电针预处理调控NLRP3/TRIF/Beclin 1信号通路改善大鼠心肌缺血再灌注损伤  

Electroacupuncture Preconditioning Regulates NLRP3/TRIF/Beclin 1 Signaling Pathway to Improve Myocardial Ischemia-Reperfusion Injury in Rats

作　　者：贺丽丽[1] 姜春宏 刘顺畅 白桦[1] 顾一煌[1] 李开平[1] HE Lili;JIANG Chunhong;LIU Shunchang;BAI Hua;GU Yihuang;LI Kaiping(Nanjing University of Chinese Medicine,Nanjing 210000,Jiangsu,China)

机构地区：[1]南京中医药大学,江苏南京210000

出　　处：《中华中医药学刊》2025年第2期100-104,I0010-I0013,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81974583)。

摘　　要：目的观察电针预处理对心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)大鼠心功能、心肌损伤相关指标及自噬水平的影响,探讨电针预处理改善心肌缺血再灌注损伤的可能机制,进一步发挥中医“治未病”思想指导临床实践的优势。方法将60只雄性SD大鼠随机分为5组:假手术组、模型组、电针组、电针+生理盐水组、电针+激动剂组,每组12只。大鼠结扎左冠状动脉前降支30 min再灌注4 h建立I/R模型。电针组、电针+生理盐水组、电针+激动剂组于“内关”行电针干预,每次20 min,1次/d,连续4 d;电针+激动剂组在每次电针前30 min于腹腔注射NLRP3受体激动剂尼日利亚菌素钠盐Nigericin(1 mg/kg);电针+生理盐水组在每次电针前30 min于腹腔注射生理盐水,剂量同激动剂组。采用超声心动图观察造模后4h后大鼠左心室射血分数(left ventricular ejection fraction,LVEF)、左心室短轴缩短率(left ventricular fractional shortening,LVFS)评估心功能;ELISA法检测血清中炎性因子白细胞介素-6(IL-6)水平;苏木素-伊红(HE)染色法观察大鼠心肌组织形态;实时荧光定量PCR(RT-qPCR)和Western blot法检测NLRP3、Beclin 1、TRIF、Caspase-1、LC3、IL-18 mRNA和蛋白表达。结果与假手术组相比,模型组大鼠LVEF、LVFS值明显降低(P<0.05),心肌纤维排列紊乱、结构模糊、有大片炎性浸润,心肌组织中IL-6水平及NLRP3、TRIF、Beclin 1、LC3 mRNA和蛋白等指标含量均升高(P<0.05);与模型组相比,电针组大鼠LVEF、LVFS明显升高(P<0.05),心肌纤维断裂减轻,少量炎性细胞浸润,心肌组织中IL-6水平、NLRP3 mRNA及蛋白表达均降低(P<0.05),TRIF、Beclin 1、LC3 mRNA和蛋白含量均升高(P<0.05);与电针组相比,电针+生理盐水组LVEF、LVFS及心肌组织中IL-6水平及NLRP3、TRIF、Beclin 1、LC3 mRNA和蛋白含量均无明显变化(P>0.05),而电针+激动剂组大鼠LVEF、LVFS明显降低(P<0.05),心肌结构异常,有明�Objective To observe the effects of electroacupuncture(EA)preconditioning on cardiac function,myocardial injury related indexes and autophagy level in rats with myocardial ischemia-reperfusion injury(MIRI),and to explore the possible mechanism of electroacupuncture preconditioning in improving MIRI.Methods Sixty male SD rats were randomly divided into five groups:sham operation group,model group,EA group,EA+saline group and EA+agonist group,with 12 rats in each group.The I/R model was established by ligation of left anterior descending coronary artery for 30 min and reperfusion for 4 h in rats.The EA group,EA+saline group and EA+agonist group were treated with EA intervention at Neiguan(PC6)for 20 min,once a day for successive 4 days.In EA+agonist group,nigericin(1 mg/kg),an agonist of NLRP3 receptor,was injected intraperitoneally 30 min before each EA.In EA+saline group,saline was injected into abdominal cavity 30 min before each EA,and the dose was the same as that in agonist group.The echocardiography was used to observe left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)of rats 4 hours after modeling to evaluate cardiac function.Serum levels of IL-6 were detected by ELISA.HE staining was used to observe the myocardial tissue morphology of rats.The mRNA and protein expressions of NLRP3,Beclin 1,TRIF,Caspase-1,LC3 and IL-18 were detected by RT-qPCR and Western blot.Results Compared with those of the sham operation group,the values of LVEF and LVFS in the model group were significantly decreased(P<0.05),and the myocardial fibers were disordered,with vague structure and large inflammatory infiltration.The level of IL-6 and the contents of NLRP3,TRIF,Beclin 1,LC3 mRNA and protein in the myocardial tissue were all increased(P<0.05).Compared with those of the model group,the myocardial fiber breakage was reduced and a small amount of inflammatory cells infiltrated.In the EA group,the values of LVEF and LVFS were significantly increased(P<0.05),the level of IL-6 and the expression

关 键 词：电针预处理 心肌缺血再灌注损伤 自噬 NLRP3 Beclin 1 

分 类 号：R285.5[医药卫生—中药学]