棕榈油酸在缺氧/复氧心肌细胞焦亡中的作用及机制研究  

The role and mechanism of palmitoleic acid in the pyroptosis of cardiomyocytes after hypoxia/reoxygenation-induced injury

作　　者：李云权 段宗刚 鲍海龙 王清腾 安洪鑫 夏侯鸾达 王旭 蒋孟庭 周海燕 罗振华[3] 李伟 Li Yunquan;Duan Zonggang;Bao Hailong;Wang Qingteng;An Hongxin;Xiahou Luanda;Wang Xu;Jiang Mengting;Zhou Haiyan;Luo Zhenhua;Li Wei(Dept of Cardiovascular Medicine,The Affiliated Hospital of Guizhou Medical University,Guiyang 550004;School of Clinical Medicine,Guizhou Medical University,Guiyang 550004;Dept of Central Lab,Guizhou Provincial People′s Hospital,Guiyang 550002)

机构地区：[1]贵州医科大学附属医院心内科,贵阳550004 [2]贵州医科大学临床医学院,贵阳550004 [3]贵州省人民医院中心实验室,贵阳550002

出　　处：《安徽医科大学学报》2024年第6期1006-1012,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81960047、82160086);贵州省科学技术基金[编号:黔科合平台人才GCC(2022)040-1]。

摘　　要：目的探讨棕榈油酸(POA)对心肌缺氧/复氧损伤后心肌细胞焦亡的影响。方法实验设置对照组(Control)、缺氧/复氧组(HR)、棕榈油酸处理组(HR+POA)、溶剂无水乙醇(ET)对照组(HR+ET);采用CCK-8法检测心肌细胞活力,Hoechst33342/PI双染和乳酸脱氢酶(LDH)试验检测心肌细胞焦亡水平。ELISA法检测细胞培养上清液中炎症因子白细胞介素(IL)-1β、IL-18释放量,qRT-PCR法和Western blot分别检测细胞焦亡相关分子NLRP3、ASC、Caspase-1、GSDMD、IL-1β及IL-18的mRNA和蛋白相对表达水平。结果CCK-8结果显示,与HR组相比,加入不同浓度POA干预,POA浓度在25~100μmol/L时能增加缺氧/复氧心肌细胞活力;构建心肌细胞缺氧/复氧模型,与对照组相比,HR组中NLRP3、ASC、Cleaved-caspase 1、GSDMD-N、IL-Iβ及IL-18基因的蛋白和mRNA表达增加(P<0.05),Hoechst33342/PI心肌细胞染色阳性百分比增加(P<0.05),且LDH、IL-Iβ及IL-18释放增加(P<0.05);100μmol/L POA干预后,与HR+ET组相比,HR+POA组中NLRP3、ASC、Cleaved-caspase 1、GSDMD-N、IL-Iβ及IL-18的蛋白和mRNA表达降低(P<0.05),Hoechst33342/PI心肌细胞染色阳性百分比降低,且LDH、IL-Iβ及IL-18水平降低(P<0.05)。结论棕榈油酸可能通过抑制缺氧/复氧后心肌细胞焦亡和炎症反应,改善心肌细胞缺氧/复氧损伤。Objective To investigate the effect of palmitoleic acid(POA)on pyroptosis of cardiomyocytes after hypoxia/reoxygenation-induced injury in the human myocardium.Methods The experiment comprised a control group(Control,normal culture),a hypoxia/reoxygenation group(HR),a palmitoleic acid-treated group(HR+POA),and an anhydrous ethanol control group(HR+ET).Cardiomyocytes viability was assessed using CCK-8 assay,and the level of pyroptosis of cardiomyocytes was measured through the double staining with Hoechst33342/PI and LDH assay.ELISA was employed to detect the release of inflammatory factors IL-1βand IL-18 in the cell culture supernatant.qRT-PCR and Western blot were utilized to determine the relative expression levels of mRNA and protein of pyroptosis-related genes,namely NLRP3,ASC,Caspase-1,GSDMD,IL-1βand IL-18,respectively.Results CCK-8 assay showed that the survival of hypoxic/reoxygenated cardiomyocytes increased with the addition of POA at concentrations ranging from 25 to 100μmol/L,as compared to the HR group;a hypoxia/reoxygenation model of cardiomyocyte was established.The expression of protein and mRNA increased in NLRP3,ASC,Cleaved caspase-1,GSDMD-N,IL-Iβand IL-18 vs the control group(P<0.05),the positive percentage of Hoechst33342/PI staining in cardiomyocytes increased significantly(P<0.05),the release of LDH,IL-Iβ,and IL-18 increased(P<0.05).After intervention with 100μmol/L POA,the protein and mRNA expression levels of NLRP3,ASC,Cleaved caspase-1,GSDMD-N,IL-Iβ,and IL-18 were significantly reduced in the HR+POA group vs HR+ET group(P<0.05).The positive percentage of Hoechst33342/PI staining in cardiomyocytes decreased significantly,and the levels of LDH,IL-Iβand IL-18 significantly decreased(P<0.05).Conclusion Palmitoleic acid may alleviate hypoxia/reoxygenation-induced injury of cardiomyocytes by inhibiting pyroptosis and inflammatory response after hypoxia/reoxygenation in human myocardium.

关 键 词：棕榈油酸 心肌细胞 缺氧/复氧损伤 细胞焦亡 炎症 

分 类 号：R541.4[医药卫生—心血管疾病]