摘要
目的探讨微小RNA-194-5p(miR-194-5p)与泛素连接酶cullin4A(CUL4A)在肺结核病人中的表达及其意义。方法选取2016年12月至2018年12月中国平煤神马集团职业病防治院结核病病人为研究对象,确诊为肺结核病人50例作为肺结核组,潜伏结核感染病人50例作为潜伏结核感染组,同期该院体检健康者50例作为对照组。实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测miR-194-5p、CUL4A mRNA、鸟苷酸结合蛋白5(GBP5)mRNA的表达;酶联免疫吸附测定(ELISA)检测可溶性Tim-3(sTim-3)、可溶性Gal-9(sGal-9)水平;采用Pearson相关性分析检验肺结核病人miR-194-5p、CUL4A表达量与GBP5、sTim-3、sGal-9的相关性。结核分枝杆菌(Mtb)感染人巨噬细胞系U937,ELISA检测白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)水平。蛋白质印迹法(Western blotting)检测Wnt/β-连环素(Wnt/β-catenin)信号通路相关蛋白T细胞因子(TCF)、糖原合成酶激酶-3(GSK-3β)、β-catenin表达量。结果与对照组相比,潜伏结核感染组与肺结核组病人血清miR-194-5p、GBP5 mRNA的表达水平升高(P<0.05),其中miR-194-5p:(1.01±0.23)比(2.32±0.53)比(3.35±0.20),sTim-3、sGal-9水平升高(P<0.05),CUL4A mRNA的表达水平显著降低[(1.03±0.11)比(0.73±0.12)比(0.41±0.15),P<0.05],潜伏结核感染组与肺结核组各指标间相比差异有统计学意义(P<0.05);miR-194-5p与CUL4A呈负相关(r=-0.421,P<0.001),miR-194-5p与GBP5、sTim-3、sGal-9呈正相关(P<0.05);Mtb感染细胞后,IL-6、TNF-α的表达水平升高(P<0.05),TCF、GSK-3β、β-catenin表达量升高(P<0.05)。结论miR-194-5p在肺结核病人中表达上调,CUL4A表达下调,二者可能参与肺结核发生及发展过程。
Objective To investigate the expression and significance of miR-194-5p and CUL4A in patients with tuberculosis.Methods Patients with tuberculosis from the China Pingmei Shenma Group Occupational Disease Prevention and Treatment Hospital from December 2016 to December 2018 were selected as the research object,and the control group(50 cases),latent combined infec⁃tion group(50 cases),and pulmonary tuberculosis group(50 cases)were set in the experiment.qRT-PCR was used to detect the expres⁃sion of miR-194-5p,CUL4A mRNA and GBP5 mRNA.The levels of sTim-3 and sGal-9 were detected by ELISA.Correlation of miR-194-5p and CUL4A expression with GBP5,sTim-3,and sGal-9 in patients with tuberculosis was analyzed by Pearson method.Mtb in⁃fected human macrophage cell line U937 and detected the levels of IL-6 and TNF-αby ELISA.Western blotting was used to detect the expression of TCF,GSK-3βandβ-catenin.Results Compared with the control group,the expression levels of serum miR-194-5p and GBP5 mRNA in the latent tuberculosis infection group and the pulmonary tuberculosis group were increased(P<0.05),and miR-194-5p:(1.01±0.23)vs.(2.32±0.53)vs.(3.35±0.20),the levels of sTim-3 and sGal-9 were significantly increased(P<0.05),and the expres⁃sion level of CUL4A mRNA was decreased[(1.03±0.11)vs.(0.73±0.12)vs.(0.41±0.15),P<0.05],there was a statistically significant difference between the indicators of the latent tuberculosis infection group and the pulmonary tuberculosis group(P<0.05).miR-194-5p was negatively correlated with CUL4A(r=-0.421,P<0.001),miR-194-5p was positively correlated with GBP5,sTim-3,and sGal-9(P<0.05).After Mtb infected cells,the expression levels of IL-6 and TNF-αwere increased(P<0.05),and the expression levels of TCF,GSK-3βandβ-catenin were increased(P<0.05).Conclusion miR-194-5p was up-regulated in tuberculosis patients and CUL4A was down-regulated.Both may be involved in the occurrence and development of tuberculosis.
作者
闫书娜
马腾
李国锋
李金洲
曾俊风
YAN Shuna;MA Teng;LI Guofeng;LI Jinzhou;ZENG Junfeng(Five Districts of Internal Medicine,Shen-ma Group Occupational Disease Prevention and Control Hospital,Pingdingshan,Henan 467099,China;Lung lavage center,Shen-ma Group Occupational Disease Prevention and Control Hospital,Pingdingshan,Henan 467099,China;Department of Internal Medicine,Shen-ma Group Occupational Disease Prevention and Control Hospital,Pingdingshan,Henan 467099,China;Department of Respiration,General Hospital of China Pingmei Shenma Medical Group,Pingdingshan,Henan 467000,China;Department of Liver Diseases,Central Hospital of Nanyang City,Nanyang,Henan 473000,China)
出处
《安徽医药》
CAS
2021年第3期489-493,共5页
Anhui Medical and Pharmaceutical Journal
