拉莫三嗪诱发皮肤不良反应与TCRBV基因表达的相关性研究

发布时间：2018-03-17 05:01

  本文选题：拉莫三嗪　切入点：皮肤不良反应　出处：《宁夏医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的研究拉莫三嗪(LTG)激活T细胞介导免疫应答而诱发皮肤不良反应(cADRs)的可能致病机制。分析在LTG-cADRs患者与LTG耐受者外周血单个核细胞(PBMCs)中,TCRBV基因表达的差异。探讨LTG-cADRs与TCRBV基因表达的相关性。资料与方法收集10例cADRs已治愈达6周以上的LTG-cADRs患者作为病例组,依据病例对照研究的方法按1:1匹配纳入10例LTG耐受者作为对照组,分离病例组与对照组患者PBMCs,给予LTG体外刺激并培养,采用ELISA法测定病例组与对照组患者PBMCs分泌IFN-γ、IL-5及TNF-β的情况,运用实时荧光定量逆转录PCR技术(RT-qPCR)的SYBR GreenⅠ荧光染料法检测TCRBV基因的表达。用两独立样本t(或t')检验比较两组IFN-γ、IL-5及TNF-β的浓度差异。采用2-ΔΔCt相对定量法计算在病例组与对照组中,TCRBV基因的相对表达量。用Mann-Whitney U检验分析两组TCRBV基因相对表达量的差异。结果1、以终浓度为25μg/ml的LTG分别刺激病例组与对照组的PBMCs,发现病例组细胞上清液中IFN-γ浓度(1.06±0.09ng/ml)明显高于对照组(0.86±0.16ng/ml),差异有统计学意义(P0.05)。病例组细胞上清液中IL-5和TNF-β的浓度(0.73±0.06ng/ml,0.69±0.05ng/ml)均略高于对照组(0.70±0.07ng/ml,0.65±0.05ng/ml),差异无统计学意义(P0.05)。2、病例组与对照组TCRBV基因表达存在差异,TCRBV11、TCRBV15、TCRBV17、TCRBV18、TCRBV19在病例组中的相对表达量均较对照组高(P0.05)。结论1、LTG可在体外激活LTG-cADRs患者的PBMCs分泌IFN-γ,提示T细胞介导的免疫反应可能参与了LTG-cADRs的致病机制。2、LTG-cADRs可能与TCRBV11、TCRBV15、TCRBV17、TCRBV18、TCRBV19基因的表达相关。
[Abstract]:Objective to study the possible pathogenesis of LTG activation of T cell mediated immune response and induce skin adverse reaction (ADRs), and to analyze the difference of TCR BV gene expression in peripheral blood mononuclear cells (PBMCs) of patients with LTG-cADRs and LTG tolerant patients, and to explore the possible mechanism of TCR BV gene expression in peripheral blood mononuclear cells (PBMCs) of patients with LTG-cADRs and LTG tolerant patients. Data and methods Ten patients with LTG-cADRs who had been cured of cADRs for more than 6 weeks were collected as the case group. According to the method of case-control study, 10 patients with LTG tolerance were selected as control group according to 1: 1 match. PBMCs were stimulated and cultured with LTG in vitro. The secretion of IFN- 纬 IL-5 and TNF- 尾 by PBMCs in case group and control group was measured by ELISA method. The expression of TCRBV gene was detected by real-time fluorescence quantitative reverse transcriptase PCR (RT-qPCR) SYBR Green 鈪，

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