通脉养心丸通过SIRT1/FOXO1信号通路改善大鼠心肌缺血再灌注损伤  

Improvement of myocardial ischemia-reperfusion injury in rats through SIRT1/FOXO1 signaling pathway by Tongmai Yangxin Pills

作　　者：于瑞[1,2] 高原[1,2] 李兴渊[1,2] 孙昊堃 朱明军[1,2] 王建茹 YU Rui;GAO Yuan;LI Xingyuan;SUN Haokun;ZHU Mingjun;WANG Jianru(Heart Center,The First Affiliated Hospital of Henan University of CM,Zhengzhou 450000,China;Engineering Research Center for Safety Evaluation and Risk Prevention and Control of Traditional Chinese Medicine in Henan Province,The First Affiliated Hospital of Henan University of CM,Zhengzhou 450000,China;The First Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院心脏中心,郑州450000 [2]河南中医药大学第一附属医院,河南省中药安全评价与风险防控工程研究中心,郑州450000 [3]河南中医药大学第一临床医学院,郑州450000

出　　处：《中华中医药杂志》2024年第12期6755-6761,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河南省科技攻关项目(No.202102310492,No.212102311079,No.242102310527);河南省中医药科学研究专项课题(No.2024ZY3027);河南省青年人才托举项目(No.2024HYTP043);国家自然科学基金重点项目(No.82004311)。

摘　　要：目的:探讨通脉养心丸(TMYXP)对大鼠心肌缺血再灌注损伤(MIRI)的影响及可能的作用机制。方法:SD大鼠随机分为假手术组、模型组、TMYXP低剂量组、TMYXP中剂量组、TMYXP高剂量组和抑制剂组,每组18只。假手术组和模型组给予0.5%羧甲基纤维素钠灌胃;TMYXP低、中、高剂量组分别给予1.0、2.0、4.0 g·kg^(-1)·d^(-1)TMYXP灌胃;抑制剂组给予4.0 g·kg^(-1)·d^(-1)TMYXP灌胃并腹腔注射EX-527(5 mg·kg^(-1)·d^(-1))。连续干预7 d,末次给药后,利用结扎冠状动脉前降支30 min,再灌注2 h,构建大鼠MIRI模型,假手术组操作同模型组,但前降支仅穿线不结扎。TTC染色法评价大鼠心肌梗死面积;TUNEL染色法观察心肌细胞的凋亡;试剂盒检测血清中乳酸脱氢酶(LDH)和肌酸激酶同工酶MB(CK-MB)水平,心肌组织中超氧化物歧化酶(SOD)、丙二醛(MDA)和过氧化氢酶(CAT)含量;沉默信息调节因子1(SIRT1)去乙酰化酶荧光测定试剂盒检测SIRT1活性;Western Blot检测SIRT1/FOXO1信号通路中SIRT1、乙酰化叉头状转录因子1(FOXO1)、锰超氧化物歧化酶(MnSOD)、B细胞淋巴瘤2(BCL-2)、BCL-2相关X(Bax)、Cleaved-Caspase-3蛋白表达水平。结果:与假手术组比较,模型组大鼠心肌梗死面积,LDH、CK-MB和MDA水平,Acetyl-FOXO1、Bax、Cleaved-Caspase3蛋白表达均显著升高(P<0.05);心肌组织存在大量TUNEL阳性细胞;SOD和CAT水平,SIRT1活性,SIRT1、MnSOD、BCL-2蛋白表达均显著降低(P<0.05)。与模型组比较,TMYXP低、中、高剂量组大鼠心肌梗死面积,LDH、CK-MB和MDA水平,Acetyl-FOXO1、Bax、Cleaved-Caspase-3蛋白表达均显著降低(P<0.05);TUNEL阳性细胞数目均减少;SOD和CAT水平,SIRT1活性,SIRT1、MnSOD、BCL-2蛋白表达均显著升高(P<0.05)。与TMYXP高剂量组比较,抑制剂组大鼠心肌梗死面积,LDH、CK-MB和MDA水平,Acetyl-FOXO1、Bax、Cleaved-Caspase3蛋白表达均显著升高(P<0.05);TUNEL阳性细胞数目有所增加;SOD和CAT水平,SIRT1活性,SIRT1、MnSOD�Objective:To explore the effects and possible mechanism of Tongmai Yangxin Pills(TMYXP)on myocardial ischemia-reperfusion injury(MIRI)in rats.Methods:SD rats were randomly divided into sham-operation group,model group,TMYXP low-dose group,TMYXP medium-dose group,TMYXP high-dose group and inhibitor group,18 rats in each group.The sham operation group and model group were given 0.5%carboxymethylcellulose sodium by gavage;the TMYXP low,medium,and high dose groups were given 1.0,2.0 and 4.0 g·kg^(-1)·d^(-1)of TMYXP by gavage,respectively;and the inhibitor group was given 4.0 g·kg^(-1)·d^(-1)of TMYXP by gavage and intraperitoneal injection of EX-527(5 mg·kg^(-1)·d^(-1)).The rat MIRI model was constructed by ligating the anterior descending branch of the coronary artery for 30 min and reperfusion for 2 h after the last administration of the drug for 7 days.The operation of the sham-operated group was the same as that of the model group,but the anterior descending branch was threaded only and not ligated.The area of myocardial infarction in rats was evaluated with TTC staining method;apoptosis of cardiomyocytes was observed with TUNEL staining;and the kit detected the lactate dehydrogenase(LDH)and creatine kinase isoenzyme(CK-MB)in the serum,and superoxide dismutase(SOD),malondialdehyde(MDA)and catalase(CAT)levels in myocardial tissues;silent information regulator 1(SIRT1)deacetylase fluorescence assay kit measured SIRT1 activity;Western Bolt was detected SIRT1,acetylated forkhead-like transcription factor 1(Acetyl-FOXO1),manganese superoxide dismutase(MnSOD),B-cell lymphoma 2(BCL-2),BCL-2-associated X(Bax),and Cleaved-Caspase-3 protein expression levels in the SIRT1/FOXO1 signaling pathway.Results:Compared with the sham-operated group,myocardial infarction area,LDH,CK-MB,and MDA levels,and protein expression of Acetyl-FOXO1,Bax,and Cleaved-Caspase-3 were significantly higher in the model group of rats(P<0.05);a large number of TUNEL-positive cells were present in myocardial tissues;and SOD and CAT levels,SIRT1 ac

关 键 词：心肌缺血再灌注损伤 通脉养心丸 氧化应激 细胞凋亡 沉默信息调节因子1/叉头状转录因子1信号通路 中医药 机制 

分 类 号：R73[医药卫生—肿瘤]