Dou Liyan, Wang Wenyu, Wang Junwei, Zhang Xiaofei, Hu Xiaoman, Zheng Weili, Han Kaiyu, Wang Guangyou
Department of Cardiology, The Fourth Affiliated Hospital of Harbin Medical University, Heilongjiang, China.
Department of Respiratory and Critical Medicine, The Second Affiliated Hospital of Harbin Medical University, Heilongjiang, China.
Allergy Asthma Clin Immunol. 2022 Aug 4;18(1):66. doi: 10.1186/s13223-022-00704-z.
Several miRNAs are now known to have clear connections to the pathogenesis of asthma. The present study focused on the potential role of miR-3934 during asthma development.
miR-3934 was detected as a down-regulated miRNA in basophils by sequencing analysis. Next, the expression levels of miR-3934 in peripheral blood mononuclear cells of 50 asthma patients and 50 healthy volunteers were examined by RT-qPCR methods. The basophils were then treated with AGEs and transfected with miR-3934 mimics. The apoptosis levels were examined by flow cytometry assay; and the expression levels of cytokines were detected using the ELISA kits. Finally, the Western blot was performed to examined the expression of key molecules in the TGF-β/Smad signaling pathway.
miR-3934 was down-regulated in the basophils of asthmatic patients. The expression of the pro-inflammatory cytokines IL-6, IL-8 and IL-33 was enhanced in basophils from asthmatic patients, and this effect was partially reversed by transfection of miR-3934 mimics. Furthermore, receiver operating characteristics analysis showed that miR-3934 levels can be used to distinguish asthma patients from healthy individuals. miR-3934 partially inhibited advanced glycation end products-induced increases in basophil apoptosis by suppressing expression of RAGE.
Our results indicate that miR-3934 acts to mitigate the pathogenesis of asthma by targeting RAGE and suppressing TGF-β/Smad signaling.
现已发现几种微小RNA(miRNA)与哮喘的发病机制有明确关联。本研究聚焦于miR - 3934在哮喘发展过程中的潜在作用。
通过测序分析检测到miR - 3934是嗜碱性粒细胞中下调的miRNA。接下来,采用逆转录定量聚合酶链反应(RT - qPCR)方法检测50例哮喘患者和50名健康志愿者外周血单个核细胞中miR - 3934的表达水平。然后用晚期糖基化终产物(AGEs)处理嗜碱性粒细胞并转染miR - 3934模拟物。通过流式细胞术检测凋亡水平;使用酶联免疫吸附测定(ELISA)试剂盒检测细胞因子的表达水平。最后,进行蛋白质免疫印迹法检测转化生长因子-β(TGF - β)/Smad信号通路中关键分子的表达。
哮喘患者嗜碱性粒细胞中miR - 3934表达下调。哮喘患者嗜碱性粒细胞中促炎细胞因子白细胞介素-6(IL - 6)、白细胞介素-8(IL - 8)和白细胞介素-33(IL - 33)的表达增强,转染miR - 3934模拟物可部分逆转这种作用。此外,受试者工作特征分析表明,miR - 3934水平可用于区分哮喘患者和健康个体。miR - 3934通过抑制晚期糖基化终产物诱导的嗜碱性粒细胞凋亡增加,其机制是通过抑制晚期糖基化终产物受体(RAGE)的表达。
我们的结果表明,miR - 3934通过靶向RAGE并抑制TGF - β/Smad信号通路来减轻哮喘的发病机制。