乔松素对缺氧/复氧诱导大鼠心肌细胞焦亡的预防作用及其机制  

Pinocembrin inhibits cardiomyocyte pyroptosis induced by hypoxia/reoxygenation in rats

作　　者：张威 卢小伟[1] 许浩[1] ZHANG Wei;LU Xiaowei;XU Hao(Department of Cardiovascular Medicine,Sinopharm Dongfeng General Hospital Affiliated to Hubei Medical College,Shiyan 442008,China)

机构地区：[1]湖北医药学院附属国药东风总医院心血管内科,湖北十堰442008

出　　处：《山东医药》2023年第3期12-16,共5页Shandong Medical Journal

摘　　要：目的观察乔松素(PIN)对缺氧/复氧(H/R)诱导的大鼠心肌细胞焦亡的预防作用,并探讨其预防作用机制。方法将大鼠心肌细胞H9c2分为PIN-H+740-Y-P组、PIN-H组、PIN-L组、模型组、对照组。PIN-H+740-Y-P组用加入25μmol/L PIN+50μg/mL 740-Y-P的培养基、PIN-H组用加入25μmol/L PIN的培养基、PIN-L组用加入12.5μmol/L PIN的培养基、模型组用空白培养基培养1 h之后进行H/R处理。对照组仅用空白培养基培养。取上述各组细胞继续培养24、48、72 h,采用CCK-8法测定细胞活力;取上述各组细胞,采用碘化吡啶(PI)染色法测算各组心肌细胞焦亡比例,ELISA法测定细胞中LDH,Western blotting法检测细胞中磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、核因子-κB(NF-κB)蛋白和细胞焦亡相关蛋白(Caspase-1、ASC和NLRP3蛋白),荧光定量PCR法测定细胞中PI3K、AKT、NF-κB mRNA。结果与对照组相比,模型组细胞活力(48 h、72 h)下降(P均<0.05);焦亡心肌细胞比例,LDH水平,Caspase-1、ASC和NLRP3蛋白相对表达量,PI3K、Akt、NF-κB p65蛋白的磷酸化水平及PI3K、Akt、NF-κB mRNA相对表达量升高(P均<0.05)。与模型组相比,PIN-H组细胞活力(48 h、72 h)上升(P均<0.05);心肌细胞焦亡比例,LDH水平,Caspase-1、ASC和NLRP3蛋白相对表达量,PI3K、Akt、NF-κB p65蛋白的磷酸化水平及PI3K、Akt、NF-κB mRNA相对表达量下降(P均<0.05)。与PIN-H组比较,PIN-H+740-Y-P组细胞活力降低,心肌细胞焦亡比例,LDH水平,Caspase-1、ASC、NLRP3蛋白相对表达量,PI3K、AKT、NF-κB p65蛋白磷酸化水平及PI3K、AKT和NF-κB mRNA相对表达量升高(P均<0.05)。结论PIN对H/R诱导的心肌细胞焦亡有预防作用,其作用机制可能是通过下调PI3K/AKT/NF-κB信号通路实现。Objective To observe the preventive effect of pinocembrin(PIN)on myocardial cell death induced by hypoxia/reoxygenation(H/R)in rats,and to explore its preventive mechanism.Methods The rat cardiomyocytes H9c2 were divided into PIN-H+740-Y-P group,PIN-H group,PIN-L group,model group,and control group.H9c2 cells in the PIN-H+740-Y-P group were incubated with medium including 25μmol/L PIN and 50μg/mL 740-Y-P,PIN-H group with medium including 25μmol/L PIN,PIN-L group with medium including 12.5μmol/L PIN and model group with blank medium for 1 h and then followed by H/R treatment.H9c2 cells in the control group were only cultured with blank medium.The cells of the above groups were cultured for 24,48 and 72 h,and the cell viability was determined by CCK-8.We took the cells of the above groups,measured the proportion of myocardial cells in each group by pyridine iodide(PI)staining,and measured the LDH in the cells by ELISA,and detected the phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and nuclear factor-κB(NF-κB)protein,and pyrolytic related proteins[Caspase-1,apoptosis-associated speck-like protein containing CARD(ASC)and NOD-like receptor protein 3(NLRP3)proteins]in the cells by Western blotting.PI3K,AKT and NF-κB mRNA in cells were determined by fluorescence quantitative PCR.Results Compared with the control group,the cell viability(at 48 and 72 h)decreased(P<0.05),but the cardiomyocyte pyroptosis ratio,LDH level,PI3K,Akt and NF-κB mRNA expression levels,PI3K,Akt,NF-κB p65 protein phosphorylation levels,Caspase-1,ASC and NLRP3 protein expression levels increased in the H/R group(all P<0.05).Compared with the model group,the cell viability increased(at 48 and 72 h)(P<0.05),but the proportion of myocardial cell death,the level of LDH,the relative expression levels of Caspase-1,ASC and NLRP3 proteins,the phosphorylation levels of PI3K,Akt and NF-κB p65 proteins and the relative expression levels of PI3K,Akt and NF-κB mRNA decreased in the PIN-H group(all P<0.05).Compared with the PIN-H group,the ce

关 键 词：乔松素 细胞焦亡 心肌细胞焦亡 磷脂酰肌醇3-激酶/蛋白激酶B/核因子-κB信号通路 缺氧/复氧 

分 类 号：R364.1[医药卫生—病理学]