SAHA抑制间充质干细胞C3H10T1/2的增殖与成脂分化  

SAHA Suppresses the Proliferation and Adipogenic Differentiation of Mesenchymal Stem Cell C3H10T1/2

作　　者：王奎栋[1] 李智耀[1] 吴伟[1] 龚雨琴[1] 季煜华[1] 

机构地区：[1]暨南大学生命科学技术学院、组织移植与实验研究中心,广州510632

出　　处：《中国生物化学与分子生物学报》2012年第6期555-560,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.30600587;.30900563;.81141048;.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)资助~~

摘　　要：本文研究组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)对间充质干细胞(mesenchymal stem cells,MSCs)C3H10T1/2增殖和成脂分化的影响及其可能的作用机制.用Western印迹验证SAHA对细胞内蛋白乙酰化的影响;用MTT和流式细胞术检测细胞活性和细胞周期;利用油红O染色检测细胞成脂分化,实时定量PCR检测PPARγ2和成脂分化标志物Fabp4、perilipin以及adipoq mRNA的转录.Western印迹结果显示,SAHA可促进细胞内蛋白的乙酰化.MTT和流式细胞术结果显示,SAHA对C3H10T1/2细胞活性的抑制呈浓度依赖性,随着SAHA浓度增加,细胞形态趋向于展平,并将细胞周期抑制在G0/G1期;SAHA可呈浓度依赖性抑制C3H10T1/2细胞的成脂分化作用.同时实时定量PCR结果显示,SAHA抑制成脂关键转录因子PPARγ2,脂肪因子Fabp4、perilipin和adipoq mRNA的转录.综上所述,SAHA可影响间充质干细胞C3H10T1/2细胞形态,并呈剂量依赖性地抑制其增殖和成脂分化.To study the effect and underlying mechanism of histone deacetylase （HDAC） inhibitor, suberoylanilide hydroxamic acid （SAHA）, on the proliferation and adipogenic differentiation of C3H10T1/2 mesenchymal stem cell. The effect of SAHA on the acetylation was verified by Western blotting. MTT assay and flow cytometry were used to evaluate the cell viability and cell cycle indexes following treatments with SAHA. The adipogenic differentiation was characterized by oil red O staining. Real-time PCR was used to detect the expression of adipogenic differentiation markers, including PPARγ2, Fabp4, perilipin and adipoq. Western blot showed that SAHA increased the acetylation of intracellular protein. SAHA treatment altered C3H10T1/2 cell morphology, inhibited cell proliferation, and caused G0/G, arrest of the cell cycle in a concentration dependent manner as shown in MTT and FCM measures. Real-time PCR showed the expression of PPARγ2 （key transcription factor of adipogenic differentiation）, Fabp4, perilipin and adipoq were reduced. The results demonstrated that SAHA inhibited the proliferation and adipogenic differentiation of C3H10T1/2 mesenchymal stem cell.

关 键 词：辛二酰苯胺异羟肟酸(SAHA) 间充质干细胞 成脂分化 

分 类 号：R392.12[医药卫生—免疫学]