利多卡因对缺氧/复氧后肝细胞钙超载和细胞凋亡的影响

Effect of lidocaine precondition on hepatocytes calcium overload and apoptosis induced by cell hypoxia-reoxygenation

目的:探讨利多卡因预处理对L02肝细胞缺氧/复氧后肝细胞内钙离子变化和细胞凋亡的影响。方法:选用L02肝细胞缺氧/复氧损伤模型,随机分为缺氧/复氧组(I组)、利多卡因预处理组(II组)和正常对照组(III组);检测肝细胞缺氧4h,复氧后10h细胞培养基中谷丙转氨酶(ALT)浓度、谷草转氨酶(AST)浓度、肝细胞内钙离子浓度、肝细胞凋亡率以及细胞形态结构变化。结果:缺氧/复氧后肝细胞内钙离子浓度升高,且与细胞凋亡率呈正相关(P<0.05);I,II组细胞复氧10h后肝细胞培养液中ALT浓度、AST浓度、肝细胞内钙离子浓度、细胞凋亡率较III组均升高(P<0.05),倒置显微镜和电子显微镜示肝细胞损伤,可见凋亡细胞;II组细胞复氧10h后肝细胞培养液中ALT浓度、AST浓度、肝细胞内钙离子浓度、细胞凋亡率均低于I组(P<0.05),倒置显微镜和电子显微镜结果也显示II组肝细胞比I组损伤轻微。结论:利多卡因预处理可以缓解缺氧/复氧损伤后肝细胞内钙超载,降低细胞凋亡率。

Objective To investigate the effect and possible mechanism of lidocaine precondition on calcium overload and apoptosis of the hepatocytes induced by cell hypoxia-reoxygenation. Methods The cultured L02 hepatocytes were randomly divided into 3 groups： a hypoxia-reoxygenation group （Group Ⅰ ） , a lidocaine precondition group （Group Ⅱ ） , and a normal control group （ Group Ⅲ ）. After 4 hours of cell hypoxia and 10 hours of reoxygenation, alamine aminotransferase （ALT） and aspartate aminotransferase （AST） concentrations in the nutritive medium were detected. The cytoplasm ionic calcium concentration was measured by fluorospectrophotometer. The apoptosis was measured by flow cytometry and fluorescent microscope. The appearance and ultramicrostructure changes of hepatocytes were observed by inverted microscope and electronic microscope. Results Cytoplasm ionic calcium concentration and apoptosis was positively correlated （ r = 0. 7652, R^2 = 0. 5855, P 〈 0.05 ）. The ALT concentration in the nutritive medium, AST concentration in the nutritive medium, cytoplasm ionic calcium concentration and the ratio of apoptosis of Group Ⅰ and Ⅱ were significantly higher than those of Group Ⅲ（ P 〈 0.05 ）. The appearance and uhramicrostructure changes of Group Ⅰ and Ⅱ were worse than those of Group Ⅲ. The ALT concentration in the nutritive medium, AST concentration in the nutritive medium, cytoplasm ionic calcium concentration and the ratio of apoptosis of Group Ⅱ were significantly lower than those of Group Ⅰ （ P 〈 0.05 ）. The ultramicrostructure injury of hepatocytes of Group Ⅱ were less serious than those of Group Ⅰ. Conclusion Precondition with lidocaine can attenuate calcium overload of hepatocytes induced by hypoxia-reoxygenation in vitro, and decrease the ratio of apoptosis.