MicroRNA的生物表达与结核病诊断和治疗监测的相关性及其对结核病易感基因表达调控的研究

Study on the correlation of MicroRNA expression with diagnosis and treatment surveillance of tuberculosis and treatment monitoring and its regulation of the expression of tuberculosis susceptibility gene

目的探究微小核糖核酸(MicroRNA)对结核病诊断和治疗监测及易感基因表达调控的意义。方法选取136例肺结核患者作为肺结核组,另选择142例健康体检者作为对照组。以实时荧光定量PCR(qRT-PCR)法检测两组血清hsa-miR-20b、CUL4A mRNA、GBP5 mRNA水平;检测干扰素调节因子8基因(IRF8)、循环换血RNA-103571(hsa-circRNA-103571)、γ-干扰素(IFN-γ)水平。比较两组血清hsa-miR-20b、GBP5 mRNA、CUL4A mRNA的表达量;比较两组血清IRF8、hsa-ciceRNA-103571、IFN-γ水平;分析肺结核患者hsa-miR-20b与IRF8、hsa-circRNA-103571、IFN-γ的相关性;回归分析肺结核发生的影响因素;分析血清hsa-miR-20b对肺结核的诊断效能。结果肺结核组患者血清hsa-miR-20b、GBP5 mRNA、CUL4A mRNA的表达水平分别为(3.35±0.20)、(2.94±0.23)、(0.41±0.15),对照组分别为(1.01±0.23)、(1.02±0.13)、(1.03±0.11);与对照组相比,肺结核组患者血清hsa-miR-20b、GBP5 mRNA的表达水平均升高,CUL4A mRNA的表达水平降低,差异均有统计学意义(P<0.05)。肺结核组患者血清IRF8(389.39±46.85)pg/ml、hsa-circRNA-103571(22.48±5.06)ng/L均高于对照组的(327.62±41.76)pg/ml、(11.36±3.17)ng/L,IFN-γ(5.58±1.36)pg/ml明显低于对照组的(12.72±4.78)ng/L,差异有统计学意义(P<0.05)。Pearson相关性分析显示,肺结核患者血清hsa-miR-20b与IRF8、hsa-circRNA-103571水平呈正相关(r=0.585、0.539,P<0.05),与IFN-γ水平呈负相关(r=-0.485,P<0.05)。以hsa-miR-20b、IRF8、hsa-circRNA-103571、IFN-γ水平为自变量,以肺结核是否发生为因变量,行Logistic回归分析,结果显示hsa-miR-20b、IRF8、hsa-circRNA-103571是肺结核发生的危险因素(P<0.05),IFN-γ是肺结核发生的保护因素(P<0.05)。血清hsa-miR-20b水平对肺结核发生诊断的曲线下面积(AUC)为0.898,截断值为1.44,其灵敏度为83.82%,特异度为88.03%。结论肺结核患者血清hsa-miR-20b表达上调,hsa-miR-20b与炎症相关因子具有一定相关性,其可能与炎症因子共同影响肺结核疾病病变过程。

Objective To explore the significance of micro-ribonucleic acid(MicroRNA)in diagnosis and treatment surveillance and susceptibility regulation of gene expression of tuberculosis patients.Methods A total of 136 patients with pulmonary tuberculosis were selected as the pulmonary tuberculosis group,and another 142 healthy physical examinees were selected as the control group.The serum levels of hsa-miR-20b,CUL4A mRNA and GBP5 mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR).The interferon regulatory factor 8(IRF8),hsa-circRNA-103571 and IFN-γwere detected.The expression serum hsa-miR-20b,GBP5 mRNA and CUL4A mRNA were compared between the two groups;the serum levels of IRF8,hsa-ciceRNA-103571 and IFN-γin the two groups were analyzed;the correlation between hsa-miR-20b and IRF8,hsa-circRNA-103571,IFN-γin patients with pulmonary tuberculosis was analyzed;the factors influencing the occurrence of pulmonary tuberculosis were analyzed by regression analysis;the diagnostic efficiency of serum hsa-miR-20b in pulmonary tuberculosis was analyzed.Results The expression levels of serum hsa-miR-20b,GBP5 mRNA and CUL4A mRNA in pulmonary tuberculosis group were(3.35±0.20),(2.94±0.23)and(0.41±0.15),and those in the control group were(1.01±0.23),(1.02±0.13)and(1.03±0.11).Compared with the control group,the expression levels of serum hsa-miR-20b and GBP5 mRNA in pulmonary tuberculosis group increased,while the expression level of CUL4A mRNA decreased.The difference was statistically significant(P<0.05).The pulmonary tuberculosis group had serum IRF8 of(389.39±46.85)pg/ml and hsa-circRNA-103571 of(22.48±5.06)ng/L,which were higher than(327.62±41.76)pg/ml and(11.36±3.17)ng/L in the control group;the pulmonary tuberculosis group had significantly lower IFN-γof(5.58±1.36)pg/ml than(12.72±4.78)ng/L in the control group;the difference was statistically significant(P<0.05).Pearson correlation analysis showed that serum hsa-miR-20b was positively correlated with IRF8 and hsa-circRNA-103571 levels in patients with pulmonary tuberculosis(r=0.585,0.539;P<0.05),and was negatively correlated with IFN-γlevel(r=-0.485,P<0.05).Logistic regression analysis was performed with the levels of hsa-miR-20b,IRF8,hsa-circRNA-103571,and IFN-γas independent variables,and whether tuberculosis occurred or not as the dependent variable,and the results showed that hsa-miR-20b,IRF8,and hsa-circRNA-103571 were the risk factors for the occurrence of pulmonary tuberculosis(P<0.05),and IFN-γwas a protective factor for the development of pulmonary tuberculosis(P<0.05).The area under the curve(AUC)of serum hsa-miR-20b level for the diagnosis of pulmonary tuberculosis was 0.898,the cut-off value was 1.44,and its sensitivity was 83.82%and specificity was 88.03%.Conclusion There is high expression of hsa-miR-20b in the serum of patients with pulmonary tuberculosis,which is closely related to inflammation-related factors.Hsa-miR-20b may interact with inflammation-related factors to regulate the occurrence and development of pulmonary tuberculosis.