虎杖苷对非酒精性脂肪性肝细胞内质网应激的调控机制

Regulatory mechanism of Polygonidine on endoplasmic reticulum stress in non-alcoholic fatty hepatocytes

目的 探讨虎杖苷对小鼠非酒精性脂肪肝疾病中肝脏细胞内质网应激、细胞自噬和凋亡的影响。方法 采用高脂饮食诱导小鼠NAFLD疾病模型,对模型小鼠体质量进行监测,在造模结束后检测TG、LDL和ALT等生化指标。体外培养Huh7细胞,用PA诱导细胞损伤。用Annexin V/PI双染法检测细胞凋亡,用蛋白质免疫印迹实验检测内质网应激标记分子Bip、p-eIF2α、CHOP、ATF6和p-IRE1的表达和自噬标记分子LC3和p62的表达;采用免疫荧光染色法检测LC3蛋白的定位。结果 与对照组相比,灌胃小鼠高脂饮食条件下的体质量、TG和LDL显著下降(P<0.05);虎杖苷处理组HepG2细胞PA诱导的细胞凋亡水平显著下降(P<0.05);内质网应激标记分子Bip、p-eIF2α、CHOP、ATF6和p-IRE1的表达水平显著下降(P<0.05);细胞自噬活性明显增加。结论 虎杖苷通过抑制内质网应激和细胞自噬保护高脂诱导的肝损伤。

Objective To investigate the effects of polygonidine on endoplasmic reticulum stress,autophagy and apoptosis of liver cells in mice with non-alcoholic fatty liver disease. Methods The mouse model of NAFLD was induced by high-fat diet. The body weight of the model mice was monitored,and the biochemical indexes such as TG,LDL and ALT were detected after the modeling. Huh7 cells were cultured in vitro and induced by PA. Annexin V/PI staining was used to detect cell apoptosis. Western blotting was used to detect the expression of endoplasmic reticulum stress markers Bip,P-EIF2α,CHOP,ATF6 and P-IRE1,and the expression of autophagy markers LC3and P62. The localization of LC3 protein was detected by immunofluorescence staining. Results Compared with the control group,the body weight,TG and LDL of mice treated with high-fat diet were significantly decreased(P<0.05). The PA-induced apoptosis level of HepG2 cells in PD treated group was significantly decreased(P<0.05).The expressions of endoplasmic reticulum stress markers Bip,P-EIF2α,CHOP,ATF6 and P-IRE1 were significantly decreased(P<0.05). Autophagy activity was significantly increased. Conclusion Polygonin protects against liver injury induced by high fat by inhibiting ER stress and autophagy.