摘要
目的本研究主要观察中药益发复方含药血清对大鼠毛囊干细胞增殖及K19、β1整合素mRNA表达水平的调控作用。方法自制益发复方含药血清；取7～8日龄Wistar大鼠，分离隆突区毛囊，用二步消化酶法获取毛囊外根鞘细胞，再用Ⅳ型胶原黏附法分离毛囊干细胞。将分离得到的毛囊干细胞分为实验组a1-β3（培养液中分别加入10％高、中、低剂量的益发含药血清）、对照组b（培养液中加入10％空白对照血清）及对照组c只加无血清角质细胞培养液（KSFM）。用MTY法制作生长曲线，观察各组培养细胞生长规律和特性；运用K19、β1整合素单克隆抗体免疫组化鉴定培养细胞；RT—PCR检测各组培养细胞K19、β1整合素mRNA表达水平。结果免疫组化鉴定结果显示Ⅳ型胶原黏附细胞符合毛囊干细胞特征。实验组毛囊干细胞克隆形成较早、较多，MTY法检测显示益发血清组毛囊干细胞增生速度明显优于对照组；实验组毛囊干细胞K19、β1整合素基因表达水平较对照组显著提高（P〈0．05），中剂量的益发含药血清（1．65g／mL）作用最为明显。结论中药益发复方能促进毛囊干细胞增生及特异性标记物K19、β1整合素基因表达，这可能是其治疗脱发病及促进毛发生长的药理机制之一。
Objective To investigate the regulatory effects of YiFa compound medicated serum on proliferation of follicle stem cells and mRNA expression of K19 and β1 integrin. Methods Preparated YiFa compound medicated serum using adult Wistar mice; isolated follicle stem cells from 7~8 d Wistar mice by collagen adhesion;group al-a3 adding 10%different concentration YiFa compound medicated serum respectively, group b addingl0% negative medicated serum,group c cultured without medicated serum;identified culture cells by Immunohistoehemistry using K19 and 131 integrin Monoclonal antibody; observed proliferation profile of stem cells by MTI？ growth curve; investigated mRNA expression of K19, 131 integrin by RT-PCR for every groups. Results Immunohistochemistry showed collagen adhesion cells conform to follicle stem cells;YiFa compound serum could obviously promote follicle stem cells proliferation and mRNA expression of K19, β1 integrin compared with controls （P〈O.05）. Conclusion YiFa compound medicated serum could accelerate follicle stem ceils proliferation and K19, β1 integrin mRNA expression,it might be one of the pharmaceutical mechanism for YiFa compound serum in treating alopecia and promoting hair growth.
出处
《中国中西医结合皮肤性病学杂志》
CAS
2013年第6期356-358,共3页
Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine
基金
广东省科技计划课题(项目编号:2010B060900075)
关键词
毛囊干细胞
中药
含药血清
基因表达
Follicle stem cell
Chinese herb
Medicated serum
mRNA expression
