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目的:探讨CD4+CD25+调节性T细胞(Tregs)对氧化型低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVECs)炎性因子表达的影响。方法:磁性细胞分离器(MACS)分离CD4+CD25+T细胞及CD4+CD25-T细胞。在ox-LDL作用下,将HUVECs分别与anti-CD3mAb激活的CD4+CD25+T细胞、CD4+CD25-T细胞共培养24h。分别应用流式细胞术、ELISA、real-timePCR测定Tregs对ox-LDL诱导损伤HUVECs炎性因子VCAM-1、MCP-1、IL-6表达的影响。应用凝胶电泳迁移率实验(EMSA)方法观察Tregs对ox-LDL诱导损伤HUVECsNF-κB激活的影响。结果:与对照组比较,Tregs细胞可显著抑制ox-LDL诱导损伤HUVECs炎性因子(VCAM-1、MCP-1、IL-6)及NF-κB的结合活性。结论:Tregs细胞可显著抑制ox-LDL诱导损伤HUVECs炎性因子的表达,其作用机制可能为下调了NF-κB的结合活性。
Objective: To investigate the effect of CD4 + CD25 + regulatory T cells (Tregs) on the expression of inflammatory cytokines in human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL). Methods: CD4 + CD25 + T cells and CD4 + CD25-T cells were isolated by magnetic cell separator (MACS). Under the action of ox-LDL, HUVECs were co-cultured with CD4 + CD25 + T cells and CD4 + CD25-T cells activated by anti-CD3 mAb for 24 h respectively. The effects of Tregs on the expression of inflammatory factors VCAM-1, MCP-1 and IL-6 in HUVECs induced by ox-LDL were detected by flow cytometry, ELISA and real-time PCR respectively. The effect of Tregs on the activation of NF-κB induced by ox-LDL in HUVECs was observed by electrophoretic mobility shift assay (EMSA). Results: Compared with the control group, Tregs cells could significantly inhibit the binding activity of inflammatory factor (VCAM-1, MCP-1, IL-6) and NF-κB induced by ox-LDL in HUVECs. CONCLUSION: Tregs cells can significantly inhibit the expression of inflammatory cytokines induced by ox-LDL in HUVECs. The possible mechanism is that Tregs can down-regulate the binding activity of NF-κB.