CAJ | 학술논문

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.
목적 연구장통과대골성관절염(OA)화RA환자적관절활막성섬유양세포(FLS)진행정상양화저양배양후,관찰FLS표체포도당-6-린산이구매(G6PI)화결양유도인자(HIF)-1α적변화정황급대세포주기적영향.방법 원대배양OA화RA환자적활막성섬유세포,분별재정상양화3％양배경중배양24 h,실시형광정량PCR검측G6PI、HIF-1α mRNA표체수평,단백인적법검측G6PI、HIF-1α단백표체수평.류식세포의검측세포주기.채용t검험、Mann-Whitney U검험진행통계학처리.결과 여정상양배양상비,경3％양배양RA FLS G6PI mRNA적승고배수(2.6±0.4)명현고우OA FLS적승고배수(1.5±0.4)(t=4.94,P＜0.05);RA FLS G6PI단백표체수평명현고우OA FLS(t=2.81,P＜0.05).여정상양배양상비,RA FLS경3％양배양HIF-1α mRNA적승고배수(2.9±0.8)명현고우OA FLS적승고배수(1.4±0.4)(t=4.51,P＜0.05);RA FLS HIF-1α단백표체수평명현고우OA FLS(t=14.54,P＜0.05).재3％양저양배양배경중,여정상양배양상비,RA FLS G1기현저감소(t=11.31,P＜0.05),S기(t=-7.62,P＜0.05)、G2기현저증가(t=-9.95,P＜0.05).결론 재RA저양배경중,G6PI、HIF-1αmRNA화단백수평재FLS중적표체증고,FLS세포증식가쾌.
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.