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  

The study on the mechanism of As2O3 inducing synoviocytes apoptosis of rheumatoid arthritis in vitro

作　　者：韩锋[1] 刘伟丽[1] 任洁[1] 齐文成[1] 

机构地区：[1]天津市第一中心医院风湿免疫科,300192

出　　处：《中华风湿病学杂志》2010年第4期236-239,共4页Chinese Journal of Rheumatology

基　　金：天津市卫生局课题（05ky11）

摘　　要：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（As2O3）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As2O3对体外培养的RA滑膜细胞凋亡的诱导作用，As2O3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。Bcl～2mRNA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As2O3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80μmol／L药物浓度最为明显。凋亡早期（6h）细胞色素C水平升高80μmol／L组升高明显[NC（0．34±0．27），80μmol／L组（32．04±1．62），P〈0．05]；不同浓度的As2O3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对照减少，As2O3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。Objective To investigate the effects of As2O3 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 As2O3. Methods Cultured synoviocytes were incubated with As2O3. The effects of apoptosis were detected by light microscope, electronic microscope and flow cytometry. The level of CtyC was measured by ELISA 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 ENC （0.34±0.27）, RA （32.00±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 （1.08±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 synovioeytes induced by As2O3 than normal control （NC）. Conclusion The apoptosis of synovioeytes could be induced by As2O3, and the level of caspase-3 is increased. The mechanisms of As2O3 induced synoviocytes apoptosis maybe due to the increased level of CtyC and CtyC, and the decreased level of Bcl-2.

关 键 词：关节炎 类风湿 细胞凋亡 滑膜 三氧化二砷 

分 类 号：R593.22[医药卫生—内科学]