黄芩苷对脂多糖诱导心肌细胞炎症反应和焦亡的抑制作用及其机制  

Inhibitory effects of baicalin on LPS-induced inflammation and pyroptosis of cardiomyocytes and their mechanism

作　　者：靳宜静 李堪董 詹智晖 王勇 JIN Yijing;LI Kandong;ZHAN Zhihui;WANG Yong(Department of Cardiovascular Medicine,Hainan Western Central Hospital,Danzhou 571000,China;不详)

机构地区：[1]海南西部中心医院心血管内科,海南儋州571000 [2]海南西部中心医院神经外科

出　　处：《山东医药》2024年第5期12-16,共5页Shandong Medical Journal

基　　金：海南省卫生健康行业科研项目(21A200325)。

摘　　要：目的观察黄芩苷对脂多糖(LPS)诱导心肌细胞焦亡、炎症反应的抑制作用并分析其机制。方法体外培养大鼠心肌细胞H9C2,取对数生长期细胞分为对照组、LPS组、黄芩苷组,对照组不做干预,LPS组以LPS刺激H9C2细胞构建体外细胞损伤模型,黄芩苷组在LPS诱导后分别加入10、20、40、80μmol/L黄芩苷,CCK-8法检测各组细胞活力,取细胞活力最高的黄芩苷浓度作为最佳作用浓度进行后续实验。而后取对数期H9C2细胞分为对照组、LPS组、黄芩苷组、抑制剂组、黄芩苷+抑制剂组及黄芩苷+激活剂组,除不做干预的对照组外均给予LPS刺激构建细胞损伤模型;黄芩苷组在此基础上给予黄芩苷处理,抑制剂组给予磷脂酰肌醇3激酶(PI3K)/丝苏氨酸蛋白激酶(AKT)通路抑制剂LY294002处理,黄芩苷+抑制剂组给予黄芩苷及LY294002处理,黄芩苷+激活剂组给予黄芩苷及PI3K/AKT通路激活剂胰岛素样生长因子1(IGF-1)处理。采用ELISA法检测细胞上清液炎症因子白细胞介素(IL)-1β、IL-6,Western blotting法检测细胞焦亡相关蛋白焦孔素D(GSDMD)、核苷酸寡聚化结构域样受体蛋白3(NLRP3)、半胱氨酸蛋白酶1(Caspase-1)及PI3K/AKT通路相关蛋白PI3K、AKT、p-PI3K、p-AKT相对表达量。结果与对照组比较,LPS组细胞上清液炎症因子IL-6、IL-1β水平升高;与LPS组比较,黄芩苷组、抑制剂组IL-6、IL-1β水平降低;与黄芩苷组比较,黄芩苷+抑制剂组IL-6、IL-1β水平降低,黄芩苷+激活剂组IL-6、IL-1β水平升高(P均<0.05)。与对照组比较,LPS组细胞焦亡相关蛋白GSDMD、NLRP3、Caspase-1表达升高;与LPS组比较,黄芩苷组、抑制剂组GSDMD、NLRP3、Caspase-1蛋白表达水平降低;与黄芩苷组比较,黄芩苷+抑制剂组GSDMD、NLRP3、Caspase-1蛋白表达降低,黄芩苷+激活剂组GSDMD、NLRP3、Caspase-1蛋白表达升高(P均<0.05)。与对照组比较,LPS组PI3K/AKT通路相关蛋白p-PI3K、p-AKT表达升高;与LPS组比较,黄芩�Objective To observe the inhibitory effects of baicalin on lipopolysaccharide(LPS)-induced pyroptosis and inflammation of cardiomyocytes and to analyze their mechanism.Methods The rat cardiomyocytes H9C2 were cultivated in vitro and we divided cells in the logarithmic growth phase into the control group,LPS group,and baicalin group.The control group did not receive intervention.In the LPS group,H9C2 cells were stimulated by LPS to construct the cell injury models in vitro.In the baicalin group,cells were induced by LPS and then were added with 10,20,40,and 80μmol/L baicalin,respectively.The cell viability in each group was detected by CCK-8.The concentration of baicalin with the highest cell viability was selected as the optimal concentration for subsequent experiments.Then,H9C2 cells in the logarithmic phase were divided into the control group,LPS group,baicalin group,inhibitor group,baicalin+inhibitor group,and baicalin+activator group.Except the control group without intervention,LPS stimulation was conducted to construct the cell injury models in the other groups;on this basis,the baicalin group was treated with baicalin,while the inhibitor group was treated with phosphatidylinositol 3-kinase(PI3K)/serine protein kinase(AKT)pathway inhibitor LY294002,the baicalin+inhibitor group was treated with baicalin and LY294002,and the baicalin+activator group was treated with baicalin and PI3K/AKT pathway activator insulin-like growth factor 1(IGF-1).The inflammatory cytokines interleukin(IL)-1βand IL-6 in the cell supernatant were detected by ELISA.Western blotting was used to detect the relative expression levels of pyroptosis-related proteins pyroporin D(GSDMD),nucleotide oligomeric domain-like receptor protein 3(NLRP3),Caspase-1,and PI3K/AKT pathway-related proteins PI3K,AKT,phosphorylation(p-)PI3K,and p-AKT.Results Compared with the control group,the expression levels of inflammatory cytokines IL-6,IL-1β,pyroptosis-related proteins GSDMD,NLRP3,Caspase-1,p-PI3K and p-AKT increased in the LPS group.Compared with

关 键 词：黄芩苷 心肌损伤 炎症反应 细胞焦亡 磷脂酰肌醇3激酶 丝苏氨酸蛋白激酶 

分 类 号：R734.2[医药卫生—肿瘤]