血红素氧合酶1对尿毒症环境脐静脉内皮细胞单核细胞趋化蛋白1表达的影响

Impact of heme oxygenase 1 on monocyte chemoattractant protein 1 expression of human umbilical vascular endothelial cell in uremic milieu

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-11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—PCR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。30μmol／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。

Objective 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 heme oxygenase 1 （HO-1） on the synthesis of MCP-1 of HUVECs in uremic milieu. Methods HUVECs were incubated to confluence and then preincubated with hemin 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. Hemin induced expression of HO-1 mRNA and protein, and concurrently inhibited the up-regulated MCP-1 expression induced by uremic serum. Such effects of hemin 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 hemin inhibits the enhancement of MCP-1 synthesis. HO-1 may be beneficial to the alleviation of endothelial cell injury in uremic milieu.