基于PI3K/Akt信号通路探讨龙牙楤木总皂苷抗心肌缺血再灌注损伤的机制  

Mechanism of total saponins of Aralia continentalis against myocardial ischemia-reperfusion injury based on PI3K/Akt signal pathway

作　　者：黄炫淳 李慧[2] 鲁卫星 聂静晓 李慧娟 孙丽 HUANG Xuanchun;LI Hui;LU Weixing;NIE Jingxiao;LI Huijuan;SUN Li(Beijing University of Chinese Medicine,Beijing 100029,China;Dongfang Hospital of Beijing University of Chinese Medicine,Beijing 100072,China;The Third Affiliated Hospital of Beijing University of Traditional Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学东方医院,北京100072 [3]北京中医药大学第三临床医学院,北京100029

出　　处：《现代中西医结合杂志》2023年第6期731-737,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金面上项目(81273691);北京市自然科学基金项目(7172124);北京中医药大学新教师启动基金项目(2021-JYB-XJSJJ067)。

摘　　要：目的基于磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路探讨龙牙楤木总皂苷抗心肌缺血再灌注损伤的机制。方法将60只大鼠随机分为假手术组、模型组、龙牙楤木总皂苷组、LY29004组、LY29004+龙牙楤木总皂苷组,每组10只。除假手术组外,其余组利用Langendorff离体心脏灌流装置,通过结扎左冠状动脉前降支30 min、复灌75 min的方法构建离体大鼠心脏再灌注模型。模型组采用K-H液恒速灌流;龙牙楤木总皂苷组和LY29004组分别采用含5 mg/L龙牙楤木总皂苷的K-H液和含15μmol/L LY29004的K-H液恒速灌流;LY29004+龙牙楤木总皂苷组先给予含15μmol/L LY29004的K-H液灌流15 min,再以含5 mg/L龙牙楤木总皂苷的K-H液灌流60 min。采用Western blot法检测心肌组织中PI3K/Akt信号通路相关蛋白[Akt、磷酸化蛋白激酶B(p-Akt)、糖原合酶激酶3β(GSK-3β)、磷酸化糖原合酶激酶3β(p-GSK-3β)]表达情况,免疫组化法检测心肌组织中凋亡相关蛋白[半胱氨酸天门冬氨酸酶3(Caspase-3)、B细胞淋巴瘤基因-2(Bcl-2)、Bcl-2关联蛋白X(Bax)]表达情况,HE染色观察心肌组织病理形态。结果模型组、龙牙楤木总皂苷组、LY29004组、LY29004+龙牙楤木总皂苷组Akt、p-Akt、GSK-3β、p-GSK-3β蛋白相对表达量和Caspase-3、Bax蛋白阳性表达面积均明显高于假手术组(P均<0.05),Bcl-2阳性表达面积均明显低于假手术组(P均<0.05);龙牙楤木总皂苷组Akt、p-Akt、GSK-3β、p-GSK-3β蛋白相对表达量和Bcl-2阳性表达面积均明显高于模型组、LY29004组和LY29004+龙牙楤木总皂苷组(P均<0.05),Caspase-3、Bax蛋白阳性表达面积均明显低于模型组、LY29004组和LY29004+龙牙楤木总皂苷组(P均<0.05)。假手术组心肌细胞排列有序,未见变性、坏死及炎性细胞浸润;模型组大量心肌细胞变性、坏死及炎性细胞浸润;龙牙楤木总皂苷组心肌细胞轻度肿胀,少量细胞变性、坏死,少量炎性细胞浸润;LY29004+龙牙�Objective It is to explore the mechanism of total saponins of Aralia continentalis against myocardial ischemia-reperfusion injury based on phosphatidylinositol-3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods Sixty rats were randomly divided into sham operation group,model group,total saponins of Aralia continentalis group,LY29004 group,LY29004+total saponins of Aralia continentalis group,with 10 rats in each group.Langendorff isolated cardiac perfusion device was used to construct isolated rat heart reperfusion models by ligation of anterior descending branch of left coronary artery for 30min and reperfusion for 75min in all the groups except for the sham operation group.The model group was treated with constant velocity perfusion of K-H liquid,the total saponins of Aralia continentalis group and LY29004 group were treated with constant velocity perfusion of K-H liquid which containing 5 mg/L total saponins of Aralia continentalis and 15μmol/L LY29004,respectively,the LY29004+total saponins of Aralia continentalis group were firstly treated with constant velocity perfusion of K-H liquid containing 15μmol/L LY29004 for 15 minutes,then treated with constant velocity perfusion of K-H liquid containing 5 mg/L total saponins of Aralia continentalis for 60 minutes.The expressions of PI3K/Akt signaling pathway-related proteins[protein kinase B(Akt),phospho protein kinase B(p-AKt),glycogen synthesis kinase 3β(GSK-3β)]and phospho glycogen synthesis kinase 3β(p-GSK-3β)in myocardial tissue were detected by Western blot,the apoptosis related proteins[Cysteinyl aspartate specific proteinase-3(Caspase-3),B-cell lymphoma gene-2(Bcl-2)and Bcl-2 assaciated X protein(Bax)]in myocardial tissue were detected by immunohistochemistry,the pathological morphology of myocardial tissue was observed by HE staining.Results The relative expressions of Akt,p-Akt,GSK-3β,p-GSK-3βand the positive expression areas of Caspase-3 and Bax proteins in the model group,total saponins of Aralia continentalis group,LY29004 group,LY290

关 键 词：龙牙楤木总皂苷 心肌缺血再灌注 磷脂酰肌醇3-激酶 蛋白激酶B 凋亡蛋白 

分 类 号：R-332[医药卫生]