Wnt信号通路在压力调控骨髓间充质干细胞膜片成软骨响应中的作用  

Role of Wnt signal pathway in chondrogenic mechanotransduction in bone marrow mesenchymal stem cell sheets

作　　者：黄锦梅[1] 程百祥 马媛媛[1] 刘晓波[1] 张旻 陈永进[1] 

机构地区：[1]第四军医大学口腔医学院,陕西西安710032 [2]西安市第九医院口腔科,陕西西安710054

出　　处：《牙体牙髓牙周病学杂志》2014年第1期1-6,20,共7页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金资助项目(31170888)

摘　　要：目的:研究经典(Wnt/β-catenin,Wnt)信号通路在压力调控骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)膜片成软骨响应中的作用。方法:体外分离、培养兔BMSCs,经鉴定后用含有抗坏血酸的培养基构建BMSCs细胞膜片,并将其随机分2组;对照组在常规条件下培养4 d,实验组施以120 KPa静态压力,1 h/d、连续4 d。然后通过Western Blot检测经典Wnt信号通路相关蛋白的表达水平;Real-time PCR检测Wnt信号通路相关基因和成软骨基因的表达水平。结果:兔BMSCs传代后生长状态稳定,呈梭形;成骨成脂分化诱导实验结果阳性;Western Blot检测显示,实验组β-catenin和p-GSK3β蛋白表达水平均明显高于对照组(P<0.05);Real-time PCR检测显示,实验组经典Wnt信号通路相关基因β-catenin、p-GSK3βmRNA以及成软骨相关基因Col-Ⅱ、Sox-9、Aggrecan mRNA的表达水平均明显高于对照组(P<0.05)。结论:Wnt/β-catenin信号通路介导兔BMSCs膜片在一定的压力刺激下的成软骨响应过程。AIM： To investigate the role of Wnt/β-catenin pathway in the chondrogenic mechanotransdnction in bone marrow mesenchymal stem cell （BMSC） sheets under mechanical pressure. METHODS ： BMSCs were separated, cultured and identified in vitro. BMSC sheets obtained by culture with standard medium containing Vitamin C were randomly divided into blank control group and pressure group. BMSC sheets in pressure group were treated with 120 kPa hydrostatic pressure for lh/day for 4 consecutive days. The Wnt/β-catenin（Wnt） signaling pathway proteins were determined by Western blot. The expression of cartilage related genes was examined by real-time PCR. RESULTS： The subcultured rabbit BMSCs grew well. Osteogenic and adipogenic differentiation ability of the cultured BMSCs were confirmed by induction asays. Western blot showed that the protein expression of 13-catenin and p-GSK3β in pressure group was higher than that in the control group （P 〈 0.05）. Real-time PCR showed that β - catenin, p- GSK3β and chondrogenic markers of Col-II, Sox-9 and aggrecan in pressure-stimulated BMSC sheets were higher than those in the controls （P 〈 0.05）. CONCLUSION： Wnt/β-catenin pathway plays an important role in chondrogenic differentiation of BMSCs under mechanical pressure.

关 键 词：骨髓间充质干细胞膜片 压力 WNT β-catenin 软骨形成 

分 类 号：R780.2[医药卫生—口腔医学]