芦丁对大鼠心脏缺血再灌注损伤的影响

Influence of Rutin on Myocardial Ischemia-Reperfusion Injury

关键词:  芦丁  心脏缺血再灌注损伤  炎症  氧化应激

Key words:  rutin  myocardial ischemia reperfusion injury  inflammation  oxidative stress

摘要:

目的 探讨芦丁对心脏缺血再灌注损伤大鼠的影响及机制。方法 40只SD大鼠随机分为假手术组（Sham）、心脏缺血-再灌注损伤组（IRI）和芦丁组群（Rutin）。IRI组和Rutin组无损伤血管夹夹闭冠状动脉左前降支0.5 h构建IRI模型，Sham组只做假手术处理。Rutin组群在缺血前0.5 h腹腔分别注射Rutin（20，40 mg·kg^-1），Sham组和IRI组腹腔注射等量生理盐水。再灌注4 h后，收集各组大鼠心脏和血清。苏木精-伊红染色法检测各组大鼠心脏病理形态；ELISA检测血清中肌酸激酶同工酶（CK-MB），乳酸脱氢酶（LDH），巨噬细胞炎症因子1（MCP-1），白介素6（IL-6）和肿瘤坏死因子-α（TNF-α）的表达；Western blotting检测心脏磷脂酰肌醇3-激酶（PI3K），蛋白激酶B（AKT）、磷酸化蛋白激酶B（p-AKT）和雷帕霉素靶蛋白（mTOR）的表达；黄嘌呤氧化酶法检测心脏过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GPx）和超氧化物歧化酶（SOD）活性；硫代巴比妥酸法检测心脏丙二醛（MDA）含量；RT-PCR检测心脏MCP-1，IL-6和TNF-α的mRNA表达。结果 与Sham组相比，IRI组心脏病理改变明显增加，血清中CK-MB、LDH、MCP-1、IL-6、和TNF-α表达明显增加，心脏PI3K、p-AKT、mTOR、MDA、MCP-1、IL-6和TNF-α表达明显增加，而CAT、GP_X与SOD活性明显减少。与IRI组相比，Rutin组心脏病理改变明显减少，血清中CK-MB、LDH、MCP-1、IL-6、和TNF-α表达明显减少，心脏PI3K、p-AKT、mTOR、MDA、MCP-1、IL-6和TNF-α表达明显减少，而CAT、GP_X与SOD活性明显增加。此外，3组之间AKT表达无统计学差异。结论 芦丁预处理可呈剂量依赖性减轻心脏缺血再灌注损伤，其作用机制部分是通过激活PI3K/AKT/mTOR信号通路抑制炎症和氧化应激。

Abstract:

OBJECTIVE To explore the effect and mechanism of rutin on myocardial ischemia reperfusion injury in rats. METHODS Forty SD rats were randomly divided into Sham group(Sham), myocardial ischemia reperfusion injury group(IRI) and Rutin groups(Rutin). IRI and Rutin groups were induced IRI by clamping the left anterior descending coronary artery for 0.5 h with noninvasive endoclips. Sham group with sham operation. Rutin groups were pretreated with Rutin(20, 40 mg·kg^-1) at 0.5 h before ischemia by intraperitoneal injection. Rats in IRI group and Sham group were administrated with same dosage saline at the same time. After reperfusion for 4 h, the cardiac tissue and serum were collected. The cardiac pathology was exanimed by HE; The expression of CK-MB, LDH, MCP-1, IL-6, TNF-α in serm were evaluated by ELISA; The expression of PI3K, AKT, p-AKT and m-TOR in heart were evaluated by Western blotting; The expression of MDA and the activity of CAT, GP_X and SOD in heart were examined by TBA and xanthine oxidase method. The expression of IL-6, MCP-1 and TNF-α in serum and cardial tissue were determined by ELISA and RT-PCR. RESULTS Compared with the Sham group, the cardiac morphological change and the expression of CK-MB, LDH, MCP-1, IL-6, and TNF-α in serum, PI3K, p-AKT, mTOR, MDA, MCP-1, IL-6 and TNF-α in heart in IRI group were markedly increased with decreasing the activity of CAT, GP_X and SOD in IRI group. Compared with the IRI group, the cardiac morphological change and the expression of CK-MB, LDH, MCP-1, IL-6, and TNF-α in serum, PI3K, p-AKT, mTOR, MDA, MCP-1, IL-6 and TNF-α in heart were decreased with increasing the activity of CAT, GP_X and SOD in Rutin group. There was no difference on the expression of AKT among three groups. CONCLUSION Rutin can protect rats against myocardial ischemia-reperfusion injury. The mechanism of which is partly by suppressing inflammation and Oxidative stress though promoting the activation of PI3K/AKT/mTOR signal pathway.