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目的探讨高糖环境下人脐静脉内皮细胞株(HUVEC)表面协同刺激分子OX40L表达变化以及对T淋巴细胞生物学行为的影响。方法分别以5.5、16.7、33.3mmol/L浓度葡萄糖干预HUVEC为干预组,单纯DMEM培养为对照组,孵育72h后应用流式细胞术检测细胞表面OX40L的表达变化。16.7mmol/L浓度葡萄糖干预的HUVEC与人T细胞共培养,分为六组:A组(T细胞)、B组(激发型鼠抗人CD3单抗+T细胞)、C组(CD3单抗+T细胞+HUVEC)、D组(CD3单抗+T细胞+HUVEC+阻断型抗人OX40L抗体)、E组(CD3单抗+T细胞+HUVEC+16.7mmol/L葡萄糖)、F组(CD3单抗+T细胞+HUVEC+阻断型抗人OX40L抗体+16.7mmol/L葡萄糖),72h后用细胞增殖和细胞计数试剂盒(CCK-8)检测T细胞增殖。结果不同浓度葡萄糖作用细胞72h,细胞表面OX40L的表达较对照组显著升高(P<0.05),其效应呈现一定的浓度依赖性。高糖作用后的HUVEC促进T细胞增殖能力增强(P<0.05),抗OX40L单抗阻断后促进作用减弱(P<0.05)。结论高糖能有效上调HUVEC表面负性协同刺激分子OX40L表达,促进T细胞增殖。
Objective To investigate the changes of OX40L expression in human umbilical vein endothelial cell line (HUVEC) under high glucose and its effect on the biological behavior of T lymphocytes. Methods HUVECs were treated with glucose at concentrations of 5.5, 16.7 and 33.3 mmol / L, respectively. The cultured cells were cultured in DMEM alone. The expression of OX40L on the cell surface was detected by flow cytometry 72 h after incubation. HUVECs interfered with 16.7mmol / L glucose were co-cultured with human T cells and divided into six groups: group A (T cells), group B (stimulated mouse anti-human CD3 monoclonal antibody + T cells), group C (CD3 monoclonal antibody + T cell + HUVEC + blocking type anti-human OX40L antibody), group E (CD3 monoclonal antibody + T cell + HUVEC + 16.7 mmol / L glucose), group F M + T cells + HUVEC + blocking anti-human OX40L antibody + 16.7 mmol / L glucose) and T cell proliferation was measured 72h later using the Cell Proliferation and Cell Counting Kit (CCK-8). Results The expression of OX40L on the cell surface was significantly increased at different concentrations of glucose for 72 hours (P <0.05), and the effect was dose-dependent. HUVEC after high glucose treatment enhanced the ability of T cell proliferation (P <0.05), and decreased the anti-OX40L mAb (P <0.05). Conclusion High glucose can effectively up-regulate the expression of negative co-stimulatory molecule OX40L on HUVEC surface and promote T cell proliferation.