摘要
【目的】探究不同浓度双氢睾酮(DHT)对毛囊生长和毛囊细胞增殖的影响及其与miR-133b表达的关系,然后进一步探究miR-133b对毛乳头细胞的增殖和诱导能力的影响。【方法】体外显微分离人毛囊,选取生长期的毛囊进行培养,显微镜下每日测量并拍摄不同浓度的DHT处理组(10-8、10-7、10-6和10-5 mol/L)和对照组各组毛囊的生长长度,免疫荧光检测各组毛母质细胞Ki-67表达量评估毛囊增殖能力,qRT-PCR检测各组毛囊中miR-133b的表达量。通过Lipofectamine 2000转染miR-133b mimics及miR-133b NC至人毛乳头细胞中,CCK-8检测各组人毛乳头细胞增殖能力,qRT-PCR及Western Blot检测毛乳头细胞诱导能力指标Versican、ALP、β-catenin的mRNA及蛋白水平的相对表达量。【结果】与对照组相比,DHT 10-5 mol/L实验组对毛囊的生长的抑制作用有统计学意义(P<0.05),DHT 10-5 mol/L实验组进入退行期的时间提前;其他组生长长度与对照组相比无统计学意义。免疫荧光及qRT-PCR显示,与对照组相比,DHT 10-5 mol/L实验组毛母质细胞中Ki-67阳性细胞较少且毛囊中miR-133b的相对表达量明显升高,相当于对照组的(3.17±0.26)倍(P<0.01)。CCK-8显示,miR-133b mimics组OD450低于miR-133b NC组(P<0.05);qRT-PCR显示,在mRNA水平,miR-133b mimics组Versican、ALP、β-catenin的相对表达量均低于miR-133b NC组(P<0.001、P<0.01和P<0.01)。Western blot显示,在蛋白水平,miR-133b mimics组Versican、ALP、β-catenin的相对表达量同样均低于miR-133b NC组(P<0.01、P<0.001和P<0.01)。【结论】高浓度DHT可能通过调控miR-133b的表达抑制人毛乳头细胞的增殖活性及诱导能力,最终抑制人毛囊的生长及增殖。
【Objective】To explore the effects of different concentrations of 5α-dihydrotestosterone(DHT)on growth and proliferation of human hair follicles(HFs)and their relationship with miR-133b expression,then further explore the role of miR-133b in the proliferation and inducibility of human dermal papille cells【.Methods】HFs were isolated by micro⁃dissection,then the anagen isolated HFs were cultured and divided into different concentrations of DHT treatment groups(10-8mol/L,10-7mol/L,10-6mol/L,10-5mol/L)and the blank control group.The HF growth and morphology were measured and evaluated.The expression of Ki-67 in hair matrix cells and miR-133b were detected by immunofluorescence assay and qRT-PCR respectively.Lipofectamine 2000 was used to transfect miR-133b mimics and miR-133b NC into human dermal papilla cells.CCK-8 was used to assess the proliferation ability of human dermal papilla cells.qRT-PCR and West⁃ern Blot were performed to evaluate respectively the mRNA and protein levels of the markers associated with inductive abil⁃ity of dermal papilla cells,such as Versican,ALP andβ-catenin.【Results】Compared with the control group,no other treatment groups but the DHT 10-5mol/L group showed statistically significant inhibitory effect on HF growth(P<0.05).The catagen in the DHT 10-5mol/L group appeared earlier than that in the control group and there was no statistically signif⁃icant difference in HF growth between other treatment groups and the control group.The DHT 10-5mol/L group showed low⁃er percentage of Ki-67-positive cells in hair matrix cells and significantly increased relative expression of miR-133b in HFs,3.17±0.26 times more than that in the control group(P<0.01).CCK-8 assay revealed that the OD450 value in the miR-133b mimics group was lower than that in the miR-133b NC group(P<0.05).qRT-PCR revealed that the mRNA ex⁃pression levels of Versican,ALP,and β-catenin in the miR-133b mimics group were all lower than those in the miR-133b NC group(P<0.001,P<0.01,P<0.01).Western Blot reve
作者
邓文佳
邓长健
韩乐
刘本
唐鑫
万苗坚
DENG Wen-jia;DENG Chang-jian;HAN Le;LIU Ben;TANG Xin;WAN Miao-jian(Department of Dermatology and Venereology,The Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China;Department of Otorhinolaryngology,the People's Hospital of Longgang District,Shenzhen 518172,China)
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2021年第2期202-208,共7页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(81872542,81271769)。
