Ding Ziqi, Xiao Xinru, Fan Liang, Mao Zhengdao, Sun Chuang, Li Na, Zhang Qian
Department of Respiratory and Critical Care Medicine, The Affiliated Changzhou Second People's Hospital of Nanjing Medical University, Changzhou, 213164, China.
Changzhou Medical Center, Nanjing Medical University, Changzhou, 213164, China.
Allergy Asthma Clin Immunol. 2024 Mar 23;20(1):23. doi: 10.1186/s13223-024-00890-y.
Circular RNA (circRNA) has the potential to serve as a crucial regulator in the progression of bronchial asthma. The objective of this investigation was to elucidate the functional dynamics of the circ_0070934/miR-199a-5p/Mannoside acetylglucosaminyltransferase 3 (MGAT3) axis in the development of asthma.
Circ_0070934, miR-199a-5p and MGAT3 in peripheral venous blood of 38 asthmatic patients and 43 healthy controls were detected by qRT-PCR, and the expression of MGAT3 protein was examined by ELISA. The GSE148000 dataset was analyzed for differences in MGAT3. The BEAS-2B cells were transfected with circ_0070934 plasmid and small interfering RNA, miR-199a-5p mimics and inhibitors. The apoptosis level was detected by flow cytometry and MGAT3 was detected by qRT-PCR and Western blot. The expression of E-cadherin, N-cadherin, Vimentin was examined by Western blot. Interleukin-4 (IL-4) and IL-13 were used to co-stimulate BEAS-2B cells as an asthmatic airway epithelial cell model. BEAS-2B cells exposed to type 2 cytokines (IL-4 and IL-13) were treated with circ_0070934 plasmid, and the expression of E-cadherin, N-cadherin, and Vimentin was detected by Western blot. The binding relationships were verified using dual-luciferase reporter assay and miRNA pull-down assay.
The expression of circ_0070934 and MGAT3 in peripheral venous blood of asthmatic patients was down-regulated, and the expression of miR-199a-5p was up-regulated. And the expression of MGAT3 was reduced in sputum of asthma patients. Down-regulating the expression of circ_0070934 could promote apoptosis of BEAS-2B cells and increase epithelial-mesenchymal transition (EMT), and this effect can be partially reversed by down-regulating miR-199a-5p. Circ_0070934 could inhibit the process of epithelial mesenchymal transition induced by IL-4 and IL-13 in BEAS-2B cells. In addition, miR-199a-5p could respectively bind to circ_0070934 and MGAT3.
The findings of this study indicate that circ_0070934 may function as a competitive endogenous RNA (ceRNA) of miR-199a-5p, thereby modulating the expression of MGAT3 and impacting the process of EMT in bronchial epithelial cells. These results contribute to the establishment of a theoretical framework for advancing the prevention and treatment strategies for asthma.
环状RNA（circRNA）有可能成为支气管哮喘进展中的关键调节因子。本研究的目的是阐明circ_0070934/miR-199a-5p/甘露糖苷乙酰葡糖胺基转移酶3（MGAT3）轴在哮喘发展中的功能动态。
采用qRT-PCR检测38例哮喘患者和43例健康对照者外周静脉血中circ_0070934、miR-199a-5p和MGAT3的表达，并采用ELISA检测MGAT3蛋白的表达。分析GSE148000数据集以比较MGAT3的差异。用circ_0070934质粒和小干扰RNA、miR-199a-5p模拟物和抑制剂转染BEAS-2B细胞。通过流式细胞术检测细胞凋亡水平，通过qRT-PCR和蛋白质印迹法检测MGAT3。通过蛋白质印迹法检测E-钙黏蛋白、N-钙黏蛋白、波形蛋白的表达。使用白细胞介素-4（IL-4）和IL-13共同刺激BEAS-2B细胞作为哮喘气道上皮细胞模型。用circ_0070934质粒处理暴露于2型细胞因子（IL-4和IL-13）的BEAS-2B细胞，并通过蛋白质印迹法检测E-钙黏蛋白、N-钙黏蛋白和波形蛋白的表达。使用双荧光素酶报告基因测定法和miRNA下拉测定法验证结合关系。
哮喘患者外周静脉血中circ_0070934和MGAT3的表达下调，miR-199a-5p的表达上调。哮喘患者痰液中MGAT3的表达降低。下调circ_0070934的表达可促进BEAS-2B细胞凋亡并增加上皮-间质转化（EMT），下调miR-199a-5p可部分逆转这种作用。Circ_0070934可抑制IL-4和IL-13诱导的BEAS-2B细胞上皮间质转化过程。此外，miR-199a-5p可分别与circ_0070934和MGAT3结合。
本研究结果表明，circ_
Zotero 插件
