三氧化二砷对类风湿关节炎滑膜细胞凋亡影响的体外研究

目的 探讨类风湿关节炎(RA)患者滑膜细胞的凋亡异常,以及三氧化二砷(As_2O_3)对体外培养RA的滑膜细胞凋亡的诱导作用.方法 用光镜、电镜、流式细胞仪检测As_2O_3对体外培养的RA滑膜细胞凋亡的诱导作用,As_2O_3,作用于RA滑膜后酶联免疫吸附试验(ELISA)检测凋亡过程中细胞色素C的变化,反转录-聚合酶链反应(RT-PCR)检测Caspase-3.Bel-2 mRNA表达,采用单因素方差分析和LSD-t检验、Dunnet-t检验进行统计学处理.结果 流式细胞仪检测发现不同浓度的As_2O_3作用后,滑膜细胞的凋亡较空白对照组(NC)增加NC(0.95±0.87)%,RA(0.21±0.12)%,P<0.05],呈一定的剂量依赖性,尤其以80 μmol/L药物浓度最为明显.凋亡早期(6 h)细胞色素C水平升高80 μmol/L组升高明显NC(0.34±0.27),80 μmol/L组(32.04±1.62),P<0.05];不同浓度的As_2O_3作用后细胞中Caspase-3 mRNA表达显著增强NC(0.144±0.022),RA(0.323±0.047),(0.824±0.109),(1.213±0.196),P<0.05],Bcl-2的mRNA表达显著减弱NC(1.08±0.23),RA(0.94±0.15),(0.46±0.08),(0.22±0.06),P<0.05].结论 RA患者滑膜细胞凋亡较健康埘照减少,As_2O_3通过升高细胞色素c及Caspase-3,抑制Bcl-2诱导RA滑膜细胞凋亡.

The study on the mechanism of As_2O_3 inducing synoviocytes apoptosis of rheumatoid arthritis in vitro

Objective To investigate the effects of As_2O_3 on the apoptosis of synoviocytes in rheumatoid arthritis in vitro and to examine the levels of CtyC, and mRNAs of Caspase-3, Bcl-2 in RA synoviocytes induced by As_2O_3. Methods Cultured synoviocytes were incubated with As_2O_3. The effects of apoptosis were detected by light microscope, electronic microscope and flow cytometry. The level of CtyC was measured by ELJSA and the level of Caspase-3, Bcl-2 mRNAs were detected by RT-PCR. the data were analyzed with ony-way ANOVA, LSD-t test and Dunnet-t test. Results The ratio of the apoptosis cells was increased NC (0.95±0.87)%, RA (0.21 ±0.12)%, P<0.05], the levels of CtyC NC (0.34±0.27), RA (32.04±1.62), P< 0.05] and Caspase-3 mRNAs NC (0.144±0.022), RA (0.323±0.047), (0.824±0.109), (1.213±0.196), P< 0.05] were increased and the level of Bcl-2 mRNA (NC (l.08±0.23), RA (0.94±0.15), (0.46±0.08), (0.22±0.06), P<0.05 )was decreased during the apoptosis of RA synoviocytes induced by As_2O_3 than normal control (NC). Conclusion The apoptosis of synoviocytes could be induced by As_2O_3, and the level of caspase-3 is increased. The mechanisms of As_2O_3 induced synoviocytes apoptosis maybe due to the increased level of CtyC and CtyC, and the decreased level of Bcl-2.