漆黄素对大鼠体外心肌缺血-再灌注损伤的影响

Effect of Fisetin on Myocardial Ischemia-reperfusion Injury in Isolated Rats

目的探讨漆黄素抗大鼠体外心肌缺血-再灌注(I/R)损伤作用及作用机制。方法通过Langendorff体外心脏灌流系统建立心肌I/R损伤模型,32只SPF级SD大鼠随机分为4组:正常对照组、模型对照组、漆黄素小剂量组(0.5μmol·L-1)和漆黄素大剂量组(5μmol·L-1),每组8只。使用BL420S生物机能实验系统测定心功能参数;通过氯化三苯基四氮唑(TTC)染色检测心肌梗死面积;酶联免疫吸附测定(ELISA)法检验灌流液中肌酸激酶同工酶(CK-MB)含量;ELISA法检测心肌组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的水平;苏木精-伊红(HE)染色法观察心肌组织的变化;通过TUNEL染色检测心肌细胞凋亡程度。结果与正常对照组比较,模型对照组心功能参数显著降低,心肌组织严重梗死和坏死,心肌细胞中CK-MB的漏出量显著升高,心肌组织中MDA,GSSG,IL-6和TNF-α含量显著升高,SOD和GSH的活性显著降低,模型对照组心肌细胞严重凋亡。与模型对照组比较,漆黄素小、大剂量组可以明显逆转I/R诱导的心肌损伤变化,大剂量漆黄素的心肌保护作用更为明显。结论漆黄素可显著减轻I/R诱导的心肌损伤,并具有一定的剂量依赖性;作用机制可能与其抗氧化、抗炎和抗凋亡作用相关。

Objective To investigate the effect of fisetin on myocardial ischemia reperfusion（I/R） injury in isolated rats and its mechanism. Methods The myocardial I/R injury model was established through Langendorff system. A total of 32 SPF grade SD rats were randomly divided into 4 groups（n = 8） ： normal control group,model control group,low dose fisetin group（0.5 μmol·L-1） and high dose fisetin group（5 μmol·L-1）. The myocardial function was measured by TTC staining.The content of CK-MB was measured by enzyme-linked immunosorbent assay（ELISA）. The levels of SOD,MDA,GSH,GSSG,TNF-α and IL-6 in the myocardium were measured by ELISA. The changes of myocardial tissue were observed by HE staining.The degree of apoptosis was detected by TUNEL staining. Results Compared with the normal control group,the cardiac function parameters of the model control group were decreased significantly,serious infarction and necrosis developed in myocardium,and the leaking amount of CK-MB was significantly increased. The contents of MDA,GSSG,IL-6 and TNF-α in myocardium were significantly increased,and the activities of SOD and GSH significantly decreased. Severe apoptosis appeared in the cardiomyocytes of the model control group. Compared with the model control group,the pretreatment of fisetin could significantly reverse the changes of myocardial injury induced by I/R,and the protective effect of high dose of fisetin was more obvious. Conclusion Fisetin can significantly alleviate I/R-induced myocardial injury and exerts a dose-dependent effect. The protective effect of fisetin may be related to its anti-oxidation,anti-inflammatory and anti-apoptotic effects.