差异表达的miR-181a-5p、miR-141-3p在青海藏族肺结核诊断中作用

Role of differential expression of miR-181a-5p and miR-141-3p in diagnosis of tuberculosis among Tibetans in Qinghai province

目的探讨青海藏族肺结核患者活动期、潜伏感染期及健康对照组间循环血中hsa-miR-181a-5p、hsa-miR-141-3p的表达情况,判定其在青海藏族肺结核活动期、潜伏期的表达差异。方法分别采集青海地区藏族肺结核患者活动期、潜伏期及同期体检健康者血液样本。提取各组miRNA,用Human miFinder miscript miRNA PCR芯片进行qRT-PCR检测分析,最后进行验证miRNA的表达情况。结果基因芯片检测结果提示hsa-miR-181a-5p在结核组中肺结核活动期患者与健康者比较FC=0.1874,P=0.0001;潜伏期患者与健康者比较为FC=0.2243,P=0.0057,活动期肺结核患者与潜伏期患者相比FC=0.8844,P=0.759224。hsa-miR-141-3p在肺结核组与健康对照组比较FC=0.1293,P=0.0355;潜伏感染组为FC=0.3117,P=0.17194,活动期肺结核患者与潜伏期患者相比FC=2.4113,P=0.958698。结论基因芯片检测结果显示青海地区藏族肺结核患活动期及潜伏期循环血中miRNA的表达较健康对照组具有差异性,hsa-miR-181a-5p、hsa-miR-141-3p在结核分支杆菌感染后的机体中呈低表达,其中hsa-miR-181a-5p在肺结核活动期与潜伏期均呈低表达,但hsa-miR-141-3p只在活动期肺结核患者中呈低表达,因此这两者miRNA联合检测作为肺结核诊断、分期的指标,有望成为诊断高原地区藏族肺结核患者特异表达的分子标志物。

Objective To investigate the expression levels of hsa-miR-181a-5p and hsa-miR-141-3p in the circulating blood of Tibetan patients with tuberculosis in the active stage and latent infection stage and healthy volunteers in order to determine the role of their differential expression in diagnosis of tuberculosis in Tibetans.Methods The blood samples were collected respectively from Tibetan tuberculosis patients in Qinghai province during the active period and the latent period and the healthy volunteers from physical examinations at the same time.After miRNA was extracted from each group,human miFinder miscript miRNA PCR chip and quantitative real-time polymerase chain reaction(qRT-PCR)were used for detection and analysis,and miRNA expression was verified.Results The results of gene chip detection found that FC=0.1874 and P=0.0001 for the hsa-mir-181a-5p in the tuberculosis patients in the active phase compared with the healthy volunteers,FC=0.2243 and P=0.0057 for the hsa-mir-181a-5p in the tuberculosis patients in the latent stage compared with the healthy volunteers,and FC=0.8844 and P=0.759224 for the hsa-mir-181a-5p in the tuberculosis patients in the active stage compared with that of the tuberculosis patients in the latent stage.The results of gene chip detection found that FC=0.1293 and P=0.0355 for the hsa-miR-141-3p in the tuberculosis group compared with the healthy control group,FC=0.3117 and P=0.17194 in the latent infection group compared with the healthy control group,and FC=2.4113 and P=0.958698 in the tuberculosis group compared with the latent infection group.Conclusion The testing results of gene chip in Qinghai Tibetan region in the active pulmonary tuberculosis and in the incubation period and the expression of miRNA in the circulating blood have significant differences compared with the healthy control group,including the lower expression levels of hsa-miR-181a-5p and hsa-miR-141-3p in the patients with mycobacterium tuberculosis infection.And the hsa-miR-181a-5p shows lower expression levels in the active and latent stages of pulmonary tuberculosis,but the hsa-miR-141-3p only shows low expression levels in the active stage of pulmonary tuberculosis patients.Therefore,the two kinds of miRNA as indices of tuberculosis diagnosing and staging are expected to be molecular markers for a specific expression of Tibetan tuberculosis patients in the plateau regions.