沉默miR-23a对中波紫外线诱导成纤维细胞自噬的作用研究

Effect of miR-23a Silencing on Ultraviolet B-suppressed Depression of Autophagy Flux to Fibroblasts

目的探讨沉默mi R-23a对中波紫外线(UVB)诱导成纤维细胞自噬的作用。方法将成纤维细胞分为空白对照组、UVB光老化组、mi R-23a沉默组、UVB光老化+mi R-23a沉默组。吖啶橙染色检测细胞自噬,Western印迹检测自噬相关蛋白LC3-B及p62的表达水平。结果空白对照组与UVB光老化组吖啶橙染色荧光定量分别为(39.34±1.15)%、(32.22±1.40)%,2组间差异有统计学意义(P<0.05);UVB光老化+mi R-23a沉默组吖啶橙染色荧光定量为(43.72±0.95)%,与UVB光老化组相比,差异有统计学意义(P<0.05)。与空白对照组相比,UVB光老化组自噬相关蛋白p62表达量升高,LC3-Ⅰ转化为LC3-Ⅱ减少,2组间差异有统计学意义(P<0.05);与UVB光老化组相比,UVB光老化+mi R-23a沉默组p62表达量下降,LC3-Ⅰ转化为LC3-Ⅱ增多,2组间差异有统计学意义(P<0.05)。结论 UVB辐射可致成纤维细胞自噬水平下降,而沉默mi R-23a可抑制UVB诱导所致的自噬水平下降。

Objective To explore effects of the expression of silent miRNA-23 a on UVB irradiation-induced suppression of autophagy flux of fibroblasts. Methods Fibroblasts were divided into four groups including normal control group, UVB irradiation group, miR-23 a silencing group and UVB irradiation with miR-23 a silencing group. Autophagy level was determined by acridine orange staining followed by fluorescence microscopy and transmission electron microscopy. Autophagy related p62 protein and LC3 were detected by western blotting. Results Compared to normal control group, the intracellular intensity of acridine orange induced fluorescence and the protein level of LC3- Ⅱ was down-regulated and the p62 protein expression was up-regulated for the UVB irradiation group. Significant differences were also observed between the UVB irradiated with miR-23 a silencing group and UVB irradiation group. Conclusion Silencing the expression of miR-23 a can inhibit UVB-induced suppression of autophagy flux.