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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 study the effect of sirolimus on the autophagy of human dermal fibroblasts. Methods Human dermal fibroblasts were excised from the circumcision of healthy young men. The concentrations of sirolimus were 0, 0.1, 1.0 and 10.0 mg / L, respectively, for 24 h. A blank control was established. Autophagy was detected by acridine orange staining. Western blotting was used to detect the expression of autophagy-related proteins LC3-B and beclin1. The data were analyzed by SPSS16.0 software, and multiple groups were compared by one-way ANOVA. Results Fluorescence quantification of acridine orange in 0, 0.1, 1.0, 10.0 mg / L sirolimus group showed a concentration-dependent manner, with significant difference between the groups (P <0.01); LC3-B and beclin 1 (P <0.01). There was significant difference between the two groups (P <0.01). Conclusion Sirolimus can increase autophagy in human dermal fibroblasts.