槲皮素基于PI3K/AKT/GSK-3<i>β</i>/<i>β</i>-Catenin通路改善Dox诱导小鼠心肌损伤的机制

张珊; 高志辉; 王秋红; 李昊泽; 姜宏锦; 张红军

doi:10.13386/j.issn1002-0306.2023120038

槲皮素基于PI3K/AKT/GSK-3β/β-Catenin通路改善Dox诱导小鼠心肌损伤的机制

The Mechanism of Quercetin Improving Dox-induced Myocardial Injury in Mice Based on PI3K/AKT/GSK-3β/β-Catenin Pathway

摘要

摘要: 目的：研究槲皮素（Que）改善多柔比星（Dox）所致小鼠心肌损伤作用机制。方法：将50只小鼠随机分为正常组（Control）、模型组（Dox）、槲皮素低、中、高剂量组（Que-L、Que-M、Que-H），除Control外腹腔注射Dox建立心肌损伤模型。心脏超声及血流动力学评价小鼠心脏功能，ELISA法检测小鼠血清中肌酸激酶同工酶（CK-MB）与乳酸脱氢酶（LDH）含量，HE染色、Tunel染色、WGA染色分别观察小鼠心肌组织病理变化、心肌细胞凋亡、心肌细胞横截面积变化；免疫荧光检测心肌组织中p-GSK-3β表达，Western blot法检测小鼠心肌组织PI3K/AKT/GSK-3β通路与凋亡蛋白表达。结果：与Control组比较，Dox组小鼠左室射血分数（LVEF）与左室短轴缩短率（LVFS）显著下降，左心室舒张末期内径（LVEDd）与左心室收缩末期内径（LVEDs）显著增加，血清中CK-MB与LDH含量极显著增加（P<0.01），心肌细胞肿胀且排列紊乱；心肌组织中PI3K、p-AKT、p-GSK-3β、β-catenin蛋白极显著降低（P<0.01），Bax/Bcl-2、Cleaved Caspase-3表达极显著增加（P<0.01）。与Dox组比较，Que各给药组小鼠心肌损伤均有不同程度改善，LVEF与LVFS极显著升高（P<0.01），（LVEDd）与（LVEDs）极显著降低，血清中CK-MB、LDH含量极显著减少（P<0.01），心脏功能增强；心肌细胞肿胀、凋亡、纤维增生减轻，心肌组织中PI3K、p-AKT、p-GSK-3β、β-catenin蛋白表达极显著升高（P<0.01），PI3K/AKT/GSK-3β/β-catenin被激活，凋亡蛋白Bax/Bcl-2、Cleaved Caspase-3表达极显著降低（P<0.01），其中Que-H组治疗效果最佳。结论：Que具有一定的心肌保护作用，可通过激活PI3K/AKT/GSK-3β/β-catenin通路改善Dox引起的心肌损伤与凋亡。

Abstract: Objective: To study the mechanism of quercetin (Que) in improving myocardial injury induced by doxorubicin (Dox) in mice. Methods: Fifty mice were randomly divided into normal group (Control), model group (Dox), quercetin low-dose, medium-dose and high-dose groups (Que-L, Que-M and Que-H), and myocardial injury model was established by intraperitoneal injection of Dox in addition to Control. The cardiac function of mice was evaluated by use echocardiography and hemodynamics, and creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) levels were detected in serum of mice by ELISA. HE staining, Tunel staining and WGA staining respectively were used to observe the pathological changes of myocardial tissue, myocardial cell apoptosis and myocardial cell cross-sectional area. The expression of p-GSK-3β in myocardial tissue was detected by immunofluorescence, and the expression of PI3K/AKT/GSK-3β pathway and apoptotic protein in mouse myocardial tissue was detected by Western blot. Results: Compared with Control group, left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) in Dox group were significantly decreased, left ventricular end-diastolic diameter (LVEDd) and left ventricular end-systolic diameter (LVEDs) were significantly increased, and serum CK-MB and LDH contents were extremely significant increased (P<0.01). Cardiomyocytes were swollen and disordered organization. The proteins of PI3K, p-Akt, p-GSK-3β and β-catenin in myocardial tissue were significantly decreased (P<0.01), and the expressions of Bax/Bcl-2 and Cleaved Caspase-3 were extremely significant increased (P<0.01). Compared with Dox group, the myocardial injury of mice in each of Que administration groups were improved to varying degrees, LVEF and LVFS were significantly increased (P<0.01), LVEDd and LVEDs were significantly decreased, serum levels of CK-MB and LDH contents were significantly decreased (P<0.01), and cardiac function was enhanced. The swelling, apoptosis and fibroplasia of cardiomyocytes were reduced, and the proteins expression of PI3K, p-Akt, p-GSK-3β and β-catenin in myocardial tissue were significantly increased (P<0.01), and PI3K/AKT/GSK-3β/β-catenin was activated. The expression of apoptosis proteins Bax/Bcl-2 and Cleaved Caspase-3 were significantly decreased (P<0.01), and the Que-H group had the best therapeutic effect. Conclusions: Que has a certain myocardial protective effect, and can improve Dox induced myocardial injury and apoptosis by activating PI3K/AKT/GSK-3β/β-catenin pathway.