In vitro effects of asiaticoside on the proliferation of and connective tissue growth factor expression by keloid-derived fibroblasts / 中华皮肤科杂志

ABSTRACT

[Objective] To investigate the m vitro effect of asiaticoside on the proliferation of and connective tissue growth factor (CTGF) expression by keloid-derived fibroblasts.[Methods] Tissue samples from patients with keloid were obtained for primary culture of fibroblasts.After 3 to 7 passages,the fibroblasts were incubated with different concentrations (100,10,1,0.1,0.01 mg/L) of asiaticoside or dimethyl sulfoxide (DMSO) for 24,48 and 72 hours followed by the determination of cell viability by methyl thiazolyl tetrazolium (MTT) assay.Immunohistochemistry and Western blot were carried out to quantify the expression of CTGF in the fibroblasts at 48 hours after treatment with different concentrations of asiaticoside.The morphology of fibroblasts was observed before and after the treatment with asiatieoside.[Results] As morphological observation showed,different concentrations of asiaticoside induced an obvious apoptosis and growth inhibition in fibroblasts.The growth of fibroblasts was suppressed by asiaticoside of 1-100 mg/L in a dose-dependent manner (r =0.95,0.90,0.92 for 24-,48- and 72-hour treatment respectively,all P ＜ 0.01 ),and one-factor analysis of variance revealed statistical differences in the growth inhibition rate in fibroblasts between different treatment durations for each tested concentration of asiaticoside (all P ＜ 0.01 ).There was a strong expression of CTGF in untreated fibroblasts,which was weakened by the treatment with asiaticoside for 48 hours.The number of CTGF-positive fibroblasts per 100 cells was 73 in untreated fibroblasts,significantly higher than that in fibroblasts treated with asiaticoside at 1 mg/L (54,t =4.34,P ＜ 0.01 ) and 10 mg/L (46,t =6.26,P ＜ 0.01 ),and statistical differences were observed between the fibroblasts treated with asiaticoside at 1 mg/L and 10 mg/L (t =1.95,P ＜ 0.05).Western blot also showed that the expression of CTGF was inhibited by 48-hour treatment with asiaticoside,and the inhibitive effect displayed a trend to increase with the rise in the concentration of asiaticoside.[Conclusion] Asiaticoside can effectively inhibit the oroliferation of and CTGF expression by fibroblasts in vitro.