电针预处理基于mTOR/NLRP3信号通路改善心肌缺血再灌注损伤的作用机制  

Mechanism of Electroacupuncture Preconditioning Improving Myocardial Ischemia-Reperfusion Injury Based on mTOR/NLRP3 Signaling Pathway

作　　者：姜春宏 贺丽丽[1] 刘顺畅 白桦[1] 顾一煌[1] 李开平[1] JIANG Chunhong;HE Lili;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3期102-106,I0027-I0029,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81974583)。

摘　　要：目的基于mTOR/NLRP3信号通路探讨电针预处理抗大鼠心肌缺血再灌注损伤的作用机制。方法70只SD大鼠随机分为假手术组、模型组、电针组、电针+生理盐水组,电针+MHY1485(mTOR激动剂)组,各14只。采用超声心动图评估大鼠心脏功能指标心室射血分数(LVEF)和左心室短轴缩短率分数(LVFS),苏木精-伊红(HE)染色法观察心肌组织病理学改变,酶联免疫吸附法(ELISA)检测心肌组织中肿瘤坏死因子-α(TNF-α)水平变化,实时荧光定量PCR(RT-qPCR)检测心肌组织中凋亡相关斑点样蛋白(ASC)、含半胱氨酸的天冬氨酸蛋白水解酶-1(Caspase-1)、白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)的基因表达水平,蛋白质免疫印迹法(Western blot)检测心肌组织中哺乳动物雷帕霉素靶蛋白(mTOR)、NOD样受体家族蛋白3(NLRP3)及核因子-κB(NF-κB)的表达水平。结果与假手术组相比,模型组大鼠FS、EF值下降(P<0.05);心肌组织出现肌纤维断裂、结构模糊、炎性细胞浸润等现象,心肌组织内TNF-α含量升高(P<0.05),Caspase-I、ASC、IL-1β、IL-18的基因相对表达水平(P<0.05)与mTOR、NLRP3、NF-κB的蛋白表达水平均上调(P<0.05);与模型组相比,电针组大鼠FS、EF值上升(P<0.05),心肌组织肌纤维结构改善,炎性细胞浸润减少,心肌组织内的TNF-α含量降低(P<0.05),Caspase-I、ASC、IL-1β、IL-18的基因相对表达水平(P<0.05)与mTOR、NLRP3、NF-κB的蛋白表达水平(P<0.05)均下调;与电针+生理盐水组比,激动剂组大鼠心肌组织损伤加重,心肌组织内TNF-α含量升高(P<0.05),Caspase-I、ASC、IL-1β、IL-18的基因相对表达水平(P<0.05)与mTOR与NLRP3、NF-κB的蛋白表达水平(P<0.05)均上调。结论mTOR激动剂MHY1485可减弱电针对心肌缺血再灌注损伤(MIRI)大鼠心肌组织损伤的保护作用,电针预处理能减轻MIRI大鼠心肌细胞炎性反应,进而改善缺血再灌注引起的心肌组织损伤;因此推断电针预处理改善大鼠MIRI�Objective To investigate the effects of electroacupuncture(EA)preconditioning on myocardial ischemia-reperfu-sion injury(MIRI)and its regulation of the inflammatory response through the mTOR/NLRP3 signaling pathway.Methods Seven-ty SD rats were randomly divided into five groups:sham operation group,model group,EA group,EA+saline group and EA+mTOR agonists(EA+MHY1485)group,with 14 rats in each group.After the intervention,it evaluated cardiac function by meas-uring the left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS).Additionally,it performed he-matoxylin-eosin(HE)staining to observe histopathological changes in the myocardium.The levels of tumor necrosis factor-α(TNF-α)in myocardial tissue were measured using an enzyme-linked immunosorbent assay(ELISA).Furthermore,it as-sessed the gene expression levels of apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate spe-cific proteinase-1(Caspase-1),interleukin-1β(IL-1β)and interleukin-18(IL-18)in cardiac tissue using quantitative polymerase chain reaction(RT-qPCR).Finally,it analyzed the protein expression levels of mammalian target of rapamycin(mTOR),NOD-like receptor thermal protein domain associated protein 3(NLRP3)and nuclear factor kappa-B(NF-κB)by Western blotting.Results Compared to the sham operation group,the rats in the model group showed decreased values of LVFS and LVEF(P<0.05).Myocardial tissue exhibited muscle fiber rupture,blurred structure and a significant infiltration of inflam-matory cells.Additionally,the TNF-αlevel in the myocardial tissue increased(P<0.05).The relative gene expression levels of Caspase-1,ASC,IL-1βand IL-18 were significantly increased(P<0.05),along with increased protein expression levels of mTOR,NLRP3 and NF-κB.In contrast,the rats in the EA group showed increased FS and EF values(P<0.05).The myo-cardial tissue structure improved,inflammatory cell infiltration decreased and the expression of TNF-αin the myocardial tissue decreased(P<0.05).The relative exp

关 键 词：心肌缺血再灌注损伤 电针预处理 炎症 mTOR/NLRP3信号通路 

分 类 号：R245.97[医药卫生—针灸推拿学]