NLRP3炎性小体在脂多糖诱导的高糖心肌细胞缺氧复氧损伤中的作用及机制  

Role and Mechanism of NLRP3 Inflammasome in Lipopolysaccharide-induced Hypoxia and Reoxygenation Injury of Hyperglycemic Cardiomyocytes

作　　者：李璐[1] 邱珍[1] 张艺[1] 田浩 夏中元[1] LI Lu;QIU Zhen;ZHANG Yi(Department of Anesthesiology,Renmin Hospital of Wuhan University,Hubei 430000,China)

机构地区：[1]武汉大学人民医院麻醉科,430000

出　　处：《医学研究杂志》2022年第9期44-48,共5页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81970722);武汉大学自主科研项目(2042021kf0113)。

摘　　要：目的 探究NLRP3炎性小体在脂多糖(lipopolysaccharide, LPS)诱导的心肌细胞高糖缺氧/复氧(hypoxia/reoxygenation, H/R)损伤中的作用及机制。方法 取正常对数期生长的H9C2心肌细胞,随机分为4组,即高糖(H)组(n=3)、高糖+缺氧/复氧(H+H/R)组(n=3)、高糖+LPS+H/R(H+LPS+H/R)组(n=3)和BAY11-7082干预高糖+LPS+H/R(H+LPS+BAY+H/R)组(n=3)。采用CCK-8法检测细胞活力,ROS检测试剂盒测定线粒体氧化应激水平,RT-PCR检测细胞NLRP3、ASC、caspase-1和IL-1β的mRNA表达水平,Western blot法检测细胞NLRP3、ASC、caspase-1和IL-1β的蛋白表达水平。结果 与高糖组比较,H+H/R组细胞活性降低、LDH水平升高、线粒体ROS产生增加,NLRP3、ASC、caspase-1和IL-1β的mRNA表达上调,NF-κB、NLRP3、ASC、caspase-1和IL-1β蛋白水平增高(P<0.05);LPS处理后,H/R的高糖心肌细胞损伤进一步加重(P<0.05),而加用BAY11-7082后,LPS加重的损伤得以改善(P<0.05)。结论 BAY11-7082可以抑制NLRP3炎性小体激活,从而减轻LPS介导的H9C2细胞高糖H/R损伤。Objective To investigate the role and mechanism of NLRP3 inflammasome in lipopolysaccharide-induced hyperglycemic hypoxia/reoxygenation injury of cardiomyocytes. Methods H9 C2 cardiomyocytes grown at normal logarithmic phase were randomly divided into four groups: high glucose group(H group, n=3), high glucose + hypoxia/reoxygenation group(H+H/R group, n=3), High glucose +LPS+H/R group(H+LPS+H/R group, n=3) and BAY11-7082 intervention high glucose +LPS+H/R group(H+LPS+BAY+H/R group, n=3). Cell viability and mitochondrial oxidative stress(ROS) levels were detected by kit. mRNA expression levels of NLRP3, ASC, caspase-1 and IL-1β were detected by RT-PCR, and protein expression levels of NLRP3, ASC, caspase-1 and IL-1β were detected by Western blot. Results Compared with high glucose group, the cell activity of H+H/R group decreased, LDH level and mitochondrial ROS production increased, mRNA expressions of NLRP3, ASC, caspase-1 and IL-1β up-regulated, and protein levels of NF-κB, NLRP3, ASC, caspase-1 and IL-1β increased(P<0.05). After LPS treatment, the damage of H/R hyperglycemic cardiomyocytes was further aggravated(P<0.05), while the addition of Bay11-7082 improved the damage(P<0.05). Conclusion BAY11-7082 inhibited NLRP3 inflammasome activation and thus alleviated LPS-mediated high glucose H/R injury in H9 C2 cells.

关 键 词：NLRP3炎性小体 脂多糖 缺氧/复氧损伤 高糖 心肌细胞 

分 类 号：R541[医药卫生—心血管疾病]