低氧对类风湿关节炎滑膜成纤维样细胞表达葡萄糖-6-磷酸异构酶及细胞周期的影响  

Hypoxia upregulates glucose-6-phophate isomerase expression and improves proliferation of fibroblast-like synoviocytes from synovium of patients with rheumatoid arthritis

作　　者：陆英[1] 宗明[1] 樊莎莎[1] 卢添宝[1] 戴兴苗 范列英[1] 

机构地区：[1]同济大学附属东方医院检验科,上海200120

出　　处：《中华风湿病学杂志》2015年第5期327-331,I0001,共6页Chinese Journal of Rheumatology

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：目的 研究将通过对骨性关节炎（OA）和RA患者的关节滑膜成纤维样细胞（FLS）进行正常氧和低氧培养后,观察FLS表达葡萄糖-6-磷酸异构酶（G6PI）和缺氧诱导因子（HIF）-1α的变化情况及对细胞周期的影响.方法 原代培养OA和RA患者的滑膜成纤维细胞,分别在正常氧和3％氧环境中培养24 h,实时荧光定量PCR检测G6PI、HIF-1α mRNA表达水平,蛋白印迹法检测G6PI、HIF-1α蛋白表达水平.流式细胞仪检测细胞周期.采用t检验、Mann-Whitney U检验进行统计学处理.结果 与正常氧培养相比,经3％氧培养RA FLS G6PI mRNA的升高倍数（2.6±0.4）明显高于OA FLS的升高倍数（1.5±0.4）（t=4.94,P＜0.05）;RA FLS G6PI蛋白表达水平明显高于OA FLS（t=2.81,P＜0.05）.与正常氧培养相比,RA FLS经3％氧培养HIF-1α mRNA的升高倍数（2.9±0.8）明显高于OA FLS的升高倍数（1.4±0.4）（t=4.51,P＜0.05）;RA FLS HIF-1α蛋白表达水平明显高于OA FLS（t=14.54,P＜0.05）.在3％氧低氧培养环境中,与正常氧培养相比,RA FLS G1期显著减少（t=11.31,P＜0.05）,S期（t=-7.62,P＜0.05）、G2期显著增加（t=-9.95,P＜0.05）.结论 在RA低氧环境中,G6PI、HIF-1αmRNA和蛋白水平在FLS中的表达增高,FLS细胞增殖加快.Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase （G6PI） expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis （RA） and osteoarthritis （OA） under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia （3% oxygen） or normoxia （21% oxygen） for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA （2.6±0.4 vs 1.5±0.4,P〈0.05）.The protein levels of G6PI in RA were higher than those of OA （P〈0.05）.The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA （2.9±0.8vs 1.4 ±0.4,P〈0.05）.The protein levels of HIF-1α in RA were higher than those of OA （P〈0.05）.The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs （t=1 1.31,P〈0.05）.The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.

关 键 词：关节炎 类风湿 葡糖-6-磷酸异构酶 缺氧诱导因子1 Α亚基 细胞周期 成纤维样细胞 

分 类 号：R593.22[医药卫生—内科学]