miRNA-192调控NOD2在结核分枝杆菌感染中的作用研究

The role of miRNA-192in regulating NOD2in tuberculosis infection

目的探讨miRNA-192调控NOD2在结核分枝杆菌感染中的作用。方法收集2018年1月-2018年12月新疆医科大学第一附属医院确诊为活动性肺结核患者和非结核健康对照者的外周血标本,其中活动性肺结核组93例,健康对照组83例,采用miRanda软件预测NOD23’UTR与miRNA-192是否有结合位点;通过qRT-PCR检测miRNA-192和NOD2 mRNA的相对表达量;通过ELISA检测相关细胞因子IL-6、TNF-α、IFN-γ在血清中的表达情况。结果miRanda软件预测NOD23’UTR中存在miRNA-192的结合位点;qRT-PCR检测miRNA-192和NOD2 mRNA在两组受检者的外周血白细胞中均有表达,其中miRNA-192在活动性肺结核组的表达(5.07±5.66)较健康对照组(0.43±1.13)增加,NOD2 mRNA在活动性肺结核组的表达(1.91±0.12)较健康对照组(1.00±0.63)也增加;ROC曲线分析显示,miRNA-192和NOD2 mRNA肺结核诊断的ROC曲线下面积分别为0.8458(95%CI为0.7997-0.9485)、0.7674(95%CI为0.6674-0.8615),特异性为89.55%和77.94%,敏感度为68.12%和68.06%;活动性肺结核组患者血清IFN-γ、IL-6和TNF-α分别为(36.39±9.24)pg/ml、(62.39±15.37)pg/ml和(77.99±18.82)pg/ml,健康对照组分别为(15.99±4.88)pg/ml、(31.68±7.85)pg/ml和(64.22±20.90)pg/ml,差异均具有统计学意义(P<0.01)。结论结核分枝杆菌感染机体时,高表达的miRNA-192使NOD2 mRNA表达增强,促进相关炎性因子高表达,这对肺结核的发生发展及防治具有临床意义。

Objective To investigate the role of miRNA-192 in regulating NOD2 in Mycobacterium tuberculosis infection. Methods Peripheral blood samples were collected from patients with active pulmonary tuberculosis and healthy controls without tuberculosis in the First Hospital Affiliated with Xinjiang Medical University from January 2018 to December 2018,including 93 patients with active pulmonary tuberculosis and 83 healthy controls.The software miRanda was used to predicted whether NOD2 3’UTR and miRNA-192 have binding sites.The expression of miRNA-192 and NOD2 mRNA was detected using qRT-PCR.The expression of the related cytokines IL-6,TNF-ɑ,and IFN-γin serum was detected using ELISA. Results The software miRanda predicted that an miRNA-192 binding site would be present inNOD2 3’UTR.qRT-PCR revealed miRNA-192 and NOD2 mRNA in peripheral blood leukocytes from both groups.The level of miRNA-192 expression in patients with pulmonary tuberculosis(5.07±5.66)was higher than that in healthy controls(0.43±1.13),and the level of NOD2 mRNA expression in patients with active tuberculosis(1.91±0.12)was also higher than that in healthy controls(1.00±0.63).ROC curve analysis indicated that the AUC for diagnosis of tuberculosis with miRNA-192 was 0.8458(95% CI:0.7997-0.9485)while that for NOD2 mRNA was 0.7674(95% CI:0.6674-0.8615).miRNA-192 had a specificity of 89.55% while NOD2 mRNA had a specificity of 77.94%;miRNA-192 had a sensitivity of 68.12% while NOD2 mRNA had a sensitivity of 68.06%.Serum IFN-γ,IL-6,and TNF-α were 36.39±9.24 pg/ml,62.39±15.37 pg/ml,and 77.99±18.82 pg/ml in patients with active tuberculosis.Serum IFN-γ,IL-6,and TNF-α were 15.99±4.88 pg/ml,31.68±7.85 pg/ml,and 64.22±20.90 pg/ml in healthy controls.The differences were statistically significant(P<0.01).Conclusion When M.tuberculosis infects the body,a high level of miRNA-192 expression increases the expression of NOD2 mRNA and promotes high levels of expression of related inflammatory factors.This finding has clinical significance for the development,progression,and prevention of tuberculosis.