缺氧复氧对滑膜细胞IGF-IGFBP-3以及线粒体的影响

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

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(P<0.05),膜电位水平降低(P<0.05),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。

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 m RNA 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 m RNA 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）.