大鼠脂肪源干细胞中骨形态发生蛋白信号通路相关分子的表达  

Expression of bone morphogenetic protein signaling pathway related molecules in rat adipose-derived stem cells

作　　者：余美春[1] 杨会营[1] 陶晖[1] 杨春[1] 曲戎梅[1] 原林[1] 余磊[1] 戴景兴[1] 

机构地区：[1]南方医科大学人体解剖学教研室,广东省组织构建与检测重点实验室,广州510515

出　　处：《解剖学杂志》2011年第2期168-171,194,共5页Chinese Journal of Anatomy

基　　金：国家重点基础研究计划（973计划）项目（2007CB512705）;国家自然科学基金青年面上项目（30801464）

摘　　要：目的：探讨大鼠脂肪源干细胞（ADSCs）成骨和成脂肪分化能力及骨形态发生蛋白（BMP）信号通路相关分子的表达.方法：切取Wistar雌性大鼠腹股沟脂肪垫,用酶消化法培养ADSCs.将第4代ADSCs经成骨和成脂肪诱导分化培养后,进行相应的茜素红和油红O染色,以鉴定ADSCs的分化潜能.同时取培养的第4代ADSCs进行流式细胞术检测部分表面抗原.提取ADSCs的总RNA进行RT-PCR,检测BMP信号通路相关分子mRNA的表达.结果：光学显微镜下观察贴壁ADSCs形态多呈梭形、多边形,细胞大小均匀;流式细胞术鉴定ADSCs的表面抗原显示CD90、CD106呈阳性,CD45、CD49d呈阴性;成脂诱导培养14d后,油红O染色显示特异性脂滴,而阴性对照孔中未观察到红染的脂滴.成骨诱导培养21d后,茜素红染色显示矿化结节呈特异性的粉红色,阴性对照孔中未见被染成粉红色的钙结节;RT-PCR检测结果显示BMP信号通路相关分子Bmpr1a、Bmpr1b、Bmpr2; Smad1、Smad5、Smad8的mRNA均阳性表达.结论：ADSCs具有多向分化潜能,可向成骨和成脂肪分化;ADSCs可能存在BMP信号通路.Objective： To investigate the osteogenic and adipogenic potentiality and the expression of bone morphogenetic protein （BMP） signaling pathway related molecules in adipose-derived stem cells （ADSCs）. Methods： ADSCs were obstained from the rats inguinal fat pad and were cultured by collagenase digestion. Passage 4th ADds were cultured in osteogenic medium and adipogenic medium for osteoblast-inducing differentiation and adipogenic differentiation. Osteoblast phenotype and adipogenic differentiation was correspondingly identified with alizarin red staining and oil red O staining. ADds surface antigens were identified by flow cytometry. The total RNA were extracted from ADSCs for detecting the expression of BMP signaling pathway related molecules mRNA by RT-PCR. Results： Under the optical microscope, ADSCs presented morphological fusiform shape, or polygon, and cells size was of uniformity; Surface antigens of ADSCs were positive in CD90 and CD106, negative in CD45 and CD49d; After 14-day culture in adipogenic medium, ADSCs showed lipid droplets by oil red O staining, while stained lipid droplets were not observed in the negative control. ADSCs cultured in osteogenic medium for 21 days. Alizarin red staining was used to show mineralized nodule, while no calcium was observed in the negative control hole; RT-qPCR analysis showed that the expressions of Bmprla, Bmprlb, Bmpr2, Smadl, Smad5 and Smad8 mRNA were positive. Conclusion： ADSCs with pluripotent can be induced into osteogenic and adipogenic differentiatiom ADSCs may present BMP signaling pathway.

关 键 词：脂肪源干细胞 骨形态发生蛋白受体 SMADS 信号通路 诱导分化 筋膜学 

分 类 号：R68[医药卫生—骨科学]