结核病患者中靶向调控维生素D受体的microRNA的初步筛选

目的筛选与结核病相关，且靶向调控维生素D受体(vitamin D receptor, VDR)的microRNA(miRNA)，为深入研究维生素D(vitamin D, VitD)辅助抗结核病机制及其临床应用提供一定的参考依据。 方法选择5例明确诊断肺结核病患者和5例非结核病的志愿者作为研究组和对照组。分别收集临床基线资料以及采集两组外周血单个核细胞(peripheral blood mononuclear cell, PBMC)作为临床样本，提取总RNA后，采用高通量测序技术(high-throughput sequencing)，检测两组间的miRNA差异表达情况。 结果5例肺结核病患者和5例非结核病志愿者临床基线资料经统计学分析得出两组间23项指标的同质性和异质性，其中性别、年龄、吸烟饮酒习惯等两组间无统计学差异；体重指数(BMI)、总胆固醇(TC)、血红蛋白(Hb)白蛋白(ALB)、球蛋白(GLB)及总蛋白(TP)研究组低于对照组，有统计学差异(P<0.05)。高通量测序从研究组和对照组5对样本中共筛选出1 560条miRNAs，得到miRNA在两组中差异表达谱。分层聚类和火山图进一步综合分析发现研究组中最有差异的23条miRNAs(P<0.05)，其中15条miRNAs在研究组中表达下(∣LOG2FC∣>1)，8条miRNAs表达上调(FC>1)。综合高通量测序结果和生物信息学预测，从差异表达谱中筛选出2条有统计学意义且靶向VDR的miRNAs：hsa-miR-326(P<0.05、FC＝1.37)、hsa-miR-654-5p(P<0.05、∣LOG2FC∣＝1.06)。 结论通过临床样本高通量测序分析及生物信息学预测，发现了结核病中的既差异表达又靶向VDR的两条miRNAs，这两条miRNAs可能是肺结核病潜在的诊断标志及新的治疗靶点，为结核病的防治提供了新的策略和方向。

A study of microRNA Prescreening that targeted and regulated Vitamin D rceptor in tuberculosis patients

ObjectiveMicroRNA with significantly differential expression and targeted regulation of Vitamin D receptor(VDR) in Tuberculosis were screened by High-throughput sequencing and bioinformatics methods, whichprovided the further study of Vitamin D anti-tuberculosis mechanism and clinical application. MethodAccording to the inclusion and exclusion criteria, 5 patients with definite diagnosis of tuberculosis and 5 non-tuberculosis volunteers were selected as the study group and control group. Peripheral blood mononuclear cells(PBMC) of the two groups were collected as clinical samples. After total RNA was extracted, high-throughput sequencing technology was used to detect the differential expression of miRNA between the two groups. ResultThe clinical baseline data of 5 tuberculosis patients and 5 non-tuberculosis volunteers were collected, and the homogeneity and heterogeneity of 23 items between the two groups were obtained by statistical analysis. It was found that there were no significatly statistical differences in gender, age, smoking and drinking habits between the two groups. Body mass index (BMI), total cholesterol (TC), hemoglobin (Hb) albumin (ALB), globulin (GLB) and total protein (TP) in the study group were lower than those in the control group with significatly statistical differences(P<0.05). A total of 1560 miRNAs were screened out by miRNA sequencing. Through stratified cluster and comprehensive volcanic map analysis, 23 miRNAs with the most significant (P<0.05) differences were found in the case group, among which 15 were down-regulated(∣LOG2FC∣>1) and 8 were up-regulated(FC>1). In short, Combined with bioinformatics predictions, two miRNAs targeted VDR that differentially expressed in tuberculosis patients were screened out: hsa-miR-326(P<0.05、FC=1.37)、hsa-miR-654-5p(P<0.05、∣LOG2FC∣=1.06). ConclusionThese two differentially expressed miRNAs mentioned abovemay be potential diagnostic markers and new therapeutic targets for tuberculosis.This found provides a new strategy and feasible direction for the prevention and treatment of Tuberculosis.