论文部分内容阅读
目的:探讨补青颗粒含药血清对高糖诱导的人晶状体上皮(HLE)细胞线粒体自噬的影响。方法:体外培养HLE细胞,空白组将HLE细胞在正常培养液中培养48 h,模型组在空白组培养液中添加30 mmol·L~(-1)葡萄糖,实验组在模型组基础上加入不同含药血清刺激培养48 h。采用CCK-8法检测不同含药血清对高糖诱导的HLE细胞增殖活力的影响;利用免疫荧光技术及蛋白免疫印迹(Western blot)法检测Pink-1,Parkin,p62蛋白表达。结果:CCK-8法结果显示模型组较空白组HLE细胞增殖活力增强(P<0.01);与模型组比较,杞菊组与补青颗粒高、中、低剂量组细胞增殖活力均降低(P<0.01);与杞菊组比较,补青颗粒中剂量组细胞增殖活力明显降低(P<0.01);补青颗粒不同剂量组间比较,补青颗粒中剂量组细胞增殖活力显著降低(P<0.01)。细胞免疫荧光结果显示Pink-1,Parkin,p62蛋白主要在细胞胞浆表达,细胞核内亦有少量表达。Western blot结果显示与空白组比较,模型组Pink-1,Parkin表达增加(P<0.01),p62表达降低(P<0.01);与模型组比较,杞菊组与补青颗粒中剂量组Pink-1,Parkin表达增加(P<0.01),p62表达降低(P<0.01);与杞菊组比较,补青颗粒中剂量组Pink-1,Parkin表达增加(P<0.01,P<0.05),p62表达降低(P<0.01)。结论:补青颗粒对高糖诱导的HLE细胞线粒体自噬相关蛋白的表达有显著影响,线粒体自噬的改变可能是补青颗粒改善白内障发病的机制之一。
Objective: To investigate the effect of Buqing granule-containing serum on mitophagy of human lens epithelial (HLE) cells induced by high glucose. Methods: HLE cells were cultured in vitro. HLE cells were cultured in normal medium for 48 h in the blank group, and 30 mmol·L -1 glucose in the blank group were added to the model group. The experimental group was added to the model group Drug-containing serum stimulated for 48 h. The CCK-8 method was used to detect the effect of different drug-containing serum on the proliferation of HLE cells induced by high glucose. The protein expressions of Pink-1, Parkin and p62 were detected by immunofluorescence and Western blot. Results: The results of CCK-8 showed that the proliferation activity of HLE cells in model group was higher than that in blank group (P <0.01). Compared with model group, the cell proliferation activities of QJ and CHQ groups were lower (P < 0.01) .Compared with QIJU group, the cell proliferation activity of middle-dose group was significantly decreased (P <0.01); Compared with QQ group, the cell proliferation activity of middle-dose group was significantly decreased (P <0.01) . Immunofluorescence results showed that the expression of Pink-1, Parkin and p62 protein was mainly in the cytoplasm of the cells, but also in the nucleus. Western blot results showed that compared with the blank group, the expressions of Pink-1 and Parkin in the model group increased (P <0.01) and the expression of p62 decreased (P <0.01). Compared with the model group, (P <0.01). Compared with Qijuzhi group, the expressions of Pink-1 and Parkin increased (P <0.01, P <0.05) and the expression of p62 decreased (P <0.01). CONCLUSION: BuQing granule has a significant effect on the expression of mitochondria autophagy-related protein in HLE cells induced by high glucose. The change of mitochondrial autophagy may be one of the mechanisms of BuQing granules in improving the incidence of cataract.