Hu Min, Qiu Hui, He Tao, Zhong Minyu
Department of Vascular Surgery, Wuhan Central Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, China.
Department of Oncology, Wuhan NO.1 Hospital, Wuhan 430000, China.
Iran J Public Health. 2022 Nov;51(11):2494-2503. doi: 10.18502/ijph.v51i11.11166.
To explore the effects of miRNA-218-5p on inflammation and extracellular matrix proteins of vascular smooth muscle cell line in abdominal aortic aneurysm (AAA).
miR-218-5p expression was detected with RT-qPCR. The proliferative activity of vascular smooth muscle cells (VSMCs) was detected with CCK-8, the migration was detected by Transwell, and the apoptosis was detected with flow cytometry. The expression levels of inflammatory factors (IL-1β and IL-18) were detected by ELISA. The expression levels of proteins (MMP-9 and Netrin-1) and ADAMTS5 were detected by Western blot. The targeting relationship between miR-218-5p and ADAMTS5 was verified with dual-luciferase reporter assay.
Up-regulating miR-218-5p could significantly inhibit the proliferation and migration of VSMCs and induced the apoptosis (<0.05). Down-regulating miR-218-5p could significantly promote the proliferation and migration of VSMCs and inhibit the apoptosis (<0.05). Up-regulating miR-218-5p could inhibit the expression levels of THP-1 cytoinflammatory factors (IL-8 and IL-1β), MMP-9 and netrin-1. ADAMTS5 was the target gene of miR-218-5p. When there were both overexpression of ADAMTS5 and upregulation of miR-218-5p, the upregulation of miR-218-5p could alleviate the effects of overexpression of ADAMTS5 on the proliferation, migration and apoptosis of VSMCs.
miR-218-5p/ADAMTS-5 molecular axis regulates the proliferation, migration, and apoptosis of VSMCs, as well as the expression of THP-1 cell inflammatory molecules and extracellular matrix molecules.
探讨微小RNA-218-5p(miRNA-218-5p)对腹主动脉瘤(AAA)血管平滑肌细胞系炎症及细胞外基质蛋白的影响。
采用逆转录定量聚合酶链反应(RT-qPCR)检测miR-218-5p表达。用细胞计数试剂盒-8(CCK-8)检测血管平滑肌细胞(VSMCs)的增殖活性,用Transwell检测迁移能力,用流式细胞术检测凋亡情况。采用酶联免疫吸附测定(ELISA)检测炎症因子(白细胞介素-1β和白细胞介素-18)的表达水平。采用蛋白质免疫印迹法检测蛋白(基质金属蛋白酶-9和网蛋白-1)及含血小板反应蛋白基序的解聚素样金属蛋白酶5(ADAMTS5)的表达水平。用双荧光素酶报告基因检测法验证miR-218-5p与ADAMTS5的靶向关系。
上调miR-218-5p可显著抑制VSMCs的增殖和迁移并诱导凋亡(P<0.05)。下调miR-218-5p可显著促进VSMCs的增殖和迁移并抑制凋亡(P<0.05)。上调miR-218-5p可抑制单核细胞系细胞(THP-1)细胞炎症因子(白细胞介素-8和白细胞介素-1β)、基质金属蛋白酶-9和网蛋白-1的表达水平。ADAMTS5是miR-218-5p的靶基因。当ADAMTS5过表达且miR-218-5p上调时,miR-218-5p上调可减轻ADAMTS5过表达对VSMCs增殖、迁移和凋亡的影响。
miR-218-5p/ADAMTS-5分子轴调节VSMCs的增殖、迁移和凋亡,以及THP-1细胞炎症分子和细胞外基质分子的表达。