体外大鼠肝细胞坏死性损伤模型的建立

Establishment of necrotic injured models of primary hepatocytes from rats

目的 建立体外肝细胞坏死性损伤模型,为进一步研究药物对体外肝细胞损伤的保护作用奠定基础。方法 采用Ⅳ型胶原酶灌流法分离大鼠肝实质细胞并进行体外培养,利用四氯化碳(CCl4)和过氧化氢(H2O2)体外诱导肝细胞坏死性损伤,检测ALT、AST、MDA、GSHpx等指标和采用MTT比色法。结果 CCl4体外诱导肝细胞损伤的最适损伤浓度和损伤时间为:8mmol·L-1,6h;H2O2体外诱导肝细胞坏死性损伤的最适损伤浓度和损伤时间为06mmol·L-1,1h。结论 利用最适损伤浓度的CCl4和H2O2分别与大鼠肝细胞共培养最适损伤时间,可建立两种理想的体外肝细胞坏死性损伤模型。

Objective To establish the nectotic injured models of hepatocytos in vitro for further studying protective effects of drugs on injured hepatocytes. Methods The hepatocytes were isolated by perfusion of liver with Ⅳ type collagenase in rats and incubated with CCl 4 or H 2O 2. The necrotic hepatocytes induced by CCl 4 or H 2O 2 were investigated by measuring the levels of AST and ALT, the amount of MDA and the activity of GSHpx, and MTT colormetry. Results The most optimal injured conditions of CCl 4 or H 2O 2 were separately determined as CCl 4: 8 mmol·L -1 , 6 h; H 2O 2: 0 6 mmol·L -1 , 1 h. Conclusion Two kinds of necrotic injured models of hepatocytes may be created separately by the most optimal injured conditions of CCl 4 or H 2O 2.