二咖啡酰奎宁酸对大鼠肝原代细胞保护作用的研究

Protective effects of Dicaffeoylguinic Acid against CCl_4-induced injury in primary cultured rat hepatocytes ①

目的 采用肝原代细胞培养及四氯化碳损伤法 ,观察 1 ,5 二 O 二咖啡酰奎宁酸对肝细胞的保护作用 ,并通过体外实验初步探讨二咖啡酰奎宁酸保护肝细胞的作用机制。方法 采用肝细胞消化酶两步灌注法 ,肝细胞消化液消化肝细胞 ,梯度离心分离肝细胞 ,细胞获得量为 (1 - 1 0 )× 1 0 7·ml-1 ,肝细胞活力在 90 %以上 ,以此为靶细胞。并利用四氯化碳损伤方法制造肝细胞损伤模型。分别采用3 H -胸腺嘧啶 (3 H -TdR) ,3 H -脯氨酸 (3 H -Pro)掺入 ,流式细胞术等方法 ,观察二咖啡酰奎宁酸对肝细胞的增殖、活力、凋亡及细胞周期等的影响 ,并测定天门冬氨酸转移酶活性。结果 二咖啡酰奎宁酸在 7.8～ 2 5 0mg·L-1浓度对肝细胞无明显毒性作用 ,能促进正常及四氯化碳损伤的肝细胞增殖及增强细胞活力 ,降低细胞上清的天门冬氨酸转移酶 (AST)活性 ,抑制四氯化碳引起的肝细胞凋亡以及改变受损细胞周期。结论 二咖啡酰奎宁酸具有显著的肝细胞保护作用 ,其作用机制为促进肝细胞的增殖及活力 ,抑制受损肝细胞的凋亡 。

Aim To study the effects of dicaffeoylguinic acid on carbon tetrachloride (CCl 4) induced primary cultured rat hepatocytes.Methods Rat hepatocytes were isolated and injured by CCl 4.The proliferation and the viability of the hepatocytes were assayed by measuring the incoporating 3 H TdR and 3 H Pro respectively.The apoptosis and cell cycle of hepatocyte were determined by flow cytometry (FCM).And the activity of AST was examined.Results Dicaffeoylguinic acid 7.8～250mg·L -1 had no toxicity to the hepatocytes. The compound markedly increased the proliferation and the viability of the normal and injured rat hepatocyte,decreased AST activity in the cultural medium; restrained the hepatocytes apoptosis by CCl 4 and changed the cell cycle of hepatocytes.Conclusion Dicaffeoylguinic acid has notable anti-hepatocytes-injury effects in vitro.