Effects of alpha-lipoic acid on autophagy in human skin fibroblasts
10.3760/cma.j.issn.0412-4030.2015.08.012
- VernacularTitle:α硫辛酸对人皮肤成纤维细胞自噬的影响
- Author:
Yunpeng ZHENG
;
Xu CHEN
;
Dan HUANG
;
Song XU
;
Heng GU
- Publication Type:Journal Article
- Keywords:
Autophagy;
Fibroblast;
Cell proliferation;
Microtubule-associated protein 1 light chain-3B
- From:
Chinese Journal of Dermatology
2015;(8):568-571
- CountryChina
- 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.
