去乙酰化酶抑制剂TSA通过激活ERK和p38抑制间充质干细胞成脂分化  

TSA Inhibits Adipogenic Differentiation in Mesenchymal Stem Cells by Activating ERK and p38 Kinase

作　　者：龚雨琴[1] 季煜华[1] 李智耀[1] 王奎栋[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫实验中心,广州510632

出　　处：《中国生物化学与分子生物学报》2012年第1期35-41,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.30600587;No.30900563;No.81071483);中央高校基本科研业务费专项资金项目;江苏省教育厅自然科学基金项目(No.10KJB180006)~~

摘　　要：本文研究了丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)信号通路在组蛋白去乙酰化酶抑制剂(histone deacetylase inhibitors,HDACis)曲古抑菌素A(trichostatin A,TSA)抑制间充质干细胞(mesenchymal stem cells,MSCs)C3H10T1/2成脂分化中的调节机制.首先利用MTT法检测TSA对其增殖活性的影响;Western印迹法首先检测MAPKs信号通路中pERK和p-p38蛋白在间充质干细胞C3H10T1/2成脂分化过程中的表达情况,以及不同浓度、不同时间TSA处理对pERK和p-p38蛋白差异变化情况;其次再用Western印迹检测TSA对成脂分化过程中间充质干细胞pERK和p-p38蛋白表达的影响.MTT结果显示,TSA浓度在1 nmol/L～100 nmol/L范围内抑制C3H10T1/2细胞的增殖活性,且TSA浓度约为60 nmol/L时即抑制一半以上的C3H10T1/2细胞增殖活性.Western印迹结果显示,TSA处理5 min～80 min,及浓度在1 nmol/L～100 nmol/L范围内激活MAPK信号通路中pERK和p-p38蛋白的表达;C3H10T1/2细胞成脂分化过程中,胞内pERK和p-p38蛋白的表达呈现下调趋势;而TSA抑制了成脂分化过程中C3H10T1/2细胞内pERK和p-p38蛋白的表达变化.本研究结果提示,在C3H10T1/2细胞成脂分化过程中,MAPK信号途径分子pERK和p-p38表达下调;TSA可能是通过活化pERK和p-p38进而抑制间充质干细胞C3H10T1/2成脂分化.This paper was involved in the regulatory mechanism of mitogen activated protein kinases（MAPKs） signaling pathways in the inhibition effect of trichostatin A（TSA） on adipogenic differentiation in mesenchymal stem cells（MSCs）.C3H10T1/2 cells were used as a suitable model for this study.The proliferation of cells treated by TSA was detected by MTT method.Western blotting was first applied to detect the expression of pERK and p-p38 proteins after C3H10T1/2 cells treated with TSA or adipogenic differentiation medium（ADM）,and then to detect the expression of pERK and p-p38 proteins in the adipogenic progress of MSCs after C3H10T1/2 cells treated with TSA.Cells proliferation was inhibited with TSA and inhibition ratio was reached 50% at 60 nmol/L.Western blotting showed that TSA activated pERK and p-p38 proteins in the MAPKs signaling pathways and the expression of pERK and p-p38 proteins were down-regulated in the adipogenic progress.TSA inhibited the expression of pERK and p-p38 proteins in the adipogenic progress were also found in this study.Our results demonstrate that TSA inhibited adipogenic differentiation by activating pERK and p-p38.

关 键 词：曲古抑菌素A(TSA) 丝裂原活化蛋白激酶(MAPKs)信号通路 C3H10T1/2细胞 成脂分化 

分 类 号：R392.12[医药卫生—免疫学]