miR-149-5p在白藜芦醇减轻LPS诱导的大鼠心肌细胞损伤中的作用  

Role of microRNA-149-5p in resveratrol-induced reduction of lipopolysaccharide-induced cardiomyocyte injury in rats

作　　者：郝建东 王晓丽[2] 曹婉莹 张宇轩[2] 李青青 徐桂萍[2] Hao Jiandong;Wang Xiaoli;Cao Wanying;Zhang Yuxuan;Li Qingqing;Xu Guiping(Graduate School of Xinjiang Medical University,Urumqi 830000,China;Department of Anesthesiology,People′s Hospital of Xinjiang Uygur Autonomous Region,Xinjiang Clinical Research Center for Anesthesia Management,Urumqi 830000,China)

机构地区：[1]新疆医科大学研究生学院,乌鲁木齐830000 [2]新疆维吾尔自治区人民医院麻醉科、新疆麻醉管理临床医学研究中心,乌鲁木齐830000

出　　处：《中华麻醉学杂志》2024年第7期861-865,共5页Chinese Journal of Anesthesiology

基　　金：新疆维吾尔自治区自然科学基金(2022D01C612);自治区卫生健康青年医学科技人才专项科研项目(WJWY-202325)。

摘　　要：目的评价微小RNA-149-5p(miR-149-5p)在白藜芦醇减轻LPS诱导的大鼠心肌细胞损伤中的作用。方法大鼠心肌细胞系H9C2经培养后,采用随机数字表法分为5组(n=27):对照组(C组)、LPS组、白藜芦醇组(RSV组)、miR149-5p抑制剂阴性对照组(LRN组)和miR149-5p抑制剂组(LRI组)。采用含10μg/ml LPS的培养液孵育细胞24 h制备心肌细胞损伤模型。RSV组经白藜芦醇(终浓度10μmol/L)孵育24 h,随后用含10μg/ml LPS的培养液孵育24 h;LRN组和LRI组分别转染miR149-5p抑制剂阴性对照品和miR149-5p抑制剂,随后操作同RSV组。采用CCK-8法检测细胞活力,流式细胞仪检测细胞凋亡率,微板法检测上清液LDH浓度和细胞还原型谷胱甘肽(GSH)含量,TBA反应法检测MDA含量,WST-1法检测SOD活性,DCFH-DA荧光探针检测ROS水平,ELISA法检测上清液TNF-α和IL-6浓度,RT-qPCR法检测miR-149-5p表达。结果与C组比较,LPS组miR-149-5p表达下调,细胞活力降低,上清液LDH、TNF-α和IL-6浓度、细胞凋亡率、ROS水平和MDA含量升高,GSH含量和SOD活性降低(P<0.05)。与LPS组比较,RSV组miR-149-5p表达上调,细胞活力升高,上清液LDH、TNF-α和IL-6浓度、细胞凋亡率、ROS水平和MDA含量降低,GSH含量和SOD活性升高(P<0.05)。与RSV组或LRN组比较,LRI组细胞miR-149-5p表达下调,细胞活力降低,上清液LDH、TNF-α和IL-6浓度、细胞凋亡率、ROS水平和MDA含量升高,GSH含量和SOD活性降低(P<0.05)。结论白藜芦醇减轻LPS诱导大鼠心肌细胞损伤的机制与上调miR-149-5p表达,抑制细胞凋亡、氧化应激和炎症反应有关。Objective To evaluate the role of microRNA-149-5p(miR-149-5p)in resveratrol-induced reduction of lipopolysaccharide(LPS)-induced cardiomyocyte injury in rats.Methods Rat cardiomyocyte cell line H9C2 was cultured and then divided into 5 groups(n=27 each)using a random number method:control group(C group),LPS group,resveratrol group(RSV group),miR149-5p inhibitor negative control group(LRN group),and miR149-5p inhibitor group(LRI group).A cardiomyocyte injury model was prepared by incubating cells with culture medium containing 10μg/ml LPS for 24 h.RSV group was incubated with resveratrol(final concentration of 10μmol/L)for 24 h,followed by incubation with culture medium containing 10μg/ml LPS for another 24 h.LRN group and LRI group were transfected with miR149-5p inhibitor negative control and miR149-5p inhibitor,respectively,and then the other treatments were similar to those previously described in RSV group.The cell viability was measured by CCK-8 assay,the apoptosis rate by flow cytometry,the concentration of lactate dehydrogenase(LDH)and content of glutathione(GSH)in the supernatant by microplate method,the content of malondialdehyde(MDA)by TBA reaction method,the activity of superoxide dismutase(SOD)by WST-1 method,the level of reactive oxygen species(ROS)by DCFH-DA fluorescent probe,the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the supernatant by enzyme-linked immunosorbent assay,and the expression of miR-149-5p by quantitative real-time polymerase chain reaction.Results Compared with C group,the expression of miR-149-5p was significantly down-regulated,the cell viability was decreased,the concentrations of LDH,TNF-αand IL-6 in supernatant,apoptosis rate,ROS level and MDA content were increased,and the GSH content and SOD activity were decreased in LPS group(P<0.05).Compared with LPS group,the expression of miR-149-5p was significantly up-regulated,the cell viability was increased,the concentrations of LDH,TNF-αand IL-6 in supernatant,apoptosis rate,ROS level and M

关 键 词：白藜芦醇 微RNA 肌细胞 心脏 脂多糖类 细胞凋亡 炎症 氧化应激 

分 类 号：R285.5[医药卫生—中药学]