HupB诱导IL-6 mRNA与miR-144-3p特异性表达及其在结核病诊断中的应用  

HupB induces specific expression of IL-6 mRNA and miR-144-3pand its applications in tuberculosis diagnosis

作　　者：周怀珩 张叶 司马重园 魏毓彤 张其帆 余晓丽[1] 刘晓倩 ZHOU Huaiheng;ZHANG Ye;SIMA Chongyuan;WEI Yutong;ZHANG Qifan;YU Xiaoli;LIU Xiaoqian(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China)

机构地区：[1]武汉轻工大学生命科学与技术学院,武汉430023

出　　处：《武汉轻工大学学报》2025年第1期35-40,78,共7页Journal of Wuhan Polytechnic University

基　　金：武汉市科技局2020年度应用基础前沿科研项目(编号:2020020601012231)。

摘　　要：HupB作为一种结核分枝杆菌(Mycobacterium tuberculosis, Mtb)的核相关蛋白,在Mtb侵染宿主细胞的过程中,对Mtb的存活起关键作用,并同时参与调节宿主的免疫反应。本文探讨基于HupB特异性诱导IL-6 mRNA及miR-144-3p在外周血单个核细胞(Peripheral blood mononuclear cell, PBMC)中的表达及其应用,为结核病筛查、辅助诊断与免疫治疗提供理论依据。选取临床全血样本,用密度梯度离心法收集PBMC,经HupB不同时间诱导后,提取细胞总RNA并逆转录合成cDNA,使用q-PCR检测IL-6 mRNA及miR-144-3p表达水平,并进行ROC曲线分析;利用皮尔森相关系数分析HupB诱导PBMC后IL-6 mRNA和miR-144-3p的相关性。结果显示在HupB诱导PBMC后,IL-6 mRNA表达呈上升趋势,且在4 h时达到峰值(P<0.001),IL-6 mRNA的相对表达量在肺结核(Pulmonary tuberculosis, PTB)患者的PBMC中显著高于非肺结核肺部疾病(Non-pulmonary tuberculosis, nPTB)患者(P<0.001),此时AUC为0.712 8,最佳cut-off值为0.864 1;miR-144-3p表达量呈下降趋势,且在3 h时降幅最大(P<0.001),PTB组的miR-144-3p表达量最低(P<0.001),此时AUC为0.870 0,最佳cut-off值为1.877。皮尔森相关系数分析结果显示,HupB诱导PTB PBMC 3 h后IL-6 mRNA与miR-144-3p之间呈负相关(R=-0.64,P=0.02)。以上结果提示IL-6 mRNA可能成为肺结核与其他非结核肺部疾病的鉴别诊断标志物,miR-144-3p可能在IL-6 mRNA介导的结核免疫应答中发挥作用,且miR-144-3p有望用于肺结核疾病的筛查,具有成为疾病治疗靶点的潜力。As a nuclear-associated protein of Mycobacterium tuberculosis,HupB plays an important role in Mtb survival after Mtb infection and regulates the host's immune response.The research paper is based on the expression and immunological application of IL-6 mRNA and miR-144-3pspecifically induced by HupB in PBMC,providing a theoretical basis for tuberculosis screening,auxiliary diagnosis and immunotherapy.Clinical whole blood samples were selected,PBMC were collected by density gradient centrifugation,and after HupB induction at different times,total cellular RNA was extracted and reverse transcribed to synthesize cDNA.Q-PCR was used to detect the expression levels of IL-6 mRNA and miR-144-3p,and ROC curve analysis was performed.Pearson correlation coefficient was used to analyze the correlation between IL-6 mRNA and miR-144-3p after HupB-induced PBMC.The results showed that after HupB induced PBMC,IL-6 mRNA expression showed an upward trend and peaked at 4 h(P<0.001).For patients with PTB the relative expression of IL-6 mRNA in their PBMC was significantly higher than that in PBMC for patients with nPTB(P<0.001).At this time,AUC is 0.7128,the best cut-off value is 0.8641;the expression of miR-144-3p shows a downward trend,and the largest decrease occurs at 3 h(P<0.001).The expression level of PTB miR-144-3p in PTB group was the lowest(P<0.001),and this time,AUC is 0.8700 and the optimal cut-off value of 1.877.Pearson correlation coefficient analysis showed a negative correlation between IL-6 mRNA and miR-144-3p after HupB-induced PTB PBMC for 3 h(R=-0.64,P=0.02).It is proposed that IL-6 mRNA could serve as a differential diagnostic marker for tuberculosis as well as for other non-tuberculous lung diseases.Furthermore,miR-144-3p is anticipated to be utilized for the screening of tuberculosis and may be involved in the IL-6 mRNA-mediated immune response to tuberculosis,which may provide a potential therapeutic target.

关 键 词：结核病 HupB PBMC IL-6 mRNA miR-144-3p 肺结核病诊断 

分 类 号：R392.12[医药卫生—免疫学]