己酮可可碱联合骨髓间充质干细胞对高糖诱导的肾小球系膜细胞炎症因子及NF-κB信号通路的影响

Effect of Pentoxifylline Combined with Bone Marrow Mesenchymal Stem Cells on Inflammatory Factors and NF-κB Signaling Pathways in Glomerular Mesangial Cells Induced by High Glucose

关键词:  己酮可可碱  骨髓间充质干细胞  肾小球系膜细胞  炎症因子  NF-ĸB信号通路

Key words:  pentoxifylline  bone marrow mesenchymal stem cells  glomerular mesangial cells  inflammatory factors  NF-κB signaling pathway

摘要:

目的 探讨己酮可可碱（pentoxifylline，PTX）联合骨髓间充质干细胞（mesenchymal stem cells，MSCs）对高糖作用下肾小球系膜细胞炎症因子及NF-κB信号通路的影响。方法 体外高糖培养小鼠肾小球系膜细胞，分为6组：对照组、糖尿病肾病（diabetic nephropathy，DN）组、MSCs组、MSCs+PTX 0.1 mmol·L^-1组、MSCs+PTX 0.3 mmol·L^-1组和MSCs+PTX 1 mmol·L^-1组。采用ELISA检测各组系膜细胞IL-6和TNF-α含量，采用Western blotting检测各组系膜细胞中NF-κB p65，IKKα及IKKβ蛋白的表达。结果 与对照组相比，DN组系膜细胞的IL-6和TNF-α含量显著增高（P<0.05），NF-κB p65、IKKα及IKKβ蛋白相对表达量显著升高（P<0.05）。与DN组相比，MSCs组及MSCs组联合不同浓度PTX干预组的系膜细胞分泌的IL-6及TNF-α含量出现明显降低（P<0.05），NF-κB p65、IKKα及IKKβ蛋白表达量显著降低（P<0.05）。与MSCs组比较，MSCs+PTX 0.1 mmol·L^-1组、MSCs+PTX 0.3 mmol·L^-1组及MSCs+PTX 1 mmol·L^-1组的IL-6和TNF-α含量显著下降（P<0.05），NF-κB p65、IKKα和IKKβ的蛋白相对表达量明显降低（P<0.05），且呈现PTX浓度依赖性。结论 PTX联合MSCs通过抑制高糖诱导下肾小球系膜细胞中NF-κB信号通路的表达及炎症因子水平对DN具有一定的治疗作用。

Abstract:

OBJECTIVE To explore the effect on pentoxifylline(PTX) combined with bone marrow mesenchymal stem cells (MSCs) on inflammatory factors and NF-κB signaling pathways of glomerular mesangial cells induced by high glucose. METHODS Mice mesangial cells were cultured in high glucose in vitro and divided into 6 groups:control group, DN(diabetic nephropathy) group, MSCs group, MSCs+PTX 0.1 mmol·L^-1 group, MSCs+PTX 0.3 mmol·L^-1 group and MSCs+PTX 1 mmol·L^-1 group. The levels of IL-6 and TNF-α in mesangial cells of each group were detected by ELISA. The expressions of NF-κB p65, IKKα and IKKβ proteins in mesangial cells were detected by Western blotting. RESULTS Compared with the control group, the levels of IL-6 and TNF-α in the injured mesangial cells of DN group were significantly increased(P<0.05), and the relative expression of NF-κB p65, IKKα and IKKβ proteins were significantly increased(P<0.05). Compared with the DN group, the levels of IL-6 and TNF-α secreted by mesangial cells in MSCs group and MSCs combined with different concentrations of PTX groups were significantly decreased(P<0.05), and the expression levels of NF-κB p65, IKKα and IKKβ proteins were significantly decreased (P<0.05). Compared with the MSCs group, IL-6 and TNF-α levels in MSCs+PTX 0.1 mmol·L^-1 group, MSCs+PTX 0.3 mmol·L^-1 group and MSCs+PTX 1 mmol·L^-1 group were significantly decreased(P<0.05), and the relative expression levels of NF-κB p65, IKKα and IKKβ were significantly decreased(P<0.05), and it was PTX concentration dependence. CONCLUSION PTX combined with MSCs can treat DN by inhibiting the expression of NF-κB signaling pathway and the level of inflammatory factors in glomerular mesangial cells induced by high glucose.