MiR-205-3p靶向PRMT5改善P.g-LPS所致HUVECs炎症反应和凋亡  

MiR-205-3p targets PRMT5 to improve inflammatory response and apoptosis of HUVECs induced by P.g-LPS

作　　者：吴金生 李佳 李维善[1] WU Jin-sheng;LI Jia;LI Wei-shan(Department of Periodontal Mucosal Diseases,Affiliated Stomatological Hospital of Jiamusi University,Jiamusi 154000,China)

机构地区：[1]佳木斯大学附属口腔医院牙周黏膜病科,黑龙江佳木斯154000

出　　处：《牙体牙髓牙周病学杂志》2024年第3期132-141,共10页Chinese Journal of Conservative Dentistry

基　　金：黑龙江省教育厅基本科研业务费基础研究项目(2019-KYYWF-1360)。

摘　　要：目的:研究miR-205-3p对P.g-LPS所致HUVECs炎症反应和凋亡的影响及调控机制。方法:利用P.g-LPS刺激HUVECs构建细胞损伤模型,实验分为Con组、LPS组、LPS+miR-NC组、LPS+miR-205组、LPS+si-NC组、LPS+si-PRMT5组、LPS+miR-205+pc-NC组和LPS+miR-205+pc-PRMT5组,RT-qPCR和Western blot检测miR-205-3p、PRMT5和凋亡蛋白表达;ELISA检测炎症因子表达;Tunel染色和流式细胞术检测HUVECs凋亡;双荧光素酶报告基因实验和Western blot验证miR-205-3p和PRMT5的靶向关系。结果:P.g-LPS刺激HUVECs后,炎症因子表达和凋亡率显著升高,miR-205-3p下调,PRMT5上调(P <0.05)。miR-205-3p可以抑制PRMT5表达,且过表达miR-205-3p和PRMT5低表达均可以降低炎症因子表达和降低细胞凋亡率(P <0.05)。双荧光素酶报告基因实验和Western blot结果表明miR-205-3p可靶向抑制PRMT5的表达(P <0.05)。且过表达PRMT5可明显逆转过表达miR-205-3p对炎症反应和凋亡的抑制作用(P <0.05)。结论:P.g-LPS促进HUVECs炎症反应和凋亡;miR-205-3p靶向负调控PRMT5表达来改善P.g-LPS诱导的HUVECs炎症反应和凋亡。AIM:To investigate the effect of miR-205-3p on the inflammatory response and apoptosis of HU-VECs induced by P.g-LPS and its regulatory mechanism,METHODS:HUVECs were stimulated with P.g-LPS to estab-lish a cell injury model.The experiment was divided into Con group,LPS group,LPS+miR-NC group,LPS+miR-205 group,LPS+si-NC group,LPS+si-PRMT5 group,LPS+miR-205+pc-NC group,and LPS+miR-205+pc-PRMT5 group.RT-qPCR and Western blot were used to detect the expression of miR-205-3p,PRMT5,and apoptosis proteins.ELISA was used to detect the expression of inflammatory factors.Tunel staining and flow cytometry were used to detect HUVECs apoptosis.The dual luciferase reporter gene assay and Western blot were used to verify the targeted relationship between miR-205-3p and PRMT5.RESULTS:After P.g-LPS stimulation of HUVECs,the expression of inflammatory factors and apoptosis rate significantly increased,while miR-205-3p was down-regulated and PRMT5 was up-regulated(P<0.05).miR-205-3p can inhibit PRMT5 expression,and overexpression of miR-205-3p and low expression of PRMT5 can reduce inflammatory factor expression and apoptosis rate(P<0.05).The results of the dual luciferase reporter gene assay and West-erm blot showed that miR-205-3p can target and inhibit the expression of PRMT5(P<0.05).Overexpression of PRMT5 can significantly reverse the inhibitory effect of overexpression of miR-205-3p on inflammatory response and apoptosis(P<0.05).CONCLUSION:P.g-LPS promotes HUVECs inflammatory response and apoptosis;miR-205-3p targets and negatively regulates PRMT5 expression to improve P.g-LPS-induced HUVECs inflammatory response and apoptosis.

关 键 词：MiR-205-3p 蛋白质精氨酸甲基转移酶5 炎症反应 凋亡率 牙龈卟啉单胞菌脂多糖 人脐静内皮细胞 

分 类 号：R781.4[医药卫生—口腔医学]