肝宁方对衣霉素诱导肝细胞内质网应激生存信号分子的影响

Effect of the Ganning Formula on the Survival Signal Molecules Induced by Tunicamycin Under Endoplasmic Reticulum Stress

目的研究肝宁方对衣霉素诱导的人肝细胞内质网应激条件下的生存信号分子的影响。方法将肝细胞分正常组、模型组、治疗组:正常组给予正常大鼠血清培养基培养48h,模型组在正常组基础上给予衣霉素诱导48h,治疗组在模型组基础上给予肝宁方继续培养24h。Western blot法检测各组蛋白ATF6、IRE1α、PERK、GRP78的表达量,分析蛋白表达差异。结果肝宁方治疗组与衣霉素诱导模型组比较,治疗组内质网应激信号分子的蛋白ATF6、IRE1α、PERK的表达明显减少(P<0.05);模型组与对照组比较,模型组GRP78、内质网应激信号分子的蛋白ATF6、IRE1α、PERK的表达明显增多(P<0.05)。结论衣霉素诱导正常肝细胞后可激活ATF6、IRE1α、PERK及GRP78蛋白的表达,肝宁方对衣霉素诱导内质网应激生存信号分子蛋白ATF6、IRE1α、PERK的表达及内质网应激标记蛋白均有抑制作用,从而达到阻止未折叠蛋白的错误激活、抑制肝细胞凋亡,对肝细胞具有保护作用。

Objective To study the effect of Ganning Formula on the survival signal molecules of human hepatocytes induced by tunicamycin under endoplasmic reticulum stress.Methods Liver cells were divided into normal group, model group and treatment group. The normal group was incubated with normal rat serum for 48h. The model group treated with tunicamycin for 48h on the basis of normal group. The treatment group were treated with Ganning Formula for 24h on the basis of model group. The protein expression levels of ATF6, IRE1α, PERK and GRP78 were detected by Western blot. And the differences in protein expression were analyzed by statistical software SPSS18.0.Results Compared with the tunicamycin-induced model group, the expression of ATF6, IRE1α and PERK on the endoplasmic reticulum stress signaling molecule were significantly decreased （P〈0.05） in the treatment group. Compared with the control group,the GRP78 and endoplasmic reticulum stress signal molecular ATF6, IRE1α, perk in the model group were increased significantly（P〈0.05）.Conclusion The expression of ATF6, IRE1α, PERK and GRP78 proteins was activated by tunicamycin-induced in normal hepatocytes. Ganning formula can inhibition expression of endoplasmic reticulum stress survival signal molecular protein ATF6, IRE1α, PERK and marker protein, which to prevent not protein folding error activation and inhibition of apoptosis of liver cells, has a protective effect on the liver cells.