脂联素对氯化钴诱导肾小管上皮细胞凋亡及炎症反应影响研究

目的 探讨脂联素对氯化钴诱导的肾小管上皮细胞凋亡及炎症反应的影响.方法 将人肾小管上皮细胞株HK-2细胞分入4组:正常对照组,用DME/F-12培养液培养细胞24 h;氯化钴诱导缺氧模型组,用含600.0μM氯化钴的培养液培养细胞24 h;单独脂联素处理组,用含2.5μg/ml脂联素的培养液培养细胞24 h;氯化钴+脂...

Effect of adiponectin on renal tubular epithelial cell apoptosis and inflammation induced by cobalt chloride

Objective To investigate the effect of adiponectin(APN) on the apoptosis and inflammation of renal tubular epithelial cells induced by cobalt chloride(CoCl₂).Methods Human renal tubular epithelial cell line HK-2 cells were divided into 4 groups:normal control group,cultured in DME/F-12 medium for 24 hours;in the CoCl₂ induced hypoxia model group,the cells were cultured in the medium containing 600.0 μM CoCl₂ for 24 hours;in the single APN treatment group,the cells were cultured in the medium containing 2.5 μg/ml APN for 24 hours;in the CoCl₂+APN combined treatment group,the cells were treated with 600.0 μM CoCl₂ and 2.5 μg/ml APN for 24 hours.The morphological changes of HK-2 cells were observed by light microscope,the expression of reactive oxygen species was observed by full spectrum laser confocal microscope,the apoptosis rate was detected by flow cytometry,and the protein expression levels of inflammatory factors such as interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α were detected by Western blot.Results Under light microscope,the cells in normal control group were fusiform with intact cell membrane and good growth.In the hypoxia model group induced by CoCl₂,cell morphology became round,cell shrinkage and cell fragmentation appeared,cell apoptosis and necrosis occurred.There was no change in cell morphology in single APN treatment group.Most of the cells in the CoCl₂+APN treatment group recovered from round to spindle shape,but there were still a few cell fragments.HK-2 cells in normal control group and single APN treatment group showed weak red MitoSox fluorescence.HK-2 cells in the CoCl₂ induced hypoxia model group showed bright red fluorescence,while the intensity of red fluorescence in the CoCl₂+APN combined treatment group was significantly weaker than that in the CoCl₂ induced hypoxia model group.Compared with the normal control group,the apoptosis rate of CoCl₂ induced hypoxia model group and CoCl₂+APN combined treatment group were significantly increased,with statistical significance(P<0.05).Compared with the hypoxia model group induced by CoCl₂,the apoptosis rate of CoCl₂+APN combined treatment group was significantly reduced(P<0.05).Compared with the normal control group,the protein expression levels of IL-1β,IL-6 and TNF-α in the hypoxia model group induced by CoCl₂ increased,P<0.05.Compared with the hypoxia model group induced by CoCl₂,the protein expression levels of IL-1β,IL-6 and TNF-α in the combined treatment group were reduced(P<0.05).Conclusion Adiponectin may inhibit the apoptosis and inflammation of renal tubular epithelial cells induced by cobalt chloride by reducing the production of reactive oxygen species.