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为探索组蛋白去乙酰化酶抑制剂(suberoylanilide hydroxamic acid,SAHA)对经II型胶原体内致敏T细胞的体外增殖影响。以II型胶原致敏C57B6小鼠7 d后,收集引流淋巴结细胞。MTT法观察不同浓度SAHA对CⅡ特异性T细胞增殖的抑制作用;ELISA检测细胞因子表达;Annexin-v观察CII特异性T细胞凋亡格局。Caspase 3试剂盒检测CⅡ特异性T细胞中Caspase 3酶活性。结果显示,CⅡ特异性T细胞的体外增殖被SAHA抑制,呈时间和剂量依赖性。细胞因子格局表现为在SAHA处理后,CⅡ特异性T细胞分泌IL-2、IL-12和IFN-γ明显下降。当SAHA大于500 nmol/L时,CⅡ特异性T细胞凋亡比率增加,进一步的研究表明,当SAHA大于1 000 nmol/L时,CII特异性T细胞中Caspase 3活性明显高于对照组。SAHA能够抑制CII特异性T细胞增殖和细胞因子产生,并可通过上调Caspase 3活性而诱导CⅡ特异性T细胞凋亡,增加其抑制T细胞增殖作用。
To explore the effect of suberoylanilide hydroxamic acid (SAHA) on the proliferation of sensitized T cells via type II collagen in vitro. After 7 days of C57B6 mice sensitized with type II collagen, draining lymph node cells were collected. The inhibitory effect of different concentrations of SAHA on the proliferation of CⅡ-specific T cells was observed by MTT assay. The expression of cytokines was detected by ELISA. The apoptosis of CII-specific T cells was observed by Annexin-v. Caspase 3 kit assay Caspase 3 activity in CII-specific T cells. The results showed that in vitro proliferation of CⅡ-specific T cells was inhibited by SAHA in a time and dose-dependent manner. Cytokine pattern showed that after SAHA treatment, CⅡ-specific T cells secreted IL-2, IL-12 and IFN-γ significantly decreased. When SAHA is greater than 500 nmol / L, the rate of apoptosis of CⅡ-specific T cells is increased. Further studies show that when the SAHA is greater than 1 000 nmol / L, the activity of Caspase 3 in CII-specific T cells is significantly higher than that of the control group. SAHA can inhibit CII-specific T cell proliferation and cytokine production, and can induce CⅡ-specific T cell apoptosis by up-regulating Caspase 3 activity and increase its inhibitory effect on T cell proliferation.