西罗莫司对人皮肤成纤维细胞自噬水平的影响

Effects of sirolimus on the autophagy in human skin fibroblasts

目的研究西罗莫司对人皮肤成纤维细胞自噬水平的影响。方法取健康青少年男子环切术后包皮进行人皮肤成纤维细胞的分离培养,西罗莫司浓度分别为0、0.1、1.0、10.0 mg/L处理24 h,并设立空白对照。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beclin1的表达水平。数据采用SPSS16.0软件分析,多组间均数行单因素方差分析进行比较。结果 0、0.1、1.0、10.0 mg/L西罗莫司组吖啶橙染色荧光定量呈浓度依赖性升高,各组间差异均有统计学意义(P<0.01);LC3-B及beclin 1的表达水平明显升高,各组间差异均有统计学意义(P<0.01)。结论西罗莫司可以提高人皮肤成纤维细胞的自噬水平。

Objective To investigate the effects of sirolimus on the autophagy in human skin fibroblasts（HSFs）. Methods HSFs were isolated from the circumcised foreskin of a healthy male, and subjected to primary culture. After 4 to 10 passages, HSFs were incubated with sirolimus of 0, 0.1, 1.0, 10.0 mg/L for 24 h followed by the observation of autophagy. Aendine orange（AO） staining under florescent microscope and transmission election microscope to detect autophagy, Western blot to detect the expressions of LC3-B and beelinl in these skin fibroblasts. Results Intracellular acridine orange accumulation increased, and significant differences were observed between the 4 groups, beclinl and LC3-B expression were up-regulated significantly compared to those untreated skin fibroblasts （all P〈0.01）. Conclusion The autophagy in HSFs can be upregulated by sirolimus.