芳香新塔花总黄酮调控PI3K/Akt/mTOR通路改善动脉粥样硬化的作用机制研究  

Mechanism of total flavonoids of Ziziphora clinopodioides in improving atherosclerosis by regulating PI3K/Akt/mTOR pathway

作　　者：马晓宇 钊浩然 乔会林 曾佑成 张选明[2,1] MA Xiao-yu;ZHAO Hao-ran;QIAO Hui-lin;ZENG You-cheng;ZHANG Xuan-ming(School of Pharmacy,Shihezi University,Shihezi 832000,China;the First Affiliated Hospital of School of Medicine of Shihezi University,Shihezi 832000,China;School of Medicine,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学药学院,新疆石河子832000 [2]石河子大学医学院第一附属医院,新疆石河子832000 [3]石河子大学医学院,新疆石河子832000

出　　处：《中国中药杂志》2023年第2期465-471,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81960853);国家中医药管理局青年岐黄学者支持项目(国中医药人教函(2020)218)。

摘　　要：观察芳香新塔花总黄酮对载脂蛋白E基因敲除(ApoE^(-/-))小鼠自噬和磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的调控,探讨芳香新塔花总黄酮抗动脉粥样硬化(atherosclerosis,AS)的作用机制。ApoE^(-/-)小鼠喂食高脂饲料8周,建立体内AS实验模型,随机分为5组(模型组、阳性对照组及芳香新塔花总黄酮低、中、高剂量组),普通饲料喂食C57BL/6J小鼠设为空白组。灌胃给药12周后取出血清和主动脉,全自动生化分析仪检测小鼠血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平;酶联免疫吸附实验(ELISA)检测小鼠血清中细胞间黏附因子-1(ICAM-1)、血管间黏附因子-1(VCAM-1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)的表达水平;油红O染色法观察小鼠主动脉斑块面积;苏木精-伊红(HE)染色法观察小鼠主动脉斑块及病理变化;免疫荧光法检测主动脉中P62、LC3的表达;蛋白免疫印迹法(Western blot)检测小鼠主动脉中LC3Ⅱ/Ⅰ、Beclin-1、P62、p-PI3K、p-Akt、p-mTOR蛋白表达。结果显示,与空白组相比,模型组小鼠血清中TC、TG、LDL-C、ICAM-1、VCAM-1、MMP-2、MMP-9含量明显升高(P<0.01或P<0.05),HDL-C含量明显降低(P<0.05),主动脉内斑块面积显著增加(P<0.01),主动脉中LC3的表达量明显下调,P62的表达量明显上升(P<0.01或P<0.05),主动脉裂解液中LC3Ⅱ/Ⅰ、Beclin-1的表达量明显下调,p-PI3K、p-Akt、p-mTOR、P62的表达明显上升(P<0.01);芳香新塔花总黄酮中、高剂量组较模型组显示均显著降低血清中TC、TG、LDL-C、ICAM-1、VCAM-1、MMP-2、MMP-9水平(P<0.01或P<0.05),增加HDL-C含量(P<0.01或P<0.05);主动脉斑块面积显著减小(P<0.01),且泡沫细胞的含量减少,管腔的狭窄程度减弱;芳香新塔花总黄酮各剂量组较模型组显示,明显增加主动脉中LC3和裂解液中LC3Ⅱ/Ⅰ、Beclin-1表�The present study observed the regulatory effect of total flavonoids of Ziziphora clinopodioides on autophagy and the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathways in ApoE^(-/-)mice and explored the mechanism of total flavonoids of Z.clinopodioides against atherosclerosis(AS).ApoE^(-/-)mice were fed on a high-fat diet for eight weeks to induce an AS model.The model mice were randomly divided into a model group,a positive control group,and low-,medium-and high-dose groups of total flavonoids of Z.clinopodioides,while C57BL/6J mice fed on a common diet were assigned to the blank group.The serum and aorta samples were collected after intragastric administration for 12 weeks,and the serum levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein-cholesterol(LDL-C),and high density lipoprotein-cholesterol(HDL-C)were detected by an automatic biochemical analyzer.The serum expression levels of intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),matrix metalloproteinase-2(MMP-2),and matrix metalloprotei-nase-9(MMP-9)were detected by enzyme-linked immunosorbent assay(ELISA).Oil red O staining was used to observe the aortic plaque area in mice.Hematoxylin-eosin(HE)staining was used to observe the aortic plaque and pathological changes in mice.The expression of P62 and LC3 in the aorta was detected by the immunofluorescence method.The protein expression of LC3Ⅱ/Ⅰ,Beclin-1,P62,p-PI3K,p-Akt,and p-mTOR in the aorta of mice was detected by Western blot.The results showed that compared with the blank group,the serum levels of TC,TG,LDL-C,ICAM-1,VCAM-1,MMP-2 and MMP-9 in the model group were significantly increased(P<0.01 or P<0.05),the content of HDL-C was decreased(P<0.05),intra-aortic plaque area was enlarged(P<0.01),the expression of LC3 in the aorta was significantly down-regulated,P62 expression was up-regulated(P<0.01 or P<0.05),the expressions of LC3Ⅱ/Ⅰand Beclin-1 in the aortic lysate were significantl

关 键 词：芳香新塔花总黄酮 动脉粥样硬化 自噬 

分 类 号：R285.5[医药卫生—中药学]