前列腺素E1改善高糖联合肿瘤坏死因子α损伤肾小球系膜细胞株细胞及机制研究

PGE_1 ameliorates injury of rat mesangial cells induced by high glucose and TNF-αand its potential mechanism

目的观察前列腺素E1(PGE1)对高糖联合TNF-α诱导的大鼠肾小球系膜细胞株(HBZY-1)损伤的影响及机制。方法将HBZY-1细胞分为正糖(NG)组、高糖+TNF-α(HT)组、高糖+TNF-α+不同浓度PGE1(HTP1～5)组,采用MTT法测定细胞增殖,ELISA与RT-PCR法测定细胞上清液中单核细胞趋化蛋白-1(MCP-1)、转化生长因子-β1(TGF-β1)以及白介素-6(IL-6)蛋白含量及mRNA表达,细胞免疫荧光法测定核因子-κB p65(NF-κB p65)核转位。结果高糖联合TNF-α促进HBZY-1增殖,增加MCP-1、TGF-β1蛋白与mRNA,以及IL-6mRNA的表达(P<0.05),同时促进NF-κB p65的核转位;1ng/ml的PGE1可抑制上述作用(P<0.05)。结论 PGE1通过抑制NF-κB通路来抑制高糖联合TNF-α诱导的HBZY-1增殖,降低MCP-1、TGF-β1蛋白与基因,以及IL-6基因表达。

Objective To investigate the effects of PGE1 on injury of rat mesangial cells（HBZY-1）induced by high glucose and TNF-α and its possible mechanism.Methods Rat mesangial cells were cultured in the presence of high glucose and TNF-α with different concentrations of PGE1.Cell proliferation was tested by MTT.The protein level of MCP-1,TG-F β1 and I1-6 in cultural supernatants were measured by enzyme linked immunosorbent assay.The mRNA expressions of MCP-1,TGF-β1 and IL-6 were measured by real-time fluorescent quantitative PCR,and NF-κB p65 translocation was detected using immunofluorescence assay.Results High glucose combined with TNF-α obviously promoted HBZY-1 proliferation and increased the protein and mRNA expression of MCP-1 and TGF-β1,and IL-6 mRNA expression （P＜0.05）,and at the same time promoted nuclear transfer of NF-κB p65.And PGE1inhibited above actions （P＜0.05）.Conclusion PGE1 can restrain HBZY-1 proliferation induced by high glucose and TNF-α,and reduce the expression of MCP-1,TGF-β1 and IL6 protein and mRNA via NF-κBpathway.