G6PI对阿霉素诱导类风湿关节炎关节滑膜细胞凋亡的保护作用  

G6PI protects synoviocytes from adriamycin-induced apoptosis in rheumatoid arthritis

作　　者：宗明[1,2] 张慧[1,2] 虞珊珊[1,2] 黄斌[1,2] 傅之妍 戴兴苗 范列英[1,2] 

机构地区：[1]同济大学附属东方医院 [2]上海市东方医院检验科,上海200120

出　　处：《现代免疫学》2014年第3期220-225,共6页Current Immunology

基　　金：国家自然科学基金(81373203);上海市科委生物医药领域重点项目(11DZ1973802);上海市优秀学术带头人计划(12XD1404300)

摘　　要：研究葡萄糖-6-磷酸异构酶(glucose-6-phosphate isomerase,G6PI)对阿霉素(adriamycin,ADR)诱导类风湿性关节炎(rheumatoid arthritis RA)滑膜成纤维样细胞(fibroblast-like synoviocytes,FLS)凋亡的影响,探讨其作用机制。本研究从RA的关节滑膜组织中分离培养FLS,利用ADR建立细胞凋亡模型,通过流式细胞仪检测G6PI对细胞凋亡的影响,并通过western blot、Q-PCR检测细胞凋亡相关蛋白Bax、Bcl2、Caspase 3、Fas、Cyclin B1的表达;收集细胞培养上清,通过ELISA方法测定IL-1β和TNF-α含量。结果显示,G6PI(1μg/ml)和G6PI(10μg/ml)组RA-FLS的凋亡指数均显著低于ADR对照组;G6PI(1μg/ml)组与G6PI(10μg/ml)组间无显著差异。ADR凋亡模型中,Caspase 3、Fas和Bax表达明显上调,Cyclin B1下调;G6PI作用后,Caspase 3和Fas表达明显低于对照组,Cyclin B1表达则明显高于对照组。G6PI处理组的细胞培养上清中IL-1β和TNF-α含量均显著高于ADR对照组。研究表明G6PI可能通过Fas介导的Caspase通路来保护FLS免遭凋亡,同时促进RA-FLS分泌炎症因子IL-1β和TNF-α,从而参与RA的发生与发展。To study the influence of glucose-6-phosphate isomerase （G6PI） on adriamycin （ADR） induced apoptosis of fibro- blast-like synoviocytes （FI.S） in patients with rheumatoid arthritis （RA）, and investigate the mechanism involved, the FLS were isolated from the joint synovial tissue of RA patients and cultured. The influence of G6PI on ADR-indueed apoptosis was detected by flow cytometry. The expressions of apoptosis-related protein, Bax, Bcl2, Caspase3, FasL, Fas, survivin and Cyc- lin B1 were determined by Western blot and Q-PCR. The cell culture supernatant was collected and detected for IL-1β and TNF-αby ELISA. The results showed that the apoptosis indices for RA-FLS in G6PI（1/μg/ml） group and G6PI （10/μg/ml） groups were significantly lower than that in ADR group, and this inhibition was not independent on the dosage of ADR. In the ADR apoptosis model, Caspase 3, Fas and Bax proteins were significantly up-regulated, and Cyclin B1 protein was down-regu- lated. In the G6PI group, Caspase3 and Fas proteins were up-regulated, but were lower than that in ADR group, and Cyclin B1 protein was up-regulated. IL-1β and TNF-α in the culture supernatant of RA-FLS were significantly higher than those in ADR group. It is concluded that G6PI might be modulate cell apoptosis by Caspase pathway mediated by Fas. G6PI can induce the secretion of inflammatory cytokines IL-1β and TNF-α. G6PI might play a pathological role in the development of RA.

关 键 词：类风湿性关节炎 成纤维样细胞 葡萄糖一6一磷酸异构酶 细胞凋亡 

分 类 号：R392[医药卫生—免疫学]