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目的观察二甲双胍对系膜细胞(MCs)核因子-κB(NF-κB),单核细胞趋化蛋白-1(MCP-1)表达合成的影响,并探讨其肾脏保护机制。方法体外培养MCs,随机分为正常(Con)组、高糖(HG)组及高糖+不同浓度二甲双胍(M1、M2、M3)组。48h后收集各组细胞及上清液,采用荧光实时定量RTPCR检测细胞NF-κB,MCP-1mRNA含量,Western blot检测NF-κBp65蛋白水平,ELISA检测上清MCP-1蛋白浓度。结果 (1)与Con组比较,HG组NF-κB mRNA和MCP-1mRNA的表达增加(P<0.05);NF-κBp65的相对表达量增加[(1.04±0.01)vs(0.51±0.15),P<0.05],上清MCP-1水平增高[(561.99±23.24)vs(480.73±35.40)pg/ml,P<0.05];(2)二甲双胍干预后,NF-κB mRNA、MCP-1mRNA表达及NF-κBp65蛋白、上清MCP-1含量均下降(P<0.05)。结论二甲双胍可抑制MCs NF-κB和MCP-1的表达合成,可能与其肾脏保护作用有关。
Objective To observe the effect of metformin on the expression of nuclear factor-κB (NF-κB) and monocyte chemoattractant protein-1 (MCP-1) in mesangial cells (MCs) Methods MCs were cultured in vitro and randomly divided into normal group (Con group), high glucose (HG) group and high glucose + metformin group (M1, M2, M3). The cells and supernatants were collected 48h later. The expression of NF-κB and MCP-1 mRNA was detected by real-time quantitative RTPCR. The protein level of NF-κBp65 was detected by Western blot. The concentration of MCP-1 in supernatant was detected by ELISA. Results (1) Compared with Con group, the expression of NF-κB mRNA and MCP-1 mRNA in HG group increased (P <0.05); the relative expression of NF-κBp65 increased (1.04 ± 0.01 vs 0.51 ± 0.15, P 0.05). The level of MCP-1 in the supernatant increased (561.99 ± 23.24 vs 480.73 ± 35.40 pg / ml, P <0.05). (2) The expression of NF-κB mRNA and MCP- -κBp65 protein, the content of MCP-1 in supernatant decreased (P <0.05). Conclusion Metformin can inhibit the expression and synthesis of NF-κB and MCP-1 in MCs, which may be related to their protective effect on the kidney.