结核病患者外周血白细胞介素-8、信号传导和转录激活因子3表达差异及其辅助诊断价值  

Differential expression analysis of IL-8 and STAT3 in active tuberculosis patients and its value in adjuvant diagnosis

作　　者：安红娟 李昆 张灵霞[1] 朱琰[1] 李改平[1] 史阅韩 孙卫国 AN Hongjuan;LI Kun;ZHANG Lingxia;ZHU Yan;LI Gaiping;SHI Yuehan;SUN Weiguo(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]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》2024年第4期411-416,共6页China Tropical Medicine

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

摘　　要：目的探索耐多药结核病患者(multi-drug resistant tuberculosis,MDR-TB)、药物敏感结核病患者(drug susceptible tuberculosis,DSP-TB)及健康人3组人群外周血白细胞介素-8(interleukin-8,IL-8)、信号传导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)表达差异,为开展新型结核病实验室辅助诊断提供参考依据。方法对3组人群外周血单个核细胞(peripheral blood mononuclear cell,PBMC)mRNA芯片表达谱差异性进行分析;选取MDR-TB患者与DSP-TB患者为研究对象,以健康人为对照组,酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)测定各组血清IL-8浓度,比较组间差异;同时分离3组人群PBMC,收集细胞并提取总RNA,分别通过实时荧光定量PCR(quantitative real-time PCR,qPCR)技术分析各组IL-8、STAT3 mRNA表达情况;以蛋白免疫印迹(Western Blot)实验比较各组STAT3蛋白表达差异。结果MDR-TB和DSP-TB 2组人群血清IL-8浓度明显高于健康对照组,差异有统计学意义(P<0.05);MDR-TB组和DSP-TB组人群外周血PBMC中IL-8基因mRNA的相对表达量明显高于健康对照组,差异有统计学意义(P<0.05);qPCR与Western Blot实验结果均显示,健康组、DSP-TB组和MDR-TB组STAT3基因表达依次上调,组间差异有有统计学意义(P<0.05),实验所得结果与mRNA芯片结果一致。结论IL-8在活动性结核病(active tuberculosis,ATB)患者体内表达显著升高,可以作为标志物区分ATB与健康人群;STAT3浓度的变化不仅作为ATB的诊断参考,还可以鉴别DSP-TB与MDR-TB人群。IL-8、STAT3联合检测有望成为不同结核病患者的临床辅助诊断依据。Objective To compare the differences in the expression of interleukin-8(IL-8)and signal transducer and activator of transcription 3(STAT3)among three groups of individuals:patients with multi-drug resistant tuberculosis(MDRTB),patients with drug-susceptible tuberculosis(DSP-TB),and healthy controls.By analyzing the differences between these groups,the study seeks to provide a reference basis for clinical auxiliary diagnosis of new type tuberculosis.Methods The differences in mRNA expression profiles of chip from peripheral blood mononuclear cells(PBMCs)among three groups were analyzed.MDR-TB and DSP-TB patients were selected as study subjects,with healthy individuals serving as a control group,and the serum IL-8 concentration of each group was measured using an ELISA assay to compare differences between groups.Additionally,PBMCs from all groups were isolated and total RNA was extracted for analysis of IL-8 and STAT3 mRNA expression using quantitative real-time PCR(Q-PCR).Differences in STAT3 protein expression among the groups were also compared by Western blot assay.Results The serum IL-8 concentration in the MDR-TB and DSP-TB groups was significantly higher than that in the healthy control group,with a statistically significant difference(P<0.05).The relative expression level of IL-8 mRNA in PBMCs of the MDR-TB group and DSP-TB group was also significantly higher than that of the healthy control group,with a statistical significance(P<0.05).Both Q-PCR and Western Blot results revealed that the expression of STAT3 in the healthy,DSP-TB and MDR-TB groups sequentially increased,presenting statistically significant differences between the groups(P<0.05),consistent with the mRNA chip results.Conclusions The expression of IL-8 in RPMIactive tuberculosis patients(ATB)is significantly increased,which can be used as a marker to distinguish ATB from healthy populations.The changes of STAT3 concentration not only serve as a reference for the diagnosis of ATB but can also distinguish between DSP-TB and MDR-TB populations.T

关 键 词：结核分枝杆菌 活动性结核病 白细胞介素-8 信号传导和转录激活因子3 

分 类 号：R521[医药卫生—内科学]