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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.
This article studies the effects of suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on the proliferation and adipogenic differentiation of mesenchymal stem cells (MSCs) C3H10T1 / 2 and its possible mechanism The effect mechanism of SAHA on protein acetylation was verified by Western blotting. The cell viability and cell cycle were detected by MTT and flow cytometry. The adipogenic differentiation was detected by oil red O staining. Real-time quantitative PCR was used to detect the expression of PPARγ2 and Lipid differentiation markers Fabp4, perilipin and adipoq mRNA transcription.Western blotting results show that SAHA can promote intracellular protein acetylation.MTT and flow cytometry results showed that SAHA C3H10T1 / 2 cell activity in a concentration-dependent manner , With the increase of SAHA concentration, cell morphology tended to flatten and the cell cycle was inhibited in G0 / G1 phase. SAHA could inhibit the adipogenic differentiation of C3H10T1 / 2 cells in a concentration-dependent manner.At the same time, real-time quantitative PCR results showed that SAHA Inhibited the transcription of PPARγ2, Fabp4, perilipin and adipoq mRNA, which are the key transcriptional factors of adipogenic differentiation.Conclusion SAHA can affect the morphology of mesenchymal stem cells C3H10T1 / 2 Dose-dependently inhibited adipogenic differentiation and proliferation.