Huang Xiaoyan, Wang Yuguang, Wang Yi, Yang Liang, Wang Jia, Gao Yue
Tianjin University of Traditional Chinese Medicine, Tianjin, China.
Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing, China.
Cell Physiol Biochem. 2018;49(4):1646-1658. doi: 10.1159/000493500. Epub 2018 Sep 18.
BACKGROUND/AIMS: Epoxyeicosatrienoic acids (EETs) are cytochrome P450 epoxygenase (CYP) metabolites of arachidonic acid and have multiple cardiovascular effects. Ophiopogonin D (OP-D) is an important effective monomeric component in Shenmai injection (SM-I). Both have been reported to have a variety of biological functions, including anti-inflammatory, anti-oxidant, and anti-apoptotic effects. We previously demonstrated that OP-D-mediated cardioprotection involves activation of CYP2J2/3 and enhancement of circulating EETs levels in vitro and can be developed as a novel drug for the therapy of myocardial ischemia-reperfusion (MI/R) injury. We therefore hypothesized that the protective effects of OP-D and SM-I against MI/R injury are associated with increased expression of CYP2J3 and enhanced circulating 11,12-EET levels in vivo.
A rat model of MI/R injury was generated by ligation of the left anterior descending coronary artery for 40 min, followed by reperfusion for 2 h to determine the protective effects and potential mechanisms of OP-D and SM-I. Electrocardiogram and ultrasonic cardiogram were used to evaluate cardiac function; 2,3,5-triphenyltetrazolium chloride was used to measure myocardial infarct size; hematoxylin and eosin staining and transmission electron microscopy were used to observe the morphology of myocardial tissue; and the expression of related proteins in the mechanistic study was observed by western blot analysis.
We found that OP-D and SM-I exert protective effects on MI/R injury, including regulation of cardiac function, reduction of lactate dehydrogenase and creatine kinase production, attenuation of myocardial infarct size, and improvement of the recovery of damaged myocardial structures. We found that OP-D and SM-I activate CYP2J3 expression and increase levels of circulating 11,12-EET in MI/R-injured rats.
We tested the hypothesis that the cardioprotective effects of OP-D and SM-I on MI/R injury are associated with increased expression of CYP2J3 and enhanced circulating 11,12-EET levels in rats. Taken together, our results show that the effects of OP-D and SM-I were also mediated by the activation of the PI3K/Akt/eNOS signaling pathway, while inhibition of the NF-κB signaling pathway and antioxidant and anti-apoptotic effects were involved in the cardioprotective effects of OP-D and SM-I.
背景/目的：环氧二十碳三烯酸（EETs）是细胞色素P450环氧化酶（CYP）催化花生四烯酸生成的代谢产物，具有多种心血管效应。麦冬皂苷D（OP-D）是参麦注射液（SM-I）中一种重要的有效单体成分。二者均已被报道具有多种生物学功能，包括抗炎、抗氧化和抗凋亡作用。我们之前证实，OP-D介导的心脏保护作用在体外涉及CYP2J2/3的激活和循环EETs水平的升高，并且可开发成为一种治疗心肌缺血再灌注（MI/R）损伤的新型药物。因此，我们推测OP-D和SM-I对MI/R损伤的保护作用与体内CYP2J3表达增加及循环11,12-EET水平升高有关。
通过结扎左冠状动脉前降支40分钟，随后再灌注2小时，建立大鼠MI/R损伤模型，以确定OP-D和SM-I的保护作用及潜在机制。采用心电图和超声心动图评估心脏功能；用2,3,5-氯化三苯基四氮唑测量心肌梗死面积；用苏木精-伊红染色和透射电子显微镜观察心肌组织形态；通过蛋白质印迹分析观察机制研究中相关蛋白的表达。
我们发现OP-D和SM-I对MI/R损伤具有保护作用，包括调节心脏功能、减少乳酸脱氢酶和肌酸激酶生成、减轻心肌梗死面积以及改善受损心肌结构的恢复。我们发现OP-D和SM-I可激活MI/R损伤大鼠的CYP2J3表达并提高循环11,12-EET水平。
我们验证了OP-D和SM-I对MI/R损伤的心脏保护作用与大鼠CYP2J3表达增加及循环11,12-EET水平升高有关这一假设。综上所述，我们的结果表明，OP-D和SM-I的作用还通过PI3K/Akt/eNOS信号通路的激活介导，而抑制NF-κB信号通路以及抗氧化和抗凋亡作用参与了OP-D和SM-I的心脏保护作用。
