益心方抑制NLRP3炎症小体介导的细胞焦亡减轻大鼠心肌缺血再灌注损伤  

Yixin Formula Inhibits NLRP3 Inflammasomes Mediated Pyroptosis to Attenuates Myocardial IschemiaReperfusion Injury in Rats

作　　者：杨燕玲 董莉 郑钰凡 戎靖枫[1] YANG Yanling;DONG Li;ZHENG Yufan;RONG Jingfeng(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203

出　　处：《世界科学技术-中医药现代化》2024年第9期2483-2491,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：上海市卫生健康委员会面上项目(202240225):益心方改善气虚血瘀型心肌缺血再灌注损伤的临床研究,负责人:戎靖枫。

摘　　要：目的探讨益心方对心肌缺血再灌注损伤大鼠心肌保护作用的可能机制。方法将65只Wistar大鼠随机分为假手术组10只、造模组55只;造模组采取结扎冠状动脉左前降支方法构建大鼠MIRI模型,假手术组只穿线不结扎。造模组造模后随机分为模型组、益心方低、中、高剂量组和曲美他嗪组。益心方低、中、高剂量组分别于造模后予以益心方6.84 g·kg^(-1)、13.68 g·kg^(-1)、27.36 g·kg^(-1)灌胃,曲美他嗪组予以曲美他嗪5.4 mg·kg^(-1)灌胃,假手术组与模型组予等体积的生理盐水灌胃,各组灌胃7天。超声心电图检测大鼠心功能情况;生化法检测血清肌酸激酶(CK)、乳酸脱氢酶(LDH)、谷草转氨酶(AST)含量;苏木素-伊红(HE)染色观察大鼠心肌组织病理变化;Masson染色观察心肌纤维化程度;TUNEL染色检测心肌细胞焦亡指数;Western blot检测心肌组织NLRP3、pro-Caspase-1、GSDMD-FL、GSDMD-N蛋白表达。结果与假手术组比较,模型组大鼠左室射血分数(LVEF),左室短轴缩短率(FS)显著降低(P<0.001),心肌损伤标志物CK、AST、LDH显著升高(P<0.001),病理结果显示心肌结构受损,间质水肿伴炎性细胞浸润,心肌纤维排列紊乱,心肌细胞横截面积变大(P<0.001),心肌纤维化明显(P<0.001),心肌细胞焦亡指数显著增加(P<0.001),心肌组织中NLRP3、pro-Caspase-1、GSDMD-FL、GSDMD-N蛋白表达明显升高(P<0.001)。与模型组比较,益心方各剂量组和曲美他嗪组LVEF、FS显著升高(P<0.05,P<0.001),心肌损伤标志物CK、AST、LDH显著降低(P<0.05,P<0.01,P<0.001),心肌细胞横截面积减小(P<0.05,P<0.001),心肌纤维化程度减轻(P<0.001),心肌细胞焦亡指数下降(P<0.05,P<0.001),心肌组织中NLRP3、pro-Caspase-1、GSDMD-FL、GSDMD-N蛋白表达降低(P<0.001)。结论益心方能够减轻大鼠MIRI后心肌损伤和心肌纤维化程度,从而改善大鼠心功能情况,其机制可能与抑制NLRP3/Caspase-1/GSDMD信号通路介导的心肌细胞焦�Objective To investigate the possible mechanism of myocardial protective effect of Yixin formula in rats with myocardial ischemia-reperfusion injury.Methods Sixty-five rats were randomly divided into 10 in the shamoperated group and 55 in the modeling group;the modeling group was constructed by ligating the left anterior descending branch of the coronary artery,and the sham-operated group was only threaded without ligation.The modeling group was randomly divided into model group,Yixin formula low,medium and high dose group and trimetazidine group after membrane creation.The low,medium and high dose groups of Yixin formula were given 6.84 g·kg^(-1),13.68 g·kg^(-1) and 27.36 g·kg^(-1) of Yixin formula and 5.4 mg·kg^(-1) of Trimetazidine respectively,and the sham-operated group and the model group were given equal volume of saline by gavage for 7 days in each group.Echocardiography was used to detect the cardiac function of rats;biochemical methods were used to detect serum creatine kinase(CK),lactate dehydrogenase(LDH)and glutamate transaminase(AST);hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat myocardium;Masson staining was used to observe the degree of myocardial fibrosis;TUNEL staining for myocardial cell apoptosis index;Western blot was performed to detect NLRP3,pro-Caspase-1 and GSDMD-FL、GSDMD-N protein expression in myocardial tissue.Results Compared with the sham-operated group,the model rats showed significantly lower left ventricular ejection fraction(LVEF)and LV short-axis shortening percentage(FS)(P<0.001),significantly higher myocardial injury markers CK,AST,and LDH(P<0.001),pathological findings showed myocardial structural damage,interstitial edema with inflammatory cell infiltration,disorganized myocardial fiber arrangement,larger myocardial cell cross-sectional The pathological results showed that myocardial structure was damaged,interstitial edema with inflammatory cell infiltration,disorganized myocardial fiber arrangement,large crosssectional area of

关 键 词：心肌缺血再灌注损伤 益心方 核苷酸结合寡聚化结构域样受体蛋白3(NLRP3) 半胱氨酸蛋白酶1(Caspase-1) GSDMD 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]