Thrsp蛋白在青少年特发性脊柱侧弯患者MSCs成脂分化过程中的差异性表达和作用  

Expression of THRSP may be involved in mesenchymal stem cell adipogenic differentiation of adolescent idiopathic scoliosis

作　　者：王其飞[1] 杨军林[1] 范恒伟[1] 黄紫房[1] 林翔[1] 谢超凡[1] 李佛保[1] 

机构地区：[1]中山大学附属第一医院脊柱侧弯中心,广州510080

出　　处：《中国矫形外科杂志》2015年第3期263-269,共7页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81071439)

摘　　要：[目的]比较青少年特发性脊柱侧凸（AIS）患者与正常人骨髓间充质干细胞的成脂分化过程中基因表达的差异。[方法]收集本院脊柱侧弯中心2011年12月～2012年12月收治的AIS患者20例的骨髓血,同时采集10例年龄段性别接近的志愿者骨髓血作为对照组。采用密度梯度离心法提取MSC细胞,培养至P3流式细胞仪定性。诱导成脂分化14 d后提取全RNA进行Affymetrix 3＇IVT表达谱芯片检测。对主要检查结果进行RT-PCR验证,并且对结果中特异性高表达的Thrsp蛋白的表达情况进行Western blot检验。[结果]受试者骨髓血均成功采集分离,密度梯度离心分离MSCs并培养至P3,流式细胞仪验证其性质,成脂分化培养14 d后苏木红染色定性为脂肪细胞,提取全RNA电泳条带清晰,表达谱芯片检测结果显示AIS患者MSC在成脂分化过程中有111条基因表达上调,189条基因表达下调。Western blot检验结果显示Thrsp蛋白在脂肪细胞中高度表达。[结论]AIS来源MSCs成脂分化相关差异表达的300条基因可能参与了AIS的发生和发展,Thrsp基因可能是导致MSCs分化差异性的关键因素。[Objective] To compare gene expression differentiation of the adipogenic process between bone marrow mesenchymal stem cells（ MSCs） of adolescent idiopathic scoliosis（ AIS） patients and healthy patients. [Methods] Twenty patients with AIS treated in our hospital from December 2011 to December 2012 had a bone marrow sample taken along with 10 control group volunteers of similar age and sex. MSCs were extracted by density gradient centrifugation,cultured to P3 cells,and qualitative by flow cytometry. Fourteen days after adipogenic differentiation total RNA was extracted for Affymetrix 3＇ DIVT expression profiling microarrays. The results were confirmed by RT- PCR validation and protein expression of THRSP was validated by the western blot test. [Results] MSCs were successfully collected from the bone marrow and separated by density gradient centrifugation,cultured to P3 cells,and flow cytometry was used to verify the nature of the cells. Adipogenic cells were cultured for 14 days after hematoxylin staining characterized them as fat cells. Extracted total RNA for Affymetrix 3 ＇ IVT microarray expression profiling showed that in patients with AIS MSC adipogenic differentiation of 213 upregulated genes,310 genes were downregulated. Western blot test results show that THRSP proteins are highly expressed in fat cells. [Conclusion] AIS source MSCs adipogenic differentiation of 300 differentially expressed genes may be involved in the occurrence and development of AIS. THRSP may play an important role in the process of differentiation of MSCs.

关 键 词：青少年特发性脊柱侧凸 MSC 成脂分化 Thrsp 

分 类 号：R726.8[医药卫生—儿科]