Fan Linxia, Wang Xiaojun, Fan Linlan, Chen Qizhang, Zhang Hong, Pan Hui, Xu Aixia, Wang Hongjuan, Yu Yang
a Respiratory Department in the Cadre Ward , Gansu Provincial Hospital , Lanzhou , China.
b Respiratory Department , Gansu Provincial Hospital , Lanzhou , China.
Exp Lung Res. 2016 Oct-Dec;42(8-10):417-424. doi: 10.1080/01902148.2016.1256452. Epub 2016 Nov 30.
To delineate the underlying mechanism of microRNA-145 modulate the balance of Th1/Th2 via targeting RUNX3 in asthma patients.
Peripheral blood samples were collected from asthma patients and healthy controls. CD4 T cells were isolated and cultured. Using quantitative PCR detect, the level of microRNA-145 and RUNX3 mRNA level in the CD4 T cells from asthma patients and healthy controls, meanwhile, western blot was used to detect the RUNX3 protein level. Th1 or Th2 related cytokines were measured by enzyme-linked immunosorbent assay. Dual-Luciferase Reporter Assay was performed to confirm the correlation between microRNA-145 and RUNX3. MicroRNA-145 mimic or inhibitor was transfected in the CD4 T cells and the changes of RUNX3 level, Th1 or Th2 related cytokines and the percentage of Th1 and Th2 were observed after transfection.
MicroRNA-145 level of CD4 T cells was higher with a lower RUNX3 expression in asthma patients. There is negative correlation between microRNA-145 and RUNX3. Th2 hyperactivity and Th1 deficiency was detected in the CD4 T cells of asthma patients. Dual-Luciferase Reporter Assay has shown that RUNX3 is a target of microRNA. Up-regulation or down-regulation of miR-145 level caused RUNX3 expression changes in CD4 T cells and influence the related cytokines. Inhibition of microRNA-145 may reverse the imbalance of Th1/Th2 in asthma patients.
MicroRNA-145 could regulate the balance of Th1/Th2 through targeting the RUNX3 in asthma patients. MicroRNA-145 and RUNX3 may be used as biomarkers or targets in the diagnosis or therapy of asthma.
探讨微小RNA-145通过靶向RUNX3调控哮喘患者Th1/Th2平衡的潜在机制。
采集哮喘患者和健康对照者的外周血样本。分离并培养CD4 T细胞。采用定量PCR检测哮喘患者和健康对照者CD4 T细胞中微小RNA-145水平及RUNX3 mRNA水平，同时用蛋白质免疫印迹法检测RUNX3蛋白水平。采用酶联免疫吸附测定法检测Th1或Th2相关细胞因子。进行双荧光素酶报告基因检测以证实微小RNA-145与RUNX3之间的相关性。将微小RNA-145模拟物或抑制剂转染至CD4 T细胞中，观察转染后RUNX3水平、Th1或Th2相关细胞因子以及Th1和Th2百分比的变化。
哮喘患者CD4 T细胞中微小RNA-145水平较高，而RUNX3表达较低。微小RNA-145与RUNX3呈负相关。在哮喘患者的CD4 T细胞中检测到Th2功能亢进和Th1缺陷。双荧光素酶报告基因检测表明RUNX3是微小RNA的靶标。上调或下调miR-145水平会导致CD4 T细胞中RUNX3表达改变，并影响相关细胞因子。抑制微小RNA-145可能会逆转哮喘患者Th1/Th2的失衡。
在哮喘患者中，微小RNA-145可通过靶向RUNX3调节Th1/Th2平衡。微小RNA-145和RUNX3可作为哮喘诊断或治疗的生物标志物或靶点。
