疗脱方-毛乳头细胞源性外泌体体外干预雄激素性脱发机制  

Mechanism of Liaotuo Formula-dermal papilla cell-exosomes in vitro intervention of androgenetic alopecia

作　　者：沈琰 刘晓谷[1] 侯欣 郭祥 陈家昊 黄仁杰 王烨航 余王琴 宋红[1] SHEN Yan;LIU Xiaogu;HOU Xin;GUO Xiang;CHEN Jiahao;HUANG Renjie;WANG Yehang;YU Wangqin;SONG Hong(Basic Medical School,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学基础医学院,杭州310053

出　　处：《中华中医药杂志》2024年第12期6769-6775,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：浙江省基础公益研究计划(No.GF20H270032)。

摘　　要：目的:探究疗脱方含药血清作用毛乳头细胞(DPC)源性外泌体(EXO)调控雄激素性脱发(AGA)细胞模型的作用机制。方法:从C57BL/6小鼠须垫提取DPC,CCK8法获取疗脱方含药血清的最佳浓度后处理3代DPC,采用低温超速离心法分离外泌体。以双氢睾酮(DHT)处理DPC法建立AGA体外细胞模型,分别用疗脱方含药血清、非那雄胺含药血清、毛乳头细胞源性外泌体(DPC-EXO)和疗脱方-DPC-EXO对其进行干预。ELISA检测模型中雄激素受体(AR)水平,镜下观察各组细胞生长形态,CCK8法检测各组DPC增殖率,RT-qPCR、Western Blot分别检测骨形态发生蛋白4(BMP4)、音猬因子(SHH)、糖原合酶激酶-3β(GSK-3β)mRNA、蛋白表达情况。结果:模型组DPC呈漩涡状凝集性生长,且AR水平较空白组显著升高(P<0.01);CCK8结果显示,各治疗组的增殖率较模型组显著升高(P<0.01);Western Blot显示,与模型组比较,非那雄胺组、疗脱方组、DPC-EXO组、疗脱方-DPC-EXO组中BMP4、GSK-3β蛋白表达显著降低(P<0.01),非那雄胺组、疗脱方组、疗脱方-DPC-EXO组中SHH蛋白表达显著升高(P<0.01);RT-qPCR显示,各治疗组DPC中BMP4和GSK-3βmRNA较模型组表达显著下调(P<0.01,P<0.05),SHH mRNA表达显著上调(P<0.05,P<0.01)。结论:疗脱方-DPC-EXO对AGA的干预作用可能通过调控BMP4和SHH表达来抑制凋亡蛋白GSK-3β表达,从而促进DPC增殖,抑制DPC凋亡,帮助恢复毛发生长。Objective:To investigate the mechanism of action of serum containing Liaotuo Formula on the regulation of androgenic alopecia(AGA)cell model by dermal papilla cell(DPC)-derived exosomes.Methods:The DPC were extracted from the whisker pads of C57BL/6 mice,and three generations of DPC were treated with the CCK8 method to obtain the optimal concentration of drug-containing serum of Liaotuo Formula,and the exosomes were isolated by low-temperature ultracentrifugation method.An in vitro model of AGA was established by treating DPC with DHT and intervening with Liaotuo Formula-containing serum,finasteride-containing serum,DPC-derived exosomes(DPC-EXO),and Liaotuo Formula-DPC-EXO,respectively.ELISA was used to detect the level of androgen receptor(AR)in the model,cell growth morphology was observed under microscope in each group,CCK8 method was used to detect the proliferation rate of DPC in each group,and RT-qPCR,Western Blot were used to detect the mRNA and protein levels of BMP4,SHH and GSK-3β,respectively.Results:DPC in the model group showed whirling agglutinative growth and significantly higher AR level than the blank group(P<0.01),CCK8 results showed that the proliferation rate of each treatment group was higher than that of the model group(P<0.01),Western Blot showed that the expression levels of BMP4 and GSK-3βprotein in the finasteride group,the treatment group,the DPC-EXO group and the treatment group-DPC-EXO group significantly decreased compared with the model group(P<0.01),the expression of SHH protein in the finasteride group,the treatment group and the treatment group-DPC-EXO group significantly increased(P<0.01);RT-qPCR showed that in the DPC of each treatment group,the expression of BMP4 and GSK-3βmRNA was significantly downregulated(P<0.01,P<0.05),and the expression of SHH mRNA was significantly upregulated than that in model group(P<0.05,P<0.01).Conclusion:The intervention effect of Liaotuo Formula-DPC-EXO on AGA may involve the regulation of BMP4 and SHH expression,leading to the inhibition of ap

关 键 词：外泌体 毛乳头细胞 雄激素性脱发 骨形态发生蛋白4 音猬因子 糖原合酶激酶-3Β 疗脱方 机制 

分 类 号：R73[医药卫生—肿瘤]