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为分析NADH在保护正常肝细胞氰化物缺氧性损伤中的作用 ,在L0 2细胞组织培养基中加入 3mmol/LKCN后立即加入 0～6 0 0 μg/ml的NADH ,培养 0 .5、2、4h后检测细胞凋亡率和Bcl 2家族蛋白的表达。另取L0 2细胞分 3组进行实验 ,实验组Ⅰ为对照组 ,实验组Ⅱ、Ⅲ加 3mmol/LKCN后分别加入不含和含有NADH(40 0 μg/ml)的RPMI 16 4 0培养 2h,检测 3组Bcl XL ,Bax,Bcl 2蛋白表达 ,并取样进行透射电镜观察细胞形态。结果显示 ,NADH对氰化细胞凋亡率的抑制作用呈剂量依赖性 ,在浓度为 4 0 0～ 6 0 0 μg/ml时达平台 ,并且能够上调Bcl 2及Bcl XL蛋白表达 ,下调Bax蛋白表达 ;电镜观察可见损伤后细胞呈凋亡和坏死改变。提示NADH具有明显抗肝细胞氰化物损伤作用 ,其作用机制可能与Bcl 2、Bcl XL蛋白表达的上调及和Bax蛋白表达的下调有关。
In order to analyze the role of NADH in protecting normal liver cells from hypoxia injury of cyanide, 0-600 μg / ml NADH was added immediately after adding 3 mmol / L KCN into L0 2 cell culture medium, After 4h, the apoptosis rate and the expression of Bcl2family protein were detected. In addition, L0 2 cells were divided into 3 groups for experiment. The experimental group Ⅰ was the control group. The experimental group Ⅱ and Ⅲ plus 3mmol / LKCN were cultured in RPMI 1640 without and with NADH (40 0 μg / ml) The expressions of Bcl-XL, Bax and Bcl-2 protein in the three groups were detected and the morphological changes were observed by transmission electron microscope. The results showed that the inhibitory effect of NADH on cyanide cell apoptosis in a dose-dependent manner, at a concentration of 400 ~ 600 μg / ml up to the platform, and can increase Bcl 2 and Bcl XL protein expression, down Bax protein The changes of apoptosis and necrosis of cells were observed under electron microscope. These results suggest that NADH has obvious anti-hepatocyte cytotoxic effect, which may be related to the up-regulation of Bcl-2 and Bcl-XL protein expression and the down-regulation of Bax protein expression.