蛋白激酶C在肝细胞缺氧预处理中的作用

Effect of protein kinase C during hepatocyte hypoxic precondition

目的:研究蛋白激酶C(PKC)在肝细胞缺氧预处理中的作用, 方法:建立体外肝细胞缺氧预处理模型,应用PKC抑制剂白屈菜季铵碱(chelerythrine chloride,CHE)和激动剂豆蔻酸佛波酰乙酯(phorobol12-myristate13-acetate,PMA),通过检测PKC磷酸化活性,细胞存活率,同时在透射电镜下观察肝细胞超微结构改变,研究PKC的作用,对相关数据进行统计学处理。结果:和缺氧复氧组的PKC磷酸化活性(710.5±78.8)fkat/g比较,缺氧预处理组的PKC磷酸化活性(1823.7±2682)fkat/g和PKC激动剂组的PKC磷酸化活性(2541.2±326.5)fkat/g显著增高(P<0.01),肝细胞结构损伤改变较小;和缺氧预处理组比较,PKC抑制剂组相应指标呈相反的变化,PKC磷酸化活性(1088.0±89.3)fkat/g(P<0.01)。结论:肝细胞缺氧预处理细胞保护作用中,PKC通路起到至关重要的作用。

AIM: To investigate the effects of protein kinase C (PKC) on hypoxic preconditioning (HP) for hepatocyte. METHODS: Through a normal liver cell HP model, PKC inhibitor and activator were utilized to analyze the phosphorylation of PKC. The cellular structure and viability were also observed. All the data were statistically analyzed. RESULTS: Compared with the phosphorylation of PKC in the control without HP[(710.5±78.8) fkat/g], the phosphorylation of PKC was obviously increased in HP treated model [(1 823.7±268.2) fkat/g] and PMA treated model[(2 541.2±326.5) fkat/g] (P<0.01). Cellular changes were less. In addition, opposite changes were found in PKC inhibited groups, and the phosphorylation of PKC was [(1 088.0+89.3) fkat/g] (P <0.01). CONCLUSION: The activation of PKC is the important chain of HP in the preservation of liver cell, and its mechanism may be involved in protein phosphorylation.