Yang Lei, Dai Qian, Bao Xiaoming, Li Wang, Liu Jie
Department of Cardiac Surgery, Liaocheng People's Hospital, Liaocheng, 252000 Shandong China.
Department of Geriatric Diseases, Changyi People's Hospital, Changyi, 261300 Shandong China.
Cytotechnology. 2024 Apr;76(2):179-190. doi: 10.1007/s10616-023-00607-w. Epub 2023 Dec 1.
In order to investigate miR-4763-3p and associated genes' roles in myocarditis, AC16 cell line was divided into LPS + miR-4763-3p inhibitor, LPS + NC inhibitor, LPS + miR-4763-3p inhibitor + si-IL10RA and NC groups, and Q-PCR was used to find out whether miR-4763-3p was expressed; Targetscan, Genecards, and MiRDB were used to estimate the miR-4763-3p target; Targetscan was used to display binding sites. Western blot assay was undertaken to detect Bax, Bcl-2, and IL10RA expression. Proliferation and apoptosis were processed using CCK8 and the flow cytometry assay, respectively. Migration and invasion were confirmed utilizing Transwell test. ELISA assay was processed to show the content of IL-6, IL-1ß, IL-10 and TGF-ß in the cell culture supernatant. After being exposed to LPS, cardiomyocyte cells expressed more miR-4763-3p. MiR-4763-3p inhibitor accelerated proliferation, migration and invasion behavior, while it also decreased apoptosis rate in LPS-treated cardiomyocyte cells. MiR-4763-3p inhibitor attenuated the inflammatory response by up-regulating Bax expression and down-regulating Bcl-2 level in LPS-treated cardiomyocyte cells. In cardiomyocyte cells treated with LPS, MiR-4763-3p expression was elevated. si-IL10RA The miR-4763-3p inhibitor restored its effects. MiR-4763-3p accelerates lipopolysaccharide-induced cardiomyocyte apoptosis and inflammatory response by targeting IL10RA, which might be a potential target for myocarditis.
为了研究miR-4763-3p及其相关基因在心肌炎中的作用,将AC16细胞系分为LPS+miR-4763-3p抑制剂组、LPS+NC抑制剂组、LPS+miR-4763-3p抑制剂+si-IL10RA组和NC组,采用Q-PCR检测miR-4763-3p是否表达;使用Targetscan、Genecards和MiRDB评估miR-4763-3p的靶标;用Targetscan显示结合位点。采用蛋白质免疫印迹法检测Bax、Bcl-2和IL10RA的表达。分别使用CCK8和流式细胞术检测细胞增殖和凋亡。利用Transwell实验检测细胞迁移和侵袭能力。采用ELISA法检测细胞培养上清液中IL-6、IL-1β、IL-10和TGF-β的含量。暴露于LPS后,心肌细胞表达更多的miR-4763-3p。miR-4763-3p抑制剂可促进LPS处理的心肌细胞的增殖、迁移和侵袭行为,同时降低其凋亡率。miR-4763-3p抑制剂通过上调LPS处理的心肌细胞中Bax的表达和下调Bcl-2水平来减轻炎症反应。在用LPS处理的心肌细胞中,miR-4763-3p表达升高。si-IL10RA可恢复miR-4763-3p抑制剂的作用。miR-4763-3p通过靶向IL10RA加速脂多糖诱导的心肌细胞凋亡和炎症反应,这可能是心肌炎的一个潜在靶点。