益心舒对急性心肌梗死小鼠的保护作用及机制研究

Protective effect of Yixinshu on mice with acute myocardial infarction and its mechanisms

目的:探究益心舒(YXS)对急性心肌梗死(MI)小鼠的心脏保护作用及其相关机制。方法:联合TCMSP与Metascape数据库进行生物信息学分析,探寻YXS可能的作用靶点。将60只雄性C57BL/6J小鼠随机分为3组:假手术组(sham组)、模型组(MI组)和YXS预处理组(YXS+MI组),每组各20只。YXS+MI组给予YXS(60 mg·kg^(−1)·d^(−1))灌胃,连续14 d,sham组和MI组给予等体积生理盐水灌胃。通过结扎冠状动脉左前降支建立MI模型,术后1 d用超声多普勒检测小鼠心功能;通过TTC染色检测MI面积;通过HE染色和CD45免疫组化染色分析梗死部位炎症细胞的浸润情况;采用透射电镜观察心肌超微结构和线粒体损伤;TUNEL染色检测凋亡细胞比例;Western blot检测凋亡相关蛋白Bax和Bcl2的蛋白水平;流式细胞术检测髓源性炎症细胞的浸润情况。结果:与sham组比较,MI组小鼠左心收缩功能受损,射血分数(EF)和短轴缩短率(FS)也显著下降(P<0.01),心肌组织炎症细胞浸润增多,心肌超微结构受到破坏,凋亡细胞比例显著增加(P<0.01),Bax蛋白含量显著增高(P<0.01),Bcl2蛋白显著降低(P<0.01),流式结果显示髓源性炎症细胞浸润显著增加(P<0.01)。与MI组比较,YXS+MI组EF和FS显著上升(P<0.05),心肌组织炎症细胞浸润减少,心肌显微结构损伤减轻,凋亡细胞比例显著降低,Bax蛋白表达减少(P<0.05),而Bcl2蛋白表达增多(P<0.01),髓源性炎症细胞浸润显著减少(P<0.05)。结论:YXS预处理能够减轻小鼠急性MI后的心肌损伤,减少心肌细胞凋亡并降低髓源性炎症细胞的浸润。

AIM:To explore the effect of Yixinshu(YXS)tablets on acute myocardial infarction(MI)in mice and its related mechanism.METHODS:The potential targets of YXS were explored by combining TCMSP and Metascape for bioinformatics analysis.Sixty male C57BL/6J mice were randomly divided into 3 groups:sham operation group(sham group),model group(MI group),and Yixinshu tablet pretreatment group(YXS+MI group),with 20 mice in each group.The mice in YXS+MI group were given YXS(60 mg·kg^(−1)·d^(−1))intragastric administration for 14 d,while those in sham group and MI group were given the same volume of normal saline.The MI model was established by ligation of the left anterior descending coronary artery.The cardiac function was detected by ultrasonic Doppler 1 d after operation.Myocardial infarction size was detected by TTC staining.The infiltration of inflammatory cells at the infarct myocardium was analyzed by HE staining and CD45 immunohistochemistry.Myocardial ultrastructures and mitochondrial damage were observed by transmission electron microscopy.TUNEL staining was used to detect the apoptotic cells.Western blot were used to detect the levels of apoptosis-related proteins Bax and Bcl2,and flow cytometry was used to detect the infiltration of myeloid inflammatory cells.RESULTS:Compared with sham group,the left ventricular systolic function of the mice in MI group was impaired,with significant decreases in ejection fraction(EF)and shortened fraction(FS)(P<0.05).In addition,the inflammatory cell infiltration in myocardial tissue increased,myocardial ultrastructure was damaged,the proportion of apoptotic cells significantly increased(P<0.01),Bax protein level significantly increased(P<0.01),and Bcl2 protein level significantly decreased(P<0.01).Flow cytometry results showed a significant increase in the infiltration of myeloid inflammatory cells(P<0.01).Compared with MI group,the mice in YXS+MI group showed significant increases in EF and FS(P<0.05).Meanwhile,the infiltration of inflammatory cells in myocardial tissues was reduced,and the damage to myocardial ultrastructures was alleviated,the proportion of apoptotic cells significantly decreased(P<0.01),with a decrease in Bax protein expression(P<0.05),an increase in Bcl2 protein expression(P<0.01)and a reduction of myeloid inflammatory cell infiltration(P<0.05).CONCLUSION:Pretreatment with YXS alleviates myocardial injury after acute MI in mice,reduces cardiomyocyte apoptosis,and decreases the infiltration of myeloid inflammatory cells.