κ-阿片受体激动剂对缺血/再灌注大鼠心肌及线粒体的保护作用
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摘要
目的探讨κ-阿片受体(κ-OR)激动剂U50488H对大鼠缺血/再灌注(I/R)心肌线粒体的保护作用。方法 20只8周龄雄性SD大鼠被随机分成假手术组(sham)、缺血/再灌注组(I/R)、κ-OR激动剂组(I/R+U50488H)、κ-OR阻断剂(norBNI)组(I/R+N+U50488H),每组5只。比色法检测大鼠血清CK和LDH活力,TUNEL染色观察大鼠心肌细胞凋亡,透射电镜观察大鼠心肌线粒体的形态,Western blot法检测胞质和线粒体内细胞色素C的含量。结果与sham组相比较,I/R组血清CK和LDH显著增加、心肌细胞凋亡率增加、线粒体空泡化增加、线粒体平均面积减小(均P <0.01),线粒体内部细胞色素C释放到胞质的量更大(P <0.01);与I/R相比较,I/R+U50488H组血清CK和LDH降低(P <0.01),心肌细胞凋亡率降低(P <0.05),线粒体空泡化下降(P <0.01),线粒体平均面积增加(P <0.05),线粒体释放色素C减少(P <0.01);与I/R+U50488H组相比较,I/R+N+U50488H组血清CK和LDH增加、心肌细胞凋亡率增加(均P <0.05),线粒体空泡化增加(P <0.01),线粒体平均面积减小(P <0.05),线粒体释放细胞色素C增加(P <0.05)。结论激活κ-OR可以通过改善心肌线粒体的形态与功能,减轻大鼠心肌缺血/再灌注损伤。
Objective To investigate the protective effects of kappa-opioid receptor(κ-OR) agonist on myocardium and mitochondria in ischemia/reperfusion(I/R) rats. Methods Twenty male Sprague-Dawley rats of 8-week-old were randomly divided into four groups(n = 5 in each group) : sham operation group(sham),ischemia/reperfusion group(I/R),κ-OR agonist group(I/R +U50488H),and κ-OR blocker(nor-BNI) group(I/R + N + U50488H). The serum CK and LDH activities were detected by colorimetric assay. The apoptosis of rat cardiomyocytes was detected by TUNEL staining. The morphology of myocardial mitochondria was observed under transmission electron microscope. The content of cytochrome C in cytoplasm and mitochondria was detected by Western blot. Results Compared with sham group,serum CK and LDH,myocardial apoptosis rate,mitochondrial vacuolization and cytochrome C in the cytoplasm were significantly increased in I/R group,while the mitochondrial average area was decreased(all P <0.01). Compared with I/R group,serum CK and LDH decreased in I/R + U50488H group(P < 0.01),myocardial apoptosis rate was decreased(P < 0.05),mitochondrial vacuolization was decreased(P < 0.01),cytochrome C in the cytoplasm was decreased(P <0.01),while the mitochondrial average area increased(P < 0.05). Compared with I/R + U50488H group,serum CK and LDH levels,cardiomyocyte apoptosis rate,and cytochrome C in the cytoplasm increased in I/R + N + U50488H group(all P < 0.05),and mitochondrial vacuolization was also increased(P < 0.01),while the average area of mitochondria decreased(P < 0.05). Conclusion Activation of κ-OR can alleviate myocardial I/R injury in rats by improving mitochondrial function and regulating mitochondrial morphology.
Objective To investigate the protective effects of kappa-opioid receptor(κ-OR) agonist on myocardium and mitochondria in ischemia/reperfusion(I/R) rats. Methods Twenty male Sprague-Dawley rats of 8-week-old were randomly divided into four groups(n = 5 in each group) : sham operation group(sham),ischemia/reperfusion group(I/R),κ-OR agonist group(I/R +U50488H),and κ-OR blocker(nor-BNI) group(I/R + N + U50488H). The serum CK and LDH activities were detected by colorimetric assay. The apoptosis of rat cardiomyocytes was detected by TUNEL staining. The morphology of myocardial mitochondria was observed under transmission electron microscope. The content of cytochrome C in cytoplasm and mitochondria was detected by Western blot. Results Compared with sham group,serum CK and LDH,myocardial apoptosis rate,mitochondrial vacuolization and cytochrome C in the cytoplasm were significantly increased in I/R group,while the mitochondrial average area was decreased(all P <0.01). Compared with I/R group,serum CK and LDH decreased in I/R + U50488H group(P < 0.01),myocardial apoptosis rate was decreased(P < 0.05),mitochondrial vacuolization was decreased(P < 0.01),cytochrome C in the cytoplasm was decreased(P <0.01),while the mitochondrial average area increased(P < 0.05). Compared with I/R + U50488H group,serum CK and LDH levels,cardiomyocyte apoptosis rate,and cytochrome C in the cytoplasm increased in I/R + N + U50488H group(all P < 0.05),and mitochondrial vacuolization was also increased(P < 0.01),while the average area of mitochondria decreased(P < 0.05). Conclusion Activation of κ-OR can alleviate myocardial I/R injury in rats by improving mitochondrial function and regulating mitochondrial morphology.
引文
[1]Hausenloy DJ,Yellon DM.Myocardial ischemia-reperfusion injury:a neglected therapeutic target[J].J Clin Invest,2013,123(1):92-100.
[2]Wang Y,Yuan Y,Wang X,et al.Tilianin post-conditioning attenuates myocardial ischemia/reperfusion injury via mitochondrial protection and inhibition of apoptosis[J].Med Sci Monit,2017,23:4490-4499.
[3]Ding M,Ning J,Feng N,et al.Dynamin-related protein 1-mediated mitochondrial fission contributes to post-traumatic cardiac dysfunction in rats and the protective effect of melatonin[J].JPineal Res,2018,64(1).
[4]王英,刘敏,于波涛,等.线粒体乙醛脱氢酶影响心肌缺血再灌注损伤大鼠心肌线粒体功能和室性心动过速的机制[J].山西医科大学学报,2019,50(1):1-6.
[5]Tong G,Sun Z,Wei X,et al.U50488H postconditioning reduces apoptosis after myocardial ischemia and reperfusion[J].Life Sci,2011,88(1-2):31-38.
[6]Shi W,Vinten-Johansen J.Endogenous cardioprotection by ischaemic postconditioning and remote conditioning[J].Cardiovasc Res,2012,94(2):206-216.
[7]Tanaka K,Kersten JR,Riess ML.Opioid-induced cardioprotection[J].Curr Pharm Des,2014,20(36):5696-5705.
[8]Srisakuldee W,Makazan Z,Nickel BE,et al.The FGF-2-triggered protection of cardiac subsarcolemmal mitochondria from calcium overload is mitochondrial connexin 43-dependent[J].Cardiovasc Res,2014,103(1):72-80.
[9]Anderson RI,Becker HC.Role of the dynorphin/kappa opioid receptor system in the motivational effects of ethanol[J].Alcohol Clin Exp Res,2017,41(8):1402-1418.
[10]Zhou Y,Wang Y,Wang X,et al.The protective effects of kappa-opioid receptor stimulation in hypoxic pulmonary hypertension involve inhibition of autophagy through the AMPK-MTOR pathway[J].Cell Physiol Biochem,2017,44(5):1965-1979.
[11]Zhang QY,Wang W,Shi QX,et al.Antiarrhythmic effect mediated by kappa-opioid receptor is associated with Cx43 stabilization[J].Crit Care Med,2010,38(12):2365-2376.
[12]Ding M,Feng N,Tang D,et al.Melatonin prevents Drp1-mediated mitochondrial fission in diabetic hearts through SIRT1-PGC1αpathway[J].J Pineal Res,2018,65(2):e12491.
[13]Paradis S,Charles AL,Meyer A,et al.Chronology of mitochondrial and cellular events during skeletal muscle ischemia-reperfusion[J].Am J Physiol Cell Physiol,2016,310(11):C968-C982.
[14]Leucker TM,Bienengraeber M,Muravyeva M,et al.Endothelial-cardiomyocyte crosstalk enhances pharmacological cardioprotection[J].J Mol Cell Cardiol,2011,51(5):803-811.
[2]Wang Y,Yuan Y,Wang X,et al.Tilianin post-conditioning attenuates myocardial ischemia/reperfusion injury via mitochondrial protection and inhibition of apoptosis[J].Med Sci Monit,2017,23:4490-4499.
[3]Ding M,Ning J,Feng N,et al.Dynamin-related protein 1-mediated mitochondrial fission contributes to post-traumatic cardiac dysfunction in rats and the protective effect of melatonin[J].JPineal Res,2018,64(1).
[4]王英,刘敏,于波涛,等.线粒体乙醛脱氢酶影响心肌缺血再灌注损伤大鼠心肌线粒体功能和室性心动过速的机制[J].山西医科大学学报,2019,50(1):1-6.
[5]Tong G,Sun Z,Wei X,et al.U50488H postconditioning reduces apoptosis after myocardial ischemia and reperfusion[J].Life Sci,2011,88(1-2):31-38.
[6]Shi W,Vinten-Johansen J.Endogenous cardioprotection by ischaemic postconditioning and remote conditioning[J].Cardiovasc Res,2012,94(2):206-216.
[7]Tanaka K,Kersten JR,Riess ML.Opioid-induced cardioprotection[J].Curr Pharm Des,2014,20(36):5696-5705.
[8]Srisakuldee W,Makazan Z,Nickel BE,et al.The FGF-2-triggered protection of cardiac subsarcolemmal mitochondria from calcium overload is mitochondrial connexin 43-dependent[J].Cardiovasc Res,2014,103(1):72-80.
[9]Anderson RI,Becker HC.Role of the dynorphin/kappa opioid receptor system in the motivational effects of ethanol[J].Alcohol Clin Exp Res,2017,41(8):1402-1418.
[10]Zhou Y,Wang Y,Wang X,et al.The protective effects of kappa-opioid receptor stimulation in hypoxic pulmonary hypertension involve inhibition of autophagy through the AMPK-MTOR pathway[J].Cell Physiol Biochem,2017,44(5):1965-1979.
[11]Zhang QY,Wang W,Shi QX,et al.Antiarrhythmic effect mediated by kappa-opioid receptor is associated with Cx43 stabilization[J].Crit Care Med,2010,38(12):2365-2376.
[12]Ding M,Feng N,Tang D,et al.Melatonin prevents Drp1-mediated mitochondrial fission in diabetic hearts through SIRT1-PGC1αpathway[J].J Pineal Res,2018,65(2):e12491.
[13]Paradis S,Charles AL,Meyer A,et al.Chronology of mitochondrial and cellular events during skeletal muscle ischemia-reperfusion[J].Am J Physiol Cell Physiol,2016,310(11):C968-C982.
[14]Leucker TM,Bienengraeber M,Muravyeva M,et al.Endothelial-cardiomyocyte crosstalk enhances pharmacological cardioprotection[J].J Mol Cell Cardiol,2011,51(5):803-811.