羟基红花黄色素A抗心肌细胞缺氧复氧损伤的线粒体相关机制

Mitochondrial mechanism of cardioprotective effect of hydroxysafflor yellow A against anoxia /reoxygenation injury in rats

目的:探讨羟基红花黄色素A(hydroxysafflor yellow A,HSYA)能否通过阻止线粒体通透性转换孔的开放从而保护心肌缺氧复氧损伤以及一氧化氮(NO)在线粒体机制中的作用。方法:采用酶解法分离大鼠心肌细胞模型,台盼蓝拒染法测定心肌细胞存活率;荧光染料四甲基罗丹明乙酯测定线粒体膜电位,分离线粒体测定其通透性转换孔开放程度。结果:HSYA(0.1 mmol/L)预处理5 min可明显提高缺氧复氧心肌细胞的存活率,对抗缺氧复氧引起的线粒体膜电位的去极化。在分离心肌线粒体模型上,HSYA(0.1 mmol/L,5 min)显著减弱CaCl2诱导的线粒体肿胀。一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸甲酯减弱了HSYA对缺氧复氧心肌细胞、线粒体肿胀和对线粒体膜去极化的保护作用。结论:HSYA预处理具有抗缺氧复氧损伤的作用,这种保护作用可能与其抑制线粒体通透性转换孔的开放有关。NO机制参与了HSYA的抗缺氧复氧损伤的保护作用。

Objective： To determine whether the cardioprotection of hydroxysafflor yellow A（HSYA） against anoxia / reoxygenation injury is mediated by mitochondrial transmembrane pores.Method： Cardiomyocytes were isolated from male Sprague-Dawley rats.Cell viability was assessed by trypan blue exclusion and mitochondrial membrane potential was measured by loading with TMRE.The opening of mitochondrial permeability transition pore was determined spectrophotometrically.Result： Pretreatment with HSYA at 0.1 mmol / L for 5 min increased the cell viability against anoxia / reoxygenation injury and attenuated mitochondrial depolarization induced by anoxia / reoxygenation.In mitochondrial isolated from hearts pretreated with 0.1 mmol / L HSYA for 5 min,a significant inhibition of Ca2 ＋ induced swelling was observed,while NG-nitro-L-argininemethyl ester（L-NAME）,an inhibitor of NOS,abrogated the protective effects of HSYA.Conclusion： These findings indicate that HSYA protected cardiomyocytes against anoxia /reoxygenation injury via inhibited mitochondrial permeability transition pore opening,while nitric oxide may take a part in it.