawley rat primary hepatocytes were isolated using two-step collagenase perfusion technique. The ethanol-intoxicated rat hepatocytes were respectively post-treated (2h), simultaneously treated or pre-treated (2 h) with quercetin (50 μmol/L) in presence or absence of zinc protoporphyrin Ⅸ(ZnPP Ⅸ), an HO-1 inhibitor. Lactate dehydrogenase (LDH), aspartate transaminase (AST), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) were determined to evaluate the preventive effects. Results 50 μmol/L quercetin pre-treatment 2 h before ethanol insult could significantly prohibit the release of AST and LDH, the rise of MDA level and the decline of SOD and CAT activities and GSH content of hepatocytes exposed to ethanol. With addition of ZnPPⅨ , the protection of quercetin was partly reversed. There were significant differences of LDH, AST, MDA, GSH, SOD and CAT levels in rat hepatocytes compared with quercetin pre-treatment only. Conclusion The toxic insult of ethanol to rat hepatocytes could be challenged by quercetin pretreatment, and HO-1 was probably involved in the protection mechanism.