Li W-N, Wu N, Shu W-Q, Guan Y-E, Jia D-L
Department of Cardiology, The First Affiliated Hospital of China Medical University, Liaoning, China.
Eur Rev Med Pharmacol Sci. 2014;18(18):2748-58.
Ischemic postconditioning (IPO) and pharmacological pretreatment may reduce myocardial necrosis and apoptosis during ischemia/reperfusion. This study aimed to determine the protective effect of fasudil pretreatment combined with IPO on myocardial ischemia/reperfusion injury in rats and explore the possible mechanisms.
The SD rats were induced by intraperitoneal injection of fasudil hydrochloride (1 or 10 mg/kg) 60 min before the initiation of ischemia, while the control rats were given the same volume of saline. The hearts were hung on the Langendorff perfusion apparatus and underwent 30 min global ischemia and 120 min reperfusion. The IPO protocol was induced by six cycles of 10 sec ischemia and 10 sec reperfusion at the onset of reperfusion. The hemodynamic changes were measured, myocardial infarct size was determined by triphenyltetrazolium chloride (TTC) staining, cardiomyocyte apoptosis was detected by TUNEL staining, lactate dehydrogenase (LDH) was analyzed from coronary effluents, phosphorylation of Akt and eNOS, as well as expression of Bcl-2 and Bax were measured by western blotting analysis.
The high-dose fasudil (10 mg/kg) pretreatment group and IPO group significantly improved post-ischemia cardiac function, reduced myocardial infarct size, attenuated cardiomyocyte apoptosis, decreased the release of LDH, increased expression of phospho-Akt, phospho-eNOS and Bcl-2, and reduced expression of Bax compared with the control group (p < 0.05). In addition, the high-dose fasudil pretreatment combined with IPO group could further improved post-ischemia cardiac function, reduced myocardial infarct size, attenuated cardiomyocyte apoptosis, decreased the release of LDH, increased expression of phospho-Akt, phospho-eNOS and Bcl-2, and reduced expression of Bax compared with the single treatment groups (p < 0.05).
The combination of high-dose fasudil pretreatment and IPO had a synergistic protective effect on myocardial ischemia/reperfusion injury, which was mediated via upregulating the PI3K/Akt/eNOS pathway, increasing expression of antiapoptotic Bcl-2, and decreasing expression of proapoptotic Bax.
缺血后处理（IPO）和药物预处理可减少缺血/再灌注期间的心肌坏死和凋亡。本研究旨在确定法舒地尔预处理联合IPO对大鼠心肌缺血/再灌注损伤的保护作用，并探讨其可能机制。
在缺血开始前60分钟，通过腹腔注射盐酸法舒地尔（1或10mg/kg）诱导SD大鼠，而对照组大鼠给予相同体积的生理盐水。将心脏悬挂在Langendorff灌注装置上，进行30分钟全心缺血和120分钟再灌注。IPO方案在再灌注开始时通过6个周期的10秒缺血和10秒再灌注诱导。测量血流动力学变化，通过氯化三苯基四氮唑（TTC）染色确定心肌梗死面积，通过TUNEL染色检测心肌细胞凋亡，从冠状动脉流出液中分析乳酸脱氢酶（LDH），通过蛋白质印迹分析测量Akt和eNOS的磷酸化以及Bcl-2和Bax的表达。
与对照组相比，高剂量法舒地尔（10mg/kg）预处理组和IPO组显著改善缺血后心功能，减小心肌梗死面积，减轻心肌细胞凋亡，减少LDH释放，增加磷酸化Akt、磷酸化eNOS和Bcl-2的表达，并降低Bax的表达（p<0.05）。此外，与单一治疗组相比，高剂量法舒地尔预处理联合IPO组可进一步改善缺血后心功能，减小心肌梗死面积，减轻心肌细胞凋亡，减少LDH释放，增加磷酸化Akt、磷酸化eNOS和Bcl-2的表达，并降低Bax的表达（p<0.05）。
高剂量法舒地尔预处理与IPO联合对心肌缺血/再灌注损伤具有协同保护作用，其通过上调PI3K/Akt/eNOS途径、增加抗凋亡蛋白Bcl-2的表达以及降低促凋亡蛋白Bax的表达介导。
