论文部分内容阅读
目的 从细胞凋亡的角度探讨阻塞性黄疸肝损害的调控机理。方法 采用胶原酶原位肝灌注法获取大鼠肝细胞 ,行原代培养 ,使用不同浓度甘氨鹅脱氧胆酸钠 (GCDC)及蛋白激酶C激动剂PMA、拮抗剂白屈菜赤碱 (chelerythrine)作用肝细胞后 ,用流式细胞术 (FCM)分析肝细胞的凋亡比率 ,用生物素 dUTP标记的TUNEL技术进行凋亡的原位检测。结果 随GCDC浓度的增加 ,肝细胞的凋亡比率明显增加 ,15 0 μMGCDC作用后肝细胞的凋亡率达到 (38 2 3± 1 5 8) % ;随PMA浓度的增加 ,肝细胞的凋亡明显增加 ,随白屈菜赤碱的增加 ,肝细胞的凋亡明显减少。结论 胆盐致肝细胞的凋亡 ,这可能是阻塞性黄疸肝损害的机理 ,蛋白激酶C信号通道起重要的调控作用。
Objective To investigate the regulatory mechanism of hepatic impairment in obstructive jaundice from the perspective of apoptosis. Methods Rat hepatic cells were obtained by collagenase in situ hepatic perfusion. Primary culture was performed. Glycyrrhizin deoxycholate (GCDC) and protein kinase C agonist (PMA) at different concentrations were used. The antagonist chelerythrine ) On hepatocytes, the rate of apoptosis in hepatocytes was analyzed by flow cytometry (FCM) and the in situ detection of apoptosis was performed using the biotin-dUTP-labeled TUNEL technique. Results With the increase of GCDC concentration, the apoptosis rate of hepatocytes increased significantly, and the apoptosis rate of hepatocytes reached to (38 2 3 ± 1 58)% after 15 μM GCDC treatment. With the increase of PMA concentration, the apoptosis of hepatocytes Significantly increased, with the increase of chelerythrine, hepatocyte apoptosis decreased significantly. Conclusion Bile salt induces apoptosis of hepatocytes, which may be the mechanism of hepatic injury in obstructive jaundice. Protein kinase C signaling plays an important regulatory role.