替格瑞洛通过akt/AMPK/eNOS信号通路调节AMI后小鼠血管新生及其机制研究  

Tegretol regulation for post-AMI neovascularisation in mice through the akt/AMPK/eNOS signalling pathway and its mechanism

作　　者：何尤夫 刘微 刘德斌 李玲[1] 向仕菊 姚奇 He Youfu;Liu Wei;Liu Debin;Li Ling;Xiang Shiju;Yao Qi(Department of Cardiovascular Medicine,Guizhou Provincial People’s Hospital,Guiyang 550000,Guizhou,China;Guizhou Cardiovascular Disease Clinical Medical Research Center,Guiyang 550000,Guizhou,China;Department of Cardiovascular Medicine,The Second People’s Hospital of Shantou,Shantou 515000,Guangdong,China;Department of Ultrasound Medicine,Zheng’an County People’s Hospital,Zunyi 563100,Guizhou,China)

机构地区：[1]贵州省人民医院心血管内科,贵州贵阳550000 [2]贵州省心血管疾病临床医学研究中心,贵州贵阳550000 [3]广东省汕头市第二人民医院,广东汕头515000 [4]遵义市正安县人民医院,贵州遵义563100

出　　处：《贵州医药》2024年第4期507-513,519,F0003,共9页Guizhou Medical Journal

基　　金：贵州省科技厅基础研究计划(黔科合基础[2019]1197);贵州省基础研究计划(自然科学类)(黔科合基础-ZK[2023]一般217)。

摘　　要：目的 探究替格瑞洛通过akt/AMPK/eNOS通路调节腺苷浓度,进而促进急性心肌梗死(AMI)后血管新生的作用,并探讨其潜在机制。方法 采用冠脉结扎法建立AMI的小鼠模型,使用替格瑞洛灌胃处理建立实验组,通过akt抑制剂Perifosine、AMPK抑制剂Dorsomorphin和腺苷受体抑制剂MRS1523建立对应蛋白抑制的模型,采用HE染色、Masson染色评估心肌损害情况,采用免疫组化、rt-PCR、Western blot检测VEGF蛋白水平和mRNA水平表达情况,采用酶标法检测AMI小鼠体内腺苷表达情况,随后采用Western blot检测akt/AMPK/eNOS通路蛋白及其磷酸化形式的表达情况。结果 替格瑞洛可明显上调p-akt/akt和p-eNOS/eNOS比值,下降p-AMPK/AMPK比值,并增加AMI后小鼠体内腺苷水平,进而促进心肌梗死后局部心肌细胞内VEGF蛋白水平和mRNA水平表达。结论 替格瑞洛可通过活化akt/AMPK/eNOS系统来调节AMI后小鼠体内腺苷浓度,进而促进局部心肌细胞内VEGF蛋白水平和mRNA水平表达。Objective To investigate the role of tegretol in regulating adenosine concentration through the akt/AMPK/eNOS pathway,thereby promoting post-MI angiogenesis,and explore the underlying mechanisms.Methods A mouse model of AMI was established by coronary ligation.The experimental group was established by Tegretol gavage treatment,and the model corresponding to protein inhibition was established by akt inhibitor perifosine,AMPK inhibitor dorsomorphin and adenosine receptor inhibitor MRS1523.HE staining and Masson’s staining were used to assess myocardial damage.Immunohistochemistry,rt-PCR and Western blot were used to detect the expression of VEGF protein and mRNA levels.Adenosine expression in AMI mice was detected by enzyme labelling.Western blot was then used to detect the expression of akt/AMPK/eNOS pathway proteins and their phosphorylated forms.Results Tegretol could significantly upregulate p-akt/akt and p-eNOS/eNOS ratio,decrease p-AMPK/AMPK ratio and increase adenosine level in vivo in post-AMI mice,which in turn promoted the expression of VEGF protein level and mRNA level in local cardiomyocytes after myocardial infarction.Conclusion Tegretol can regulate adenosine concentration in vivo in post-MI mice through activation of the akt/AMPK/eNOS system,thereby promoting the expression of VEGF protein level and mRNA level in local intracardiac myocytes.

关 键 词：替格瑞洛 急性心肌梗死 腺苷 血管新生 akt/AMPK/eNOS通路 

分 类 号：R542.22[医药卫生—心血管疾病]