摘要目的 体外观察尿毒症患者血清对人脐静脉内皮细胞(HUVEC)合成单核细胞趋化蛋白1(MCP-1)的影响,探讨诱导血红素氧合酶1(HO-1)高表达对调节MCP-1合成的作用.方法 以含10%尿毒症血清的M199培养基体外培养HUVEC细胞株.采用RT-PCR法检测HUVEC MCP-1 mRNA的表达.用ELISA法检测HUVEC MCP-1蛋白的分泌.然后分别以HO-1诱导剂氯化血红素(hemin)及阻断剂原卟啉锌(ZnPP)预处理HUVEC,应用RT-PER法和免疫组织化学法检测细胞内HO-1及MCP-1 mRNA及蛋白的表达.结果 尿毒症血清可上调细胞MCP-1 mRNA的表达,促进MCP-1蛋白合成,MCP-1蛋白量为对照组的2.95倍.hemin诱导的HO-1高表达可下调尿毒症血清引起的MCP-1基因表达,抑制MCP-1蛋白合成,ZnPP可阻断其作用.30 μmol/L hemin可使HUVEC合成MCP-1蛋白减少34.4%,而hemin与ZnPP共同作用组的MCP-1蛋白量恢复至尿毒症血清组的98.0%.结论 尿毒症血清可诱导HUVEC的MCP-1高表达.促进HO-1高表达时可抑制MCP-1合成及分泌.HO-1有减轻尿毒症环境对内皮细胞功能损伤的作用.

abstractsObjective To investigate the influence of uremic serum on the monocyte chemoattractant protein 1 (MCP-1) expression of human umbilical vascular endothelial cells (HUVECs) in vitro and the effect of up-regulation of berne oxygenase 1 (HO-1) on the synthesis of MCP-1 of HUVECs in uremic milieu. Methods HUVECs were incubated to confluence and then preineubated with heroin and/or protoporphyrin zinc IX (ZnPP)for 6 hours.The cultures were subsequently incubated with M199 cell medium containing 10% serum of healthy people or with medium containing 10% serum of maintenance hemodialysis (MHD) patients.HO-1 protein and mRNA expression was detected by immunohistochemistry and semi-quantitative RT-PCR.MCP-1 mRNA expression was measured by semi-quantitative RT-PCR,and MCP-1 protein was quantified by ELISA. Results Up-regulated expression of MCP-1 mRNA and protein was detected in HUVECs incubated with medium containing 10% serum of MHD patients.The protein synthesis was 2.95 folds of the control.Heroin induced expression of HO-1 mRNA and protein,and concurrently inhibited the up-regulated MCP-1 expression induced by uremic serum.Such effects of heroin could be blocked by ZnPP. Conclusions Uremic serum induces the expression of MCP-1 in HUVECs.Up-regulated expresson of endothelial HO-1 induced by heroin inhibits the enhancement of MCP-1 synthesis.HO-1 may be beneficial to the alleviation of endothelial cell injury in uremic milieu.