Ⅰ型胶原及骨膜蛋白在骨质疏松大鼠骨髓基质细胞膜片中的表达研究  

Expression of Collagen-Ⅰ and Periostin in Bone Marrow Stromal Cell Sheet of Osteoporotic Rats

作　　者：李丽生 何梦娇 骆凯[3] 何武兵[1] LI Lisheng;HE Mengjiao;LUO Kai;HE Wubing(First-author’s address:Fujian Provincial Hospital,Fuzhou 350000,China;不详)

机构地区：[1]福建省立医院,福建福州350000 [2]福建省口腔疾病研究重点实验室 [3]福建医科大学附属口腔医院

出　　处：《中外医学研究》2022年第29期151-155,共5页CHINESE AND FOREIGN MEDICAL RESEARCH

基　　金：国家自然科学基金项目(81870766);福建医科大学启航基金项目(2018QH1150)。

摘　　要：目的:探究Ⅰ型胶原(Col-Ⅰ)、骨膜蛋白(POSTN)在骨质疏松大鼠骨髓基质细胞(BMSC)膜片中的表达情况。方法:在体外分离培养骨质疏松大鼠BMSCs,鉴定其克隆形成能力、多向分化能力,采用富含L-抗坏血酸培养基连续诱导构建细胞膜片,诱导5、10 d形成的细胞膜片经HE染色及Masson染色后,观察细胞膜片结构,分别采用qRT-PCR及免疫组织化学染色检测细胞外基质(ECM)中Col-Ⅰ、POSTN在基因及蛋白水平的表达情况。结果:体外分离培养的骨质疏松大鼠BMSCs具有克隆形成能力及成骨、成脂分化能力,L-抗坏血酸连续诱导获取的细胞膜片富含ECM;qRT-PCR结果显示,诱导第10天细胞膜片的ECM相关基因Col-Ⅰ及POSTN的mRNA相对表达量高于第5天(P<0.05);免疫组织化学染色结果显示诱导时间延长,细胞膜片中的ECM相应增多,且高表达Col-Ⅰ及POSTN。结论:PMO大鼠BMSC膜片可保留自身分泌的ECM,细胞膜片富含Col-Ⅰ及POSTN,为后续自体移植细胞膜片实现骨质疏松状态下骨组织再生提供帮助。Objective:To investigate the expression of collagen-Ⅰ (Col-Ⅰ) and periostin (POSTN) in bone marrow stromal cell sheet (BMSC) of osteoporotic rats.Method:BMSCs were isolated and cultured in vitro to identify their cloning formation capability and multipotential differentiation ability,the cell sheets were constructed by continuous induction with L-ascorbic Acid-enriched Medium,cell sheets formed at 5,10 d were stained by HE and Masson to observe the structure,quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical staining were used to detect the mRNA and protein expression of Col-Ⅰ and POSTN.Result:Osteoporotic rat BMSCs were isolated and cultured in vitro which possess cloning formation,osteogenic and adipogenic differentiation ability,the cell sheets obtained by continuous induction of L-ascorbic Acid were rich in ECM.The results of qRT-PCR showed that the mRNA expression of Col-Ⅰ and POSTN were up-regulated on the 10th day compared with the 5th day of the cell sheets (P<0.05).The results of immunohistochemical staining showed that the induction time was prolonged,the ECM in the cell membrane increased correspondingly,and the Col-Ⅰ and POSTN were highly expressed.Conclusion:BMSC sheet retains a large amount of self-secreted ECM and is rich in Col-Ⅰ and POSTN,which can provide assistance for subsequent autologous transplantation of cell sheet to achieve bone tissue regeneration in osteoporosis.

关 键 词：绝经后骨质疏松症 细胞膜片 细胞外基质 胶原 骨膜蛋白 

分 类 号：R580[医药卫生—内分泌]