摘要目的：评价α硫辛酸对人皮肤成纤维细胞自噬水平的影响。方法人皮肤成纤维细胞与终浓度分别为0、0.05、0.10、0.15、0.20和0.50 mmol/L 的α硫辛酸孵育4、12和24 h 后，采用噻唑蓝法检测细胞增殖活性，单丹酰戊二胺（MDC）染色法检测细胞自噬水平，Western 印迹法检测微管相关蛋白1轻链3-B（LC3-B）表达。结果孵育4 h 和12 h 时，0、0.05、0.10、0.15、0.20和0.50 mmol/L α硫辛酸组间人皮肤成纤维细胞增殖活性（A490值）差异均无统计学意义（F 值分别为2.85、1.34，均 P >0.05）；孵育24 h 时，各组间差异有统计学意义（F =10.41，P <0.01）。MDC 法检测显示，孵育4 h 和12 h 时，0、0.05、0.10、0.15、0.20和0.50 mmol/L α硫辛酸组间自噬体阳性细胞百分率差异均无统计学意义（F 值分别为0.11、0.10，均 P >0.05）；孵育24 h 时，各组间差异有统计学意义（F =8.03，P <0.05）。Western 印迹法检测显示，孵育4 h 和12 h 时，0、0.05、0.10、0.15、0.20和0.50 mmol/L α硫辛酸组间细胞 LC3-Ⅰ向 LC3-Ⅱ转化（LC3-Ⅱ/LC3-Ⅰ）水平差异均无统计学意义（F 值分别为3.38、2.13，均 P >0.05），但孵育24 h 时各组间差异有统计学意义（F =37.49，P <0.01）。结论α硫辛酸可能抑制人皮肤成纤维细胞基础自噬。

abstractsObjective To evaluate the effects of alpha-lipoic acid (α-LA)on autophagy in human skin fibroblasts (HSFs). Methods HSFs at passage 3 - 5 were divided into several groups to be cultured with α-LA at final concentrations of 0, 0.01, 0.05, 0.10, 0.15, 0.20 and 0.50 mmol/L for 4, 12 and 24 hours, respectively. Methyl thiazolyl tetrazolium(MTT)assay was performed to evaluate cellular proliferative activity, monodansylcadaverin(MDC)staining to determine autophagy levels, and Western blot to measure the expression of the microtubule-associated protein 1 light chain-3B(LC3-B). Results After incubation for 24 hours, there was a significant difference in the proliferative activity of HSFs among all the groups (F = 10.41, P < 0.05), while no significant differences were observed after incubation for either 4 or 12 hours (F = 2.85, 1.34, respectively, both P > 0.05). MDC staining also showed a significant difference in the percentage of autophagosome-positive cells among all the groups after 24-hour incubation (F = 8.03, P < 0.05), but no significant difference after either 4- or 12-hour incubation (F = 0.11, 0.10, respectively, both P > 0.05). Western blot revealed that the degree of conversion from LC3-Ⅰ to LC3-Ⅱ(LC3-Ⅱ/LC3-Ⅰratio)was significantly different among all the groups after 24-hour incubation (F = 37.49, P < 0.05), but similar after 4- and 12-hour incubation (F = 3.38, 2.13, respectively, both P > 0.05). Conclusion α-LA may inhibit basal autophagy in HSFs.