痰菌阴性活动性肺结核患者CD4^+ T细胞的miR-29及其靶基因IFN-γ表达水平的诊断价值研究

Study on the value of CD4^+ T cell miR-29 family and target gene IFN-γ in the diagnosis of sputum negative pulmonary tuberculosis

目的探讨痰菌阴性活动性肺结核患者的CD4^+T细胞的miR-29及其靶基因IFN-γ的表达在痰菌阴性活动性肺结核患者诊断中的应用价值。方法采取便利抽样法,选取2017年1—12月在浙医二院长兴院区治疗痰菌阴性活动性肺结核患者60例(痰菌阴性组),痰菌阳性活动性肺结核患者60例(痰菌阳性组),潜伏结核感染者35例(潜伏感染组),选取同期体检42名健康人为对照组。分离纯化4组受试者外周血中的CD4^+T细胞;用核酸杂交与荧光定量PCR检测CD4^+T细胞内miR-29a、miR-29b和miR-29c的表达;用定量PCR检测miR-29的靶基因IFN-γ的表达。结果 miR-29a、miR-29b和miR-29c在痰菌阴性组、痰菌阳性组、潜伏感染组和对照组CD4^+T细胞中的表达水平均不同(P<0.05);miR-29b和miR-29c在痰菌阴性组、痰菌阳性组的表达高于对照组,miR-29a、miR-29b和miR-29c在痰菌阴性组、痰菌阳性组的表达均低于潜伏感染组,差异均有统计学意义(均P<0.05);IFN-γ在4组之间的表达水平亦不同(均P<0.05)。结论 CD4^+T细胞内miR-29a、miR-29b及miR-29c在痰菌阴性活动性肺结核发病过程中可能起到重要作用,这为结核病的诊治提供潜在的靶标和新的策略。

Objective To investigate the application of the expression of the miR-29 family of CD4 ＋ T cells and the ex- pression of target gene IFN-γ in the diagnosis of sputum negative pulmonary tuberculosis. Methods From January to December, 2017, using convenient sampling methods, in our hospital treatment of patients with 60 cases of sputum nega- tive pulmonary tuberculosis （negative group） , 60 cases of sputum positive pulmonary tuberculosis （positive group） , 35 cases of latent tuberculosis infection （infection group） were selected in our hospital physical examination and 42 heahhy people for the control group were selected at the same time. CD4＋ T cells from peripheral blood of 4 groups were separa- ted and purified. The expressions of miR-29a, miR-29b and miR-29c in CD4 ＋ T cells were detected by nucleic acid hy- bridization and fluorescence quantitative PCR, and the expression of miR-29 target gene IFN-γ was detected by quantita- tive PCR. Results The expression level of miR-29a, miR-29b and miR-29c in the negative group, positive group, la- tent infection group and control group in CD4＋ T cells were different （ P 〈 0.05 ）. The expression of miR-29b and miR- 29c in negative group and positive group were higher than that in the control group, and miR-29a, miR-29b and miR-29c in the negative group, positive group were lower than the latent infection group （ P 〈 0.05 ） ; the expression level of target gene IFN-γ in the 4 groups were also different （ P 〈 0.05 ）. Conclusion The miR-29a, miR-29b and miR-29c in CD4 ＋ T cells may play an important role in the pathogenesis of phlegm negative active tuberculosis, which provides a potential target and a new strategy for the diagnosis and treatment of tuberculosis.