lncRNA在结核病患者外周血的差异性表达

Differences in expression of lncRNA in peripheral blood from patients with tuberculosis

目的比较活动性肺结核患者、潜伏感染者与健康对照者外周血结核分枝杆菌特异性抗原刺激前后lncRNA1和lncRNA2的差异性表达,探讨其作为诊断结核分枝杆菌感染生物标志物的可能性。方法收集活动性肺结核患者、潜伏感染者和健康对照者全血,分别在特异性抗原刺激前后提取RNA,利用实时荧光定量PCR方法检测lncRNA1和lncRNA2的mRNA表达水平。应用ELISA检测血浆中IFN-γ和IL-2含量。结果结核分枝杆菌特异性抗原刺激前,结核病组、潜伏感染组全血lncRNA1相对表达量较健康对照组分别下降32%和64%;lncRNA2结核病组相对表达量是健康对照组的6.10倍;抗原刺激后,结核病组、潜伏感染组lncRNA1相对表达量较健康对照组分别下降33%和77%,lncRNA2潜伏感染组相对表达量较健康对照组下降84%。与刺激前相比,抗原刺激后健康对照组、结核病组lncRNA1相对表达量分别提高1.80倍和2.80倍,lncRNA2健康对照组、潜伏感染组分别提高3.50倍和1.86倍(P<0.05)。健康组与潜伏感染组、结核病组与潜伏感染组、健康组与结核病组lncRNA1ROC曲线下面积(AUC)分别为0.700、0.752和0.400,lncRNA2的AUC分别为0.702、0.465和0.762。在结核特异性蛋白刺激前,结核病组IFN-γ、IL-2含量均高于健康组(P<0.05),刺激后,结核病组及潜伏感染组与健康组相比较、潜伏感染组与结核病组相比较IFN-γ均增高(P<0.05)。与刺激前相比,刺激后3组IFN-γ、IL-2诱导表达量均显著增高(P<0.05)。结论活动性结核病、潜伏感染和健康对照者全血结核特异性抗原刺激前后lncRNA1和lncRNA2均出现差异性表达,结核病及潜伏感染者血浆IFN-γ含量增高。lncRNA1和lncRNA2可能是诊断肺结核感染的潜在生物标志物。

Objectives To compare the levels of lncRNA1 and lncRNA2 expression in the peripheral blood of patients with active tuberculosis and patients with a latent tuberculosis infection to levels in healthy controls before and after stim- ulation with Mtb-specific antigens and to explore the use of levels of mRNA expression as possible biomarkers for the di- agnosis of TB. Methods Before and after stimulation with Mtb specific antigens, the peripheral blood of patients with active pulmonary tuberculosis （TB）, a latent tuberculosis infection （LTBI）, and healthy controls was collected and RNA was extracted. The expression of lncRNA1 and lncRNA2 mRNA was detected with real-time quantitative PCR. ELISA was used to detect 1FN-Y and 1L-2 levels in plasma. Results Before stimulation with Mtb-speeific antigens, the relative level of lncRNA1 expression in patients with TB decreased 32%and that in patients with an LTBI decreased 64% in com parison to healthy controls. The relative level of lncRNA2 expression in patients with TB was 6.10 times that in healthy controls. After stimulation with Mtb-speeific antigens, the relative level of lncRNA1 expression in patients with TB de creased 33% and that in patients with an LTBI decreased 77% in comparison to healthy controls. The relative level of ln- cRNA2 expression in patients with TB decreased 84% in comparison to that in healthy controls. After stimulation with Mtb-speeific antigens, the relative level of lneRNA1 expression in healthy controls was 1.80 times that in patients not simulated with those antigens and the relative level of lneRNA1 expression in TB was 2.80 times that in patients not sim ulated with those antigens. The relative level of lneRNA2 expression in healthy controls was 3.50 times that in patients not simulated with antigens and the relative level of lneRNA2 expression in LTBI was 1.86 times that in patients not sire-ulated with antigens （P〈0.05）. Receiver operating characteristic （ROC） curve analysis of lncRNA1 in healthy controls and patients with a LTBI indicated that that the area under the curve （AUC） was 0. 700. ROC curve analysis of lncRNA1 in patients with TB and patients with an LTBI indicated that the AUC was 0. 752. ROC curve analysis of lncRNA1 in healthy controls and patients with TB indicated that that the AUC was 0. 400. ROC curve analysis of lncRNA in healthy controls and patients with a LTBI indicated that that the area under the curve （AUC） was 0. 702. ROC curve analysis of lncRNA2 in patients with TB and patients with an LTBI indicated that the AUC was 0. 465. ROC curve analysis of ln- cRNA2 in healthy controls and patients with TB indicated that that the AUC was 0. 762. Levels of IFN-Y and IL-2 in pa- tients with TB were higher before stimulation with Mtb specific antigens than those in healthy controls （P〈0.05）. After stimulation with Mtb-specific antigens, levels of IFN-Y were compared in patients with TB, patients with an LTBI, and healthy controls. Levels of IFN -Y were higher in patients with an LTBI and in patients with TB （P〈0.05）. Stimulation with Mtb-specific antigens induced significantly higher levels of IFN 7 and IL-2 expression in all three groups （P〈0.05）. Conclusion Significant differences in levels of lncRNA1 and lneRNA2 expression were noted in peripheral blood from patients with TB, patients with an LTBI, and healthy controls before and after stimulation with Mtb specific antigens. Levels of IFN-Y in plasma were higher in patients with active pulmonary TB and an LTBI. lncRNA1 and lncRNA2 may be used as potential biomarkers for the diagnosis of TB.