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目的探讨肿瘤坏死因子α（ＴＮＦα）对培养的人肾小球内皮细胞（ＨＵＧＥＣ）表达单核细胞趋化蛋白１（ＭＣＰ１）的影响以及ＨＵＧＥＣ的条件培养基对单核细胞（ＭＣ）的趋化作用及抗ＭＣＰ１抗体对单核细胞迁移的影响。方法（１）采用原位杂交技术、免疫细胞化学、细胞ＥＬＩＳＡ法观察ＭＣＰ１基因及蛋白表达。（２）用改良的Ｂｏｙｄｅｎ小室微孔滤膜法测定ＴＮＦα刺激ＨＣＧＥＣ后的条件培养基对ＭＣ的趋化作用及抗ＭＣＰ１抗体对ＭＣ迁移的影响。结果（１）在不加刺激条件下培养的ＨＵＧＥＣ弱表达ＭＣＰ１基因及蛋白，５０ｎｇ／ｍｌＴＮＦα刺激后，６小时即有ＭＣＰ１蛋白表达增强，于１２小时达高峰，不同浓度的ＴＮＦα（２５、５０、１００ｎｇ／ｍｌ）刺激ＨＵＧＥＣ６小时后，与正常对照组相比差异显著（Ｐ＜００１）。（２）ＴＮＦα刺激ＨＵＧＥＣ后的条件培养基对ＭＣ有明显趋化作用，并被抗ＭＣＰ１抗体抑制。结论ＨＵＧＥＣ在ＴＮＦα诱导下，其ＭＣＰ１的表达增强，其条件培养基对ＭＣ有趋化作用，从而可能招引单核细胞迁入内皮下间隙。
Objective To investigate the effect of tumor necrosis factor α (TNFα) on the expression of monocyte chemoattractant protein-1 (MCP1) in cultured human glomerular endothelial cells (HUGEC) and the effect of conditioned medium of HUGEC on monocyte (MC) Chemotaxis and anti-MCP 1 antibody on monocyte migration. Methods (1) The in situ hybridization, immunocytochemistry, cell ELISA assay MCP 1 gene and protein expression. (2) The modified Boyden chamber micropore membrane method was used to determine the chemotactic effect of conditioned media of TNFα-stimulated HCGEC on MC and the effect of anti-MCP1 antibody on MC migration. Results (1) MCP1 gene and protein were weakly expressed in HUGEC without stimulus, MCP1 protein increased at 6 hours and reached the peak at 12 hours after stimulation with 50 ng / ml TNFα. Different concentrations of TNFα 25,50,100ng / ml) stimulated HUGEC for 6 hours, compared with the normal control group, the difference was significant (P <0 01). (2) Conditioned medium conditioned by TNFα stimulated by HUGEC has significant chemotaxis to MC and is inhibited by anti-MCP 1 antibody. Conclusion HUGEC induced by TNFα, the expression of MCP 1 increased, the conditioned medium of MC chemotaxis, which may attract mononuclear cells into the subepithelial space.