雪松醇抑制二氢睾酮诱导胎鼠真皮成纤维细胞雄激素受体活化  

Suppression of Cedrol on the Activation of Androgen Receptor in Cultured Mouse Embryonic Dermal Fibroblasts Treated with Dihydrotestosterone

作　　者：李潇 董炳琦 乐悦 廖志锴 雷铁池[1] Li Xiao;Dong Bingqi;Le Yue;Liao Zhikai;Lei Tiechi(Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学人民医院,湖北武汉430060

出　　处：《中国中西医结合皮肤性病学杂志》2024年第2期108-113,共6页Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金项目(编号:81972919)。

摘　　要：目的探究雪松醇对体外培养胎鼠真皮成纤维细胞二氢睾酮(DHT)诱导雄激素受体(AR)活化的抑制作用。方法分离第18.5天(E18.5)C57BL/6J胎鼠真皮,建立原代胎鼠真皮成纤维细胞培养;用倒置显微镜观察胎鼠成纤维细胞凝集生长;用碱性磷酸酶(ALP)试剂盒和荧光染色技术检测胎鼠成纤维细胞的ALP活性和CD133表达。将指数生长期胎鼠真皮成纤维细胞分为4组,即空白对照组、DHT组、雪松醇组和雪松醇+DHT联合处理组。不同浓度(0~300mol/L)雪松醇和DHT处理细胞48h后,用细胞计算试剂盒(CCK8)测定细胞存活率;用免疫荧光法、蛋白质印迹法(Westernblotting)和实时荧光定量聚合酶链反应(qRT-PCR)法检测各组AR及其共激活因子四个半LIM结构域2(FHL2)的表达水平和AR核定位;用ALP染色法检测各组细胞ALP活性变化。结果30μmol/L DHT处理胎鼠真皮成纤维细胞,48 h后观察到AR主要分布在细胞核,胞核/胞质荧光强度比值为(3.39±0.04)/高倍镜视野(HPF)。然而,雪松醇+DHT联合处理组胎鼠真皮成纤维细胞AR着色分布在胞浆和胞核,胞核/胞浆荧光强度比值为(1.24±0.02)/HPF,二者比较差异有统计学意义(P<0.05),提示雪松醇可抑制DHT诱导的AR核转位;此外,笔者还观察到,DHT可以下调ALP表达并上调AR共激活因子FHL2的表达,而雪松醇可以逆转DHT对成纤维细胞的作用。结论雪松醇除阻止DHT诱导AR核转位以外,还能抑制共激活因子FHL2的表达,双靶点下调DHT-AR信号活性。外用雪松醇治疗雄激素源性脱发(AGA)值得进一步研究。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 CD133 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 30μmol/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 f

关 键 词：雪松醇 雄激素受体 核转位 真皮乳头细胞 雄激素源性脱发 

分 类 号：R758.71[医药卫生—皮肤病学与性病学]