缺氧复氧对滑膜细胞IGF-IGFBP-3 以及线粒体的影响

目的:探讨氧波动环境对原代成纤维样滑膜细胞(fibroblast-like synoviocyte, FLS)胰岛素样生长因子-1(insulin growth factor-1,IGF-1)、胰岛素样生长因子结合蛋白-3(insulin-like growth factor binding protein-3, IGFBP-3)及线粒体的影响。方法:分离并鉴定正常人滑膜细胞,再对滑膜细胞进行分组:对照组、缺氧/再充氧(hypoxia/reoxygenation,H/R)干预组。采用实时定量PCR检测滑膜细胞中IGF-1、IGFBP-3的m RNA水平;Western blot检测滑膜细胞中IGF-1、IGFBP-3的蛋白水平;流式细胞仪检测线粒体膜电位(Mitochondrial membrane potential, MMP)以及线粒体通透性转换孔(Mitochondrial Permeability Transition Pore, MPTP)的变化。结果:与对照组比较,H/R干预组的相对IGF-1和IGFBP-3的m RNA水平和蛋白表达水平显著升高(P0.05),膜电位水平降低(P0.05),线粒体通透性转换孔开放。结论:氧波动环境可促进IGF-1和IGFBP-3的表达及细胞线粒体损伤,其可能是骨关节炎(OA)发病的重要机制之一。

Effects of Hypoxia and Reoxygenation on IGF-1, IGFBP-3 and Mitochondria in Synovial Cells

ABSTRACT Objective: To investigate the effects of oxygen-induced fluctuations on the expression of insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein-3 -3, IGFBP-3 and the change of mitochondria. Methods: Synovial cells were isolated and identified, which were divided into control group and Hypoxia/ Reoxygenation (H/R) intervention group. The mRNA levels of IGF-1 and IGFBP-3 in synoviocytes were detected by real-time quantitative PCR; The protein levels of IGF-1 and IGFBP-3 in synoviocytes were detected by Western blot; The mitochondrial membrane potential and mitochondrial permeability transition pore was detected by flow cytometry. Results: Compared with the control group, the mRNA and protein levels of IGF-1 and IGFBP-3 in the H/R intervention group were significantly increased(P<0.05); the membrane potential was significantly lower than that in the control group (P<0.05) and openness of mitochondrial permeability transition pore. Conclusion: The fluctuating environment of oxygen can promote the expression of IGF-1 and IGFBP-3 and the mitochondrial damage, which would be a possible mechanism of osteoarthritis(OA).