LPS致肝细胞损伤模型的建立研究

To Establish and Research Liver Cell Injury Model Induced by LPS

[目的]探索LPS体外致人Chang Liver细胞损伤模型的建立方法和意义。[方法]分别用20、40、60、80、100μg/ml浓度的LPS作用Chang Liver细胞24 h,采用MTT法检测LPS对Chang Liver细胞存活率的影响,分别测定Chang Liver细胞上清中谷草转氨酶(AST)、谷丙转氨酶(ALT)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(γ-GT)、乳酸脱氢酶(LDH)的活性,并通过Hoechst 33258荧光染色观察用药24 h后Chang Liver细胞形态的变化。[结果]80和100μg/ml浓度的LPS作用后Chang Liver细胞存活率显著性降低,胞外AST、ALT、ALP、γ-GT、LDH酶活性升高,细胞数目减少,细胞形态发生改变,呈现细胞核固缩等细胞凋亡现象。[结论]用80μg/ml的LPS与Chang Liver细胞共培养24 h,可以建立理想的体外肝细胞损伤模型。

[ Objective ] To establish in vitro LPS-induced Chang Liver cells damage model [ Method] After received LPS 24h at different con- centrations （20, 40, 60, 80, 100 μg/ml）, Chang Liver cells viability effects were detected by MTT methods, and the AST, ALT, ALP,γ-GT, LDH activity in ceU supernatant were measured. The Chang Liver cells morphology changes were showed by Hoechst 33258 staining. [ Result] Chang Liver cell viability was significantly reduced with LPS in 80 and 100 μg/ml. AST, ALT, ALP, γ-GT, LDH activity in cell supernatant were increased; the number of ceils were reduced; ceU morphology was changed showing cell apoptosis with nuclear condensation. [ Conclusion ] Chang Liver ceils co-cultured with 80μg/ml LPS might be an ideal in vitro model of hepatocellular injury.