Objective In order to explore the inhibitory effects of cedrol on the activation of androgen receptor(AR)in cultured mouse embryonic dermal fibroblasts(meFBs)treated with dihydrotestosterone.Methods Primary cultures of meFBs were established from the embryonic day 18.5(E18.5)C57BL/6J mouse skin tissues.The aggregative growth pattern of the meFBs was observed by an inverted microscope.The activity of alkaline phosphatase(ALP)and CD 133 expression were examined using an ALP assay kit and fluorescence staining technology.The exponentially growing meFBs were divided into four groups,including the control group,the dihydrotestosterone(DHT)group,the cedrol group,and the combination group with cedrol and DHT.Cell counting kit-8(CCK-8)was used to determine cell viability of the cells treated with varying concentrations(0-300 mol/L)of cedrol and DHT for 48 h.The expression and distribution of AR and AR coactivator four and a half LIM domains 2(FHL2)were also examined using immunofluorescence staining,western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)technique,respectively.The ALP activity was detected in the treated cells as well.Results The intense staining of AR was observed in the nuclei of meFBs treated with 30 μmol/L DHT for 48 h,the nucleus-to-cytoplasm ratio of AR immunofluorescent intensity was 3.39±0.04 per 10 cells in the DHT-treated cells and was 1.24±0.02 per 10 cells in the cells treated with a combination of cedrol and DHT.There were significant differences between DHT group and DHT+cedrol group(P＜0.05).The authors also observed that DHT could down-regulate the expression of ALP and up-regulate the expression of AR co-activating factor FHL2,while cedrol could reverse the effect of DHT for fibroblast.Conclusion Cedrol has a potent anti-androgen effect on in vitro cultured meFBs by preventing DHT-induced AR nuclear translocation and by inhibiting the expression of AR-coactivator FHL2.Topical use of cedrol might be a promising treatment for androgenetic alopecia,which is worthy of further investigation.
维普
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