去乙酰化酶抑制剂TSA对间充质干细胞C3H10T1/2增殖和成脂分化的影响  

Effects of histone deacetylase inhibitor trichostatin A on proliferation and adipogenic differentiation of C3H10T1/2 mesenchymal stem cells

作　　者：吴伟[1] 季菊玲[2] 王奎栋[1] 李智耀[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免疫实验中心,广东广州510632 [2]南通大学医学院病理教研室,江苏南通226001

出　　处：《暨南大学学报（自然科学与医学版）》2011年第3期328-333,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30600587;3090056;381071483);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(10KJB180006);南通市科技局项目(K2009028);南通大学自然科学项目(07Z089)

摘　　要：采用MTT和流式细胞术分别检测不同浓度的TSA对C3H10T1/2细胞活性和细胞周期分布的影响;油红O染色检测TSA对其成脂分化的影响,实时定量PCR检测TSA对成脂分化的关键转录因子PPAR-γ,以及成脂分化标志物Fabp4和Adipoq mRNA转录的影响.研究去乙酰化酶抑制剂TSA对间充质干细胞C3H10T1/2增殖和成脂分化的影响及其可能的作用机制.结果显示TSA浓度为1、10和30 nmol/L呈浓度依赖性地抑制C3H10T1/2细胞活性,改变细胞形态,并将其细胞周期抑制在G0/G1期;TSA浓度为10nmol/L明显抑制C3H10T1/2细胞的成脂分化作用,并呈浓度依赖性地抑制PPAR-γ、Fabp4和Adipoq mRNA的转录.表明TSA呈剂量依赖性地抑制间充质干细胞C3H10T1/2的增殖和成脂分化,除转录水平调控外,非组蛋白如细胞骨架相关蛋白可能也参与TSA的抑制作用.To investigate the effects of histone deaetylase（HDA） inhibitor trichostatin A on proliferation and adipogenie differentiation of C3H10T1/2 mesenehymal stem cells as well as the underlying mechanism. MTT assay and flow cytometry-based cell cycle analysis were used to evaluate the effects of triehostatin （TSA） at different concentrations on cell viability and cell cycle of cultured C3H10T1/2 cells. The influenee of serial concentrations of TSA on adipogenie differentiation of C3H10T1/2 cells was determined by oil red O staining. The effects of TSA on the expression of PPAR-γ, a key adipogenic transcription faetor, and the Fabp4 and Adipoq, the two adipogenie differentiation markers, were determined by realtime PCR. Our results showed that the TSA inhibited the proliferation of C3H10T1/2 cells in a dose-de- pendent manner and the cell cycle of C3H10T1/2 cells was arrested at G0/G1 phase. Moreover, the morphology of C3H10T1/2 cells was found to be flattened by treatment with TSA. The adipogenie differentiation of C3H10T1/2 cells and the expression of PPAR-γ, Fabp4 and Adipoq were suppressed by TSA in a dose-dependent manner. Our study suggested that TSA inhibit the proliferation and adipogenic differentiation of C3H10T1/2 mesenchymal stem cells; in addition to transcriptional regulation, non-histone proteins such as cytoskeleton related proteins, might also be involved in this process.

关 键 词：曲古抑菌素A 间充质干细胞 成脂分化 增殖 

分 类 号：R392.12[医药卫生—免疫学]