激活PPARγ途径抑制骨髓基质干细胞向成心肌.doc

PAGE PAGE 4 G0S2基因参与调节鼠骨髓基质细胞向脂肪细胞分化 林先和1，彭佑华2，李隆贵3 （1：安徽医科大学第一附属医院心内科，合肥市 230022 2：解放军306医院心内科，北京市 100 3：第三军医大学新桥医院心内科，重庆市 400038） 摘　要：目的　探讨G0S2在调节鼠骨髓基质细胞向脂肪细胞分化过程中的作用。方法 利用原代培养小鼠MSC，脂质体转染法将表达质粒pCAG-PPARγ2及含报告基因的pGL2－COL1A质粒共转染至MSC，G418筛选，RT－PCR检测PPARγ及G0S2 mRNA表达，免疫组化检测PPARγ的表达，westen blot检测G0S2的表达，油红O染色检测细胞内脂滴。结果 MSC在未分化状态下不表达G0S2，经PPARγ基因转染后，G0S2开始表达，转染细胞最终分化为脂肪细胞。结论 PPARγ通过调节G0S2启动子调节后者表达，进而调节MSC向脂肪细胞分化，G0S2基因参与调节MSC向脂肪细胞分化。 关键词：骨髓基质细胞；PPARγ；G0S2 The effects of G0S2 on the differentiation of mouse mallow stroma cells into adipocytes Lin Xian-he1,PENG You-hua2, LI Long-gui3 (1:cardiology devision, the First Affiliated Hospital, Anhui Medical University, Hefei 230022, 2:cardiology devision, the 306 Hospital, PLA Beijing100101, 3:cardiology devision, Xinqiao Hospital, Third military Medical University, Chongqing 400038, China) Abstract: Objective To explore the effects of G0S2 in the differentiation of mouse mallow stroma cells (MSCs) into adipocytes. Methods The primary MSCs were cultured. The eukaryotic expression plasmid vector pCAG-PPARγ2 and pGL2－COL1A reporter vector were cotransfected into MSCs with lipotransfection method followed by G418 selection. The expression levels of PPARγmRNA and G0S2 mRNA were detected by RT-PCR. Immunohistochemistry and Westen blot were emplored to study the protein expression of PPARγ and G0S2. Lipid droplets in the cells were stained with Oil Red O. Results No expression of G0S2 mRNA was found in undifferentiated MSCs. The G0S2 mRNA expressed in the MSCs after transfected with pCAG-PPARγ2, and the cells finaly differentiated into adipocytes. Conclusion PPARγ stimulated G0S2 promoter activity via the PPRE in vivo.Then the expressions of G0S2 can regulate the adipocyte differentiaiton of MSCs. Key words：mallow stroma cells；PPARγ；G0S2 来源于中胚层的骨髓基质细胞(MSC，marrow strom cell)是多能干细胞，具有较强的增殖和分化能力[1]。MSC在分化过程中受诸多核转录因子调控，且调控的基因不同，分化方向不同。如何使植入细胞尽可能向预期的功能细胞分化，是细胞治疗领域的关键性问题之一。因此，迄今为止，其具体的调控机制仍不清楚。因此，深入了解MSC增殖及分化的调控机制有重要意义。 G0S2 (G0/G1 s