Chinese Journal of Dermatology 2015;(8):568-571
doi:10.3760/cma.j.issn.0412-4030.2015.08.012
Effects of alpha-lipoic acid on autophagy in human skin fibroblasts
Yunpeng ZHENG ; Xu CHEN ; Dan HUANG ; Song XU ; Heng GU
Keywords
Autophagy; Fibroblast; Cell proliferation; Microtubule-associated protein 1 light chain-3B
Country
China
Language
Chinese
Abstract
Objective 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.
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