摘要
目的:研究1,25-二羟维生素D3(DHVD3)作用下脊柱结核患者外周血单个核细胞(PBMC)中一氧化氮(NO)及诱导型一氧化氮合酶(iNOS)含量变化,探讨其对脊柱结核患者的免疫调节作用。方法:患者PBMC分为试验组(加入DHVD3)和对照组(加入培养基),接种卡介苗后培养;采用硝酸还原酶法测定不同时间点(0、3、6、9、12d)试验组和对照组培养上清液中NO的相对吸光度值,分析其变化规律;于第6天收获2组培养细胞,提取细胞总RNA,逆转录后,定量聚合酶链反应(PCR)扩增,对目的基因iNOS及参照基因3-磷酸甘油醛脱氢酶的mRNA进行分析,用相对定量方法2-ΔΔCt比较其差异。结果:细胞培养第6天,试验组NO的相对吸光度值和iNOS的mRNA含量均高于对照组(P<0.05)。结论:DHVD3可使脊柱结核患者PBMC内iNOS的含量增加,进而促进NO的分泌,从而增强单个核细胞对结核杆菌的吞噬作用。
OBJECTIVE ： To study the effects of i, 25- droxyvitamin Da on nitric oxide and nitric oxide synthase in peripheral blood mononuclear cells （PBMC） in patients with spinal tuberculosis and to explore its immunoregulation function. METHODS： Patients＇ PBMC were assigned to test group in which DHVDa was added or control group in which culture medium was added, then cultured after BCG vaccine inoculation. Cultured supernatant fluid from isolated PBMC was collected at 0, 3, 6, 9, 12 days respectively for measuring of relative optical density （OD） of NO by nitric oxide kit. Total RNA ex- tracted from the PBMC was reverse transcribed to cDNA and amplificated by fluorecent real - time quantitative PCR on the sixth day; mRNA of target gene iNOS and reference gene 3 - GAPD were analyzed and compared by relative quantitative method 2^-△△Ct, RESULTS： OD value of NO and mRNA of iNOS in test group were all higher than in control group （P〈0.05） on the sixth day. CONCLUSION. 1,25- droxyvitamin Da can up - regulate iNOS in PBMC in patients with spinal tuberculosis so as to promote the secretion of NO and enhance phagocytosis of PBMC in tubercle bacillus.
出处
《中国药房》
CAS
CSCD
北大核心
2008年第19期1474-1476,共3页
China Pharmacy
