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目的研究1,25-二羟维生素D3对胰岛β细胞的免疫保护作用,为胰岛β细胞的免疫保护治疗提供实验依据。方法培养胰岛β细胞株NIT细胞及正常对照外周血单个核细胞,分别与以下4组作用:①(IL-1β+IFN-γ)组,②(IL-1β+IFN-γ+D3)组,③D3组,④对照组(DMEM)。采用酶联免疫吸附分析(ELISA)检测培养上清液中IL-4、IFN-γ水平,用硝酸还原酶法检测上清液NO水平,用化学发光法检测上清液胰岛素水平。结果①IL-1β+IFN-γ+D3刺激的NIT-1细胞与IL-1β+IFN-γ刺激组相比,分泌IL-4水平增高(P<0.01)、IFN-γ减少(P<0.01),IL-4/IFN-γ增高(P<0.01);②IL-1β+IFN-γ+D3刺激的NIT-1细胞与IL-1β+IFN-γ刺激组相比,分泌NO水平减少(P<0.01);③IL-1β+IFN-γ+D3刺激的NIT-1细胞与IL-1β+IFN-γ刺激组相比,胰岛素分泌增加(P<0.05)。结论 1,25-二羟维生素D3可减轻IL-1β+IFN-γ对NIT细胞的损伤作用,保护NIT细胞的胰岛素分泌功能。
Objective To study the protective effect of 1,25-dihydroxyvitamin D3 on islet β cells and provide experimental evidence for the immunoprotection of islet β cells. Methods Cultured NIT cells of islet β cell line and normal control peripheral blood mononuclear cells, respectively, with the following four groups: ① (IL-1β + IFN-γ) group, ② (IL-1β + IFN-γ + D3) ③ D3 group, ④ control group (DMEM). The levels of IL-4 and IFN-γ in culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA). The levels of NO in supernatants were detected by nitrate reductase method and the levels of insulin in supernatants were detected by chemiluminescence. Results ① Compared with IL-1β + IFN-γ stimulation group, IL-4 secretion increased (P <0.01) and IFN-γ decreased in IL-1β + IFN- (P <0.01). (2) Compared with IL-1β + IFN-γ stimulation group, NIT-1 cells stimulated by IL-1β + IFN-γ + D3 decreased NO level (P < 0.01). ③Compared with IL-1β + IFN-γ stimulation group, NIT-1 cells stimulated by IL-1β + IFN-γ + D3 increased insulin secretion (P <0.05). Conclusion 1,25-dihydroxyvitamin D3 can reduce the injury of NIT cells induced by IL-1β + IFN-γ and protect the insulin secretion of NIT cells.