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关键词：肾小球；内皮细胞；单核细胞；单核细胞趋化蛋白-1
【摘要 】 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(MCP-1)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MCP-1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MCP-1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MCP-1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MCP-1基因及蛋白，50ng/ml TNFα刺激后，6小时即有MCP-1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<0.01)。(2)TNFα刺激HUGEC后的条件培养基对MC有明显趋化作用，并被抗MCP-1抗体抑制。结论 HUGEC在TNFα诱导下，其MCP-1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。
Expression of monocyte chemoattractant protein-1 in glomerular endothelial cells
DING Hanlu* , ZHU Miaozhen, LUO Xiangdong ,et al. * Department of Nephrology, Daping Hospital, Third Military Medical University, Chongqing 400042
【Abstract 】 Objective Tostudy the expression of monocyte chemoattractant protein-1 (MCP-1) in cultured humanglomerular endothelial cell (HUGEC).Methods The effect of TNFα on theexpression of MCP-1 mRNA and protein by HUGEC was observed with in situ hybridization,immunocytochemistry, cell ELISA analysis . The monocyte migration induced by the mediaconditioned by cultured HUGEC was assayed by micropore filter method using modified BoydenChamber. Influence of anti-MCP-1 antibody on monocyte migration was observed as well.Results mRNAand protein of MCP-1 were only basically expressed on HUGEC at very low levels in thecontrol groups without stimulation. After stimulation by TNFα (50 ng/ml), however,their expression was markedly upregulated from the 6th hour. The maximal proteinexpression of MCP-1 was present at the 12th hour. Compared with control groups, MCP-1protein expression was significantly increased after the stimulation at differentconcentration of TNFα (25,50,100ng/ml) stimulation (P<0.01). HUGEC stimulated by TNFαproduced a factor that was markedly chemotactic for monocytes , and the chemotacticactivity was inhibited by anti -MCP-1 antibody.Conclusion HUGECstimulated by TNFα may highly express MCP-1, and produce chemotactic factors formonocytes.
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