Evidence that hydroxysafflor yellow a protects the heart against ischaemia-reperfusion injury by inhibiting mitochondrial permeability transition pore opening
Evidence that hydroxysafflor yellow a protects the heart against ischaemia-reperfusion injury by inhibiting mitochondrial permeability transition pore opening
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DOI：
10.1111/j.1440-1681.2007.04814.x
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发表时间：
2008-02-01
影响因子：
2.9
通讯作者：
Chen, Peng
中科院分区：
文献类型：
--
作者：
Liu, Yi-Na;Zhou, Zhong-Min;Chen, Peng
1.本研究旨在探讨羟色胺黄A（HSYA）对缺血再灌注心肌损伤的保护作用及其可能的机制.将从雄性Sprague-Dawley大鼠分离的心脏在Langendorff装置上灌注，并进行30分钟的全脑缺血，随后进行120分钟的再灌注。测定冠脉流出液中乳酸脱氢酶（LDH）的含量。在离体灌流心脏的线粒体中，观察到Ca 2+诱导的肿胀。在分离的心室肌细胞，线粒体膜的去极化测定四甲基罗丹明乙酯（TMRE）荧光。Western blot检测内皮型一氧化氮合酶（eNOS）蛋白磷酸化水平.缺血前给予HSYA 5 min可减少心肌梗死面积和LDH释放。20 μ mol/L线粒体通透性转换孔开放剂-- 0.1mmol/L HSYA预处理5 min，可明显抑制心肌线粒体Ca ~（2+）诱导的肿胀，L-NAME可减弱这种抑制作用。在离体心室肌细胞中，HSYA预处理可防止缺血诱导的细胞死亡和线粒体膜去极化，而HSYA或L-NAME可减弱HSYA的作用。HSYA处理组eNOS蛋白磷酸化水平显著升高.本研究结果表明，HSYA通过抑制线粒体通透性转换孔开放来保护心肌免受缺血再灌注损伤。HSYA对线粒体通透性转换孔开放的作用可能是通过增强eNOS激活引起的一氧化氮产生来介导的。
1. The present study was conducted to investigate whether hydroxysafflor yellow A (HSYA) has a protective effect against heart injury after ischaemia-reperfusion and to determine the possible mechanism involved.2. Hearts isolated from male Sprague-Dawley rats were perfused on a Langendorff apparatus and subjected to 30 min global ischaemia, followed by 120 min reperfusion. Infarct size and the level of lactate dehydrogenase (LDH) in the coronary effluent were determined. In mitochondria from isolated perfused hearts, Ca2+-induced swelling was observed. In isolated ventricular myocytes, depolarization of the mitochondrial membrane was determined by tetramethyl-rhodamine ethyl ester (TMRE) fluorescence. Furthermore, levels of phosphorylated endothelial nitric oxide synthase (eNOS) protein were measured by western blot.3. Pretreatment with HSYA for 5 min before ischaemia reduced infarct size and the release of LDH. Administration of 20 mu mol/L atractyloside, an opener of the mitochondrial permeability transition pore, and 10 mu mol/L N-G-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NOS, attenuated the protective effects of HSYA. In mitochondria isolated from hearts pretreated with 0.1 mmol/L HSYA for 5 min, a significant inhibition of Ca2+-induced swelling was observed and this inhibition was attenuated by L-NAME. In isolated ventricular myocytes, pretreatment with HSYA prevented ischaemia-induced cell death and depolarization of the mitochondrial membrane, whereas atractyloside or L-NAME attenuated the effects of HSYA. Levels of phosphorylated eNOS protein were significantly enhanced in the HSYA-treated group.4. The findings of the present study indicate that HSYA protects the myocardium against ischaemia-reperfusion injury by inhibiting mitochondrial permeability transition pore opening. The effect of HSYA on mitochondrial permeability transition pore opening may be mediated through enhanced nitric oxide production by eNOS activation.