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[目的]研究1,25-(OH)2D3对结核患者外周血单个核细胞(PBMC)在吞噬结核杆菌卡介苗后的一氧化氮(NO)及诱导型一氧化氮合酶(iNOS)的变化,探讨1,25-(OH)2D3对结核患者的免疫调节作用。[方法]结核患者PBMC经分离培养后,硝酸还原酶法测定不同时间点实验组和对照组培养上清液NO的相对吸光度(OD)值,分析它在不同时间点的变化;于d6收获两组培养的细胞,提取细胞总RNA,逆转录成cDNA,定量聚合酶链反应(PCR)扩增,对目的基因iNOS及参照基因GAPDH的mRNA进行分析,用相对定量方法2-△△Ct比较其差异。[结果]在细胞培养d6,实验组NO的相对OD值为0.045±0.011,高于对照组(P﹤0.05);实验组iNOS的mRNA含量也明显高于对照组,差异有统计学意义(P﹤0.05)。[结论]1,25-(OH)2D3可使结核患者PBMC内iNOS的含量增加,进而促进NO的分泌,增强单个核细胞对结核杆菌的吞噬。
[Objective] To investigate the changes of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) after 1,25- (OH) 2D3 binding to peripheral blood mononuclear cells (PBMCs) of tuberculosis patients after phagocytosis of Mycobacterium tuberculosis, To investigate the immunomodulatory effects of 1,25- (OH) 2D3 on patients with tuberculosis. [Method] PBMC from patients with tuberculosis were isolated and cultured. The relative absorbance (OD) of NO in culture supernatant of experimental group and control group at different time points was determined by nitrate reductase method. The changes of NO at different time points were analyzed. The total RNA of the cells was extracted and reverse transcribed into cDNA. The mRNA of iNOS gene and reference gene GAPDH was analyzed by quantitative polymerase chain reaction (PCR). The relative quantification method difference. [Results] The relative OD value of NO in experimental group was 0.045 ± 0.011, which was higher than that in control group (P <0.05). The mRNA level of iNOS in experimental group was also significantly higher than that in control group (P <0.05). [Conclusion] 1,25- (OH) 2D3 can increase the content of iNOS in PBMC of tuberculosis patients, and then promote the secretion of NO and enhance the phagocytosis of Mycobacterium tuberculosis by mononuclear cells.