葛根素抑制再灌注大鼠心肌线粒体通透性转换孔开放对心肌细胞凋亡和自噬的影响
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摘要
目的观察葛根素抑制心肌缺血/再灌注大鼠线粒体通透性转换孔对心肌细胞凋亡和自噬的影响。方法将40只SPF级SD大鼠随机分成假手术组,MIRI模型组,环孢霉素阳性对照组,葛根素组,每组10只;阻断大鼠升主动脉造成心肌缺血30 min,再灌注120 min造成大鼠心肌缺血再灌注损伤(Myocardial Ischemia reperfusion injury,MIRI)模型,采用环孢霉素和葛根素在结扎后5 min注射给药,测定各组心肌梗死面积,观察药物干预心肌线粒体通透性转换孔(mitochondrial permeability transition pore,m PTP)后对细胞色素C的释放、心肌细胞凋亡及自噬标志物LC3 II、Beclin1的影响。结果与MIRI模型组比较,葛根素组可以减少心肌梗死面积,抑制线粒体m PTP开放,减少细胞色素C水平和细胞凋亡指数,还可以降低自噬标志物LC3 II、Beclin1的表达(P<0.05),药理效果基本与环孢霉素阳性对照药类似。结论葛根素可以减少大鼠心肌缺血/再灌注梗死面积,其机制可能与抑制m PTP的开放,减少再灌注期心肌细胞大量凋亡和自噬有关。
Objective To explore the interfering effect of puerarin on opening of mitochondrial permeability transition pore( m PTP),myocardial cell apoptosis and autophagy in rats with ischemia-reperfusion injury. Methods Forty SPF level healthy male SD rats were randomly divided into four groups( ten in each),the sham operation group,MIRI model group,cephalosporins positive control group,puerarin group. MIRI rat model was carried out by ligation of left anterior descending coronary artery for 30 min followed by reperfusion for 120 min. Cephalosporins and puerarin injection were administered 5 min after the ligation. myocardial infarct size was also estimated. The opening of mitochondrial permeability transition pore and the expression of cytochrome C( Cyt-C),cell apoptosis,LC3 II and Beclin1 were examined after treated. Results Compared with the MIRI model group,puerarin could reduce the myocardial infarct size,inhibit the opening of mitochondrial permeability transition pore,reduce Cyt-C and myocardial apoptosis,reduce the express of LC3 II,Beclin1( P < 0. 05). Effect of puerarin was similar to that of cephalosporins positive drug. Conclusion Puerarin could reduce the myocardial infarct size in the treatment of myocardial ischemiareperfusion injury in rats and its mechanism might be related to inhibitory effect of the MPTP opening,reduce the myocardial cell apoptosis and autophagy on reperfusion period.
Objective To explore the interfering effect of puerarin on opening of mitochondrial permeability transition pore( m PTP),myocardial cell apoptosis and autophagy in rats with ischemia-reperfusion injury. Methods Forty SPF level healthy male SD rats were randomly divided into four groups( ten in each),the sham operation group,MIRI model group,cephalosporins positive control group,puerarin group. MIRI rat model was carried out by ligation of left anterior descending coronary artery for 30 min followed by reperfusion for 120 min. Cephalosporins and puerarin injection were administered 5 min after the ligation. myocardial infarct size was also estimated. The opening of mitochondrial permeability transition pore and the expression of cytochrome C( Cyt-C),cell apoptosis,LC3 II and Beclin1 were examined after treated. Results Compared with the MIRI model group,puerarin could reduce the myocardial infarct size,inhibit the opening of mitochondrial permeability transition pore,reduce Cyt-C and myocardial apoptosis,reduce the express of LC3 II,Beclin1( P < 0. 05). Effect of puerarin was similar to that of cephalosporins positive drug. Conclusion Puerarin could reduce the myocardial infarct size in the treatment of myocardial ischemiareperfusion injury in rats and its mechanism might be related to inhibitory effect of the MPTP opening,reduce the myocardial cell apoptosis and autophagy on reperfusion period.
引文
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[3]Galagudza M,Korolev D,Postnov V,et al.Passive targeting of ischemic-reperfused myocardium with adenosine-loaded silica nanoparticles[J].Int J Nanomedicine,2012,7:1671.
[4]吴爱明,娄利霞,张冬梅,等.益气活血药对心肌梗死大鼠心室缝隙连接蛋白43表达及室颤发生的影响[J].中西医结合心脑血管病杂志,2015,13(16):1837.
[5]陈亚妮,高俊杰,朱文叶,等.速效救心丸干预心肌缺血/再灌注大鼠线粒体通透性转换孔及心肌细胞凋亡[J].中成药,2015,37(3):469.
[6]Wei SY,Chen Y,Xu XY.Progress on the pharmacological research of puerarin:a review[J].Chin J Nat Med,2014,12(6):407.
[7]汪群红,章灵芝,徐文伟,等.葛根素的药理作用与不良反应分析[J].中华中医药学刊,2015,33(5):1185.
[8]张玮玮,郭永清.大鼠急性心肌缺血再灌注模型的改进[J].山西医药杂志,2009,38(10):902.
[9]王士生,黄进宇,杨竞妍,等.辛伐他汀抑制大鼠心肌缺血再灌注时线粒体通透性转换孔开放的研究[J].中华全科医学,2014,12(1):18.
[10]Yang X,Zhao Y,Wu H,et al.The coexistence of comorbidities at admission is an independent predictor of 30-day mortality of patients hospitalized with acute myocardial infarction:analysis of 5523 cases in China[J].Int J Cardiol,2012,155(3):451.
[11]王鹏,马军军,杜亚明.黄芩苷对再灌注心肌细胞保护作用及与心肌细胞自噬的相关性[J].中国循环杂志,2016,31(7):701.
[12]Stowe DF,Camara AKS.Mitochondrial reactive oxygen species production in excitable cells:modulators of mitochondrial and cell function[J].Antioxid Redox Signal,2009,11(6):1373.
[13]Petrosillo G,Di Venosa N,Moro N,et al.In vivo hyperoxic preconditioning protects against rat-heart ischemia/reperfusion injury by inhibiting mitochondrial permeability transition pore opening and cytochrome c release[J].Free Radic Biol Med,2011,50(3):477.
[14]Ma S,Wang YB,Chen YD,et al.The role of the autophagy in myocardial ischemia/reperfusion injury[J].Biochim Biophys Acta,2015,2(1852):271.
[15]Gao Q,Pan HY,Qiu S,et al.Atractyloside and 5-hydroxydecanoate block the protective effect of puerarin in isolated rat heart[J].Life Sci,2006,79(3):217.
[16]姚慧,高琴,夏强.线粒体钾通道参与葛根素抗心肌细胞缺氧/复氧损伤的作用[J].中国应用生理学杂志,2010,26(4):459.
[17]Jian J,Xuan FF,Qin FZ,et al.Bauhinia championii flavone inhibits apoptosis and autophagyvia the PI3K/Akt pathway in myocardialischemia/reperfusion injury in rats[J].Drug Des Devel Ther,2015,9:5933.