中性粒细胞弹性蛋白酶基因mRNA在不同结核分枝杆菌感染人群表达的差异  

Expression of ELANE mRNA in different MTB infected populations

作　　者：樊丽媛 李昆 孙卫国 安红娟 解建辉 张灵霞 FAN Liyuan;LI Kun;SUN Weiguo;AN Hongjuan;XIE Jianhui;ZHANG Lingxia(Luanzhou People's Hospital,Luanzhou,Luanzhou,Hebei 063700,China;Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment,Institute for Tuberculosis Research,the Eighth Medical Center of General Hospital of PLA,Beijing 100091,China)

机构地区：[1]河北省滦州市人民医院,河北滦州063700 [2]解放军总医院第八医学中心,结核病诊疗新技术北京市重点实验室,北京100091

出　　处：《中国热带医学》2025年第2期156-160,共5页China Tropical Medicine

基　　金：国家传染病重大专项(No.2017ZX10201301-007);北京市自然科学基金项目(No.L202027)。

摘　　要：目的 研究中性粒细胞弹性蛋白酶(elastase,neutrophil expresse, ELANE)mRNA在结核分枝杆菌(Mycobacterium tuberculosis, MTB)不同的感染人群外周血单个核细胞(peripheral blood mononuclear cells, PBMCs)中表达特点,分析其作为不同MTB感染人群诊断标志物的可行性。方法 收集2023年2—11月河北省滦州市人民医院活动性结核病患者(active tuberculosis, ATB)、结核分枝杆菌潜伏感染者(latent tuberculosis infection, LTBI)和健康人(healthy control,HC)外周血,分离PBMCs,提取细胞RNA,逆转成cDNA后,采用荧光定量PCR检测各组PBMCs中ELANE基因mRNA水平差异;以结核分枝杆菌H37Rv全菌体裂解液刺激各组PBMCs,提取各组细胞总RNA,同样以荧光定量PCR检测ELANE基因在各组细胞刺激前后的表达差异;分析这种表达差异用于鉴别ATB可行性。结果 ATB患者外周血单个核细胞中ELANE基因表达高于LTBI和健康人群(t=4.550,P<0.001;t=4.540,P<0.001)。ROC分析显示ELANE鉴别诊断ATB与LTBI、ATB与HC的曲线下面积分别为0.802和0.805。H37Rv全菌体裂解液能上调ATB患者PBMCs细胞ELANE基因的表达,刺激前后差异有统计学意义(t=4.717,P<0.001),而LTBI和HC人群在H37Rv菌体裂解液刺激前后,PBMCs中ELANE基因表达均无差异。结论 ATB与LTBI和健康人群相比,其外周血单个核细胞中ELANE基因mRNA的表达水平明显上调;经MTB全菌裂解液抗原刺激后,ATB人群PBMCs中ELANE表达水平升高。ELANE可作为活动性肺结核的诊断标志物,用于鉴别ATB与LTBI和健康人群。Objective To investigate the expression of neutrophil elastase(ELANE)mRNA in peripheral blood mononuclear cells(PBMCs)under different infection states of Mycobacterium tuberculosis(MTB),and analyze the feasibility of ELANE as a diagnostic marker for MTB infection.Methods PBMCs were isolated from peripheral blood of active tuberculosis patients(ATB),latent infection of Mycobacterium tuberculosis(LTBI)and healthy control(HC)Luanzhou People's Hospital,Hebei Province,February-November 2023 and cell RNA was extracted.After reversing into cDNA,the difference of ELANE mRNA level in PBMCs in each group was detected by fluorescence quantitative PCR.The whole cell lysate of Mycobacterium tuberculosis H37Rv was used to stimulate PBMCs in each group,and total RNA was extracted.Fluorescence quantitative PCR was also used to detect the expression differences of ELANE gene before and after cell stimulation in each group;these expression differences were analyzed for identifying the feasibility of ATB.Results The expression of ELANE gene in peripheral blood mononuclear cells of ATB patients was significantly higher than that of LTBI and healthy individuals(t=4.550,P<0.001;t=4.540,P<0.001).ROC analysis showed that the area under the curve for ELANE differential diagnosis of ATB and LTBI,ATB and HC were 0.802 and 0.805,respectively.H37Rv whole cell lysate can upregulate the expression of ELANE gene in PBMCs of ATB patients,with a significant difference before and after stimulation(t=4.717,P<0.001).However,there was no significant difference in ELANE gene expression in PBMCs of LTBI and HC populations before and after H37Rv bacterial lysate stimulation.Conclusion Compared with LTBI and healthy individuals,the expression level of ELANE mRNA in peripheral blood mononuclear cells of ATB was significantly upregulated;After stimulation with MTB whole cell lysate antigen,the expression level of ELANE in PBMCs of ATB population significantly increased.ELANE can serve as a diagnostic biomarker for active pulmonary tuberculosis to different

关 键 词：活动性结核病 潜伏感染 诊断标志物 中性粒细胞弹性蛋白酶 

分 类 号：R521[医药卫生—内科学]