Zeng Shaolin, Cui Juan, Zhang Yunting, Zheng Zhishui
Department of Respiratory and Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
Department of Critical Care and Intensive Care Medicine, Xiangzhou District People's Hospital, Xiangyang, 441100, Hubei Province, China.
Inflammation. 2022 Aug;45(4):1548-1558. doi: 10.1007/s10753-022-01640-1. Epub 2022 Mar 18.
The dysfunction of airway smooth muscle cells (ASMCs) is one of the key factors in the pathogenesis of asthma. How miR-98-5p works in asthma has not been completely elucidated. This work focused on how miR-98-5p functions in the proliferation and migration of human ASMCs treated with interleukin-13 (IL-13). MiR-98-5p expression in plasma of asthmatic patients and IL-13-stimulated ASMCs was probed by quantitative real-time polymerase chain reaction (qRT-PCR). RAS-relevant C3 botulinum toxin substrate 1 (RAC1) protein expression in ASMCs was assessed by Western blot. The growth of ASMCs was measured by cell counting kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) assay. The migration of ASMCs was examined by Transwell assay. Besides, the apoptosis of ASMCs was analyzed by flow cytometry. The targeting relationship between miR-98-5p and RAC1 3'-UTR was verified by a dual-luciferase reporter gene assay. MiR-98-5p expression was reduced in patients' plasma and IL-13-stimulated ASMCs, and RAC1 expression was upregulated in ASMCs treated with IL-13. MiR-98-5p overexpression inhibited IL-13-induced proliferation and migration of ASMCs while promoting the apoptosis. The opposite result was observed after inhibiting miR-98-5p expression. Besides, RAC1 was identified as a direct downstream target of miR-98-5p in ASMCs. The restoration of RAC1 expression counteracted the impacts of miR-98-5p overexpression on IL-13-stimulated proliferation, migration, and apoptosis of ASMCs. MiR-98-5p inhibits IL-13-induced proliferation and migration and accelerates the apoptosis of ASMCs by downregulating RAC1 expression.
气道平滑肌细胞（ASMCs）功能障碍是哮喘发病机制的关键因素之一。miR-98-5p在哮喘中的作用机制尚未完全阐明。本研究聚焦于miR-98-5p在白细胞介素-13（IL-13）处理的人ASMCs增殖和迁移中的作用。通过定量实时聚合酶链反应（qRT-PCR）检测哮喘患者血浆和IL-13刺激的ASMCs中miR-98-5p的表达。通过蛋白质免疫印迹法评估ASMCs中RAS相关的C3肉毒杆菌毒素底物1（RAC1）蛋白的表达。采用细胞计数试剂盒-8（CCK-8）法和5-乙炔基-2'-脱氧尿苷（EdU）法检测ASMCs的生长情况。采用Transwell法检测ASMCs的迁移情况。此外，通过流式细胞术分析ASMCs的凋亡情况。通过双荧光素酶报告基因检测验证miR-98-5p与RAC1 3'-UTR之间的靶向关系。哮喘患者血浆和IL-13刺激的ASMCs中miR-98-5p表达降低，而IL-13处理的ASMCs中RAC1表达上调。miR-98-5p过表达抑制IL-13诱导的ASMCs增殖和迁移，同时促进其凋亡。抑制miR-98-5p表达后观察到相反的结果。此外，RAC1被确定为ASMCs中miR-98-5p的直接下游靶点。RAC1表达的恢复抵消了miR-98-5p过表达对IL-13刺激的ASMCs增殖、迁移和凋亡的影响。miR-98-5p通过下调RAC1表达抑制IL-13诱导的ASMCs增殖和迁移，并加速其凋亡。
