天然产物研究与开发 ›› 2024, Vol. 36 ›› Issue (4): 644-652.doi: 10.16333/j.1001-6880.2024.4.011

• 开发研究 • 上一篇    下一篇

补阳还五汤加减通过Nrf2/ARE信号通路防治动脉粥样硬化的机制研究

范增光,袁   野,欧阳效强,赵永法*   

  1. 江西省中医药管理局科技计划(2022A089);江西省自然科学基金(20224BAB216097);江西省卫生健康委科技计划(202210760)
  • 出版日期:2024-04-28 发布日期:2024-04-28
  • 基金资助:
    江西省中医药管理局科技计划(2022A089);江西省自然科学基金(20224BAB216097);江西省卫生健康委科技计划(202210760)

Study on the mechanism of modified Buyang Huanwu Decoction in preventing and treating atherosclerosis through Nrf2/ARE pathway

FAN Zeng-guang,YUAN Ye,OUYANG Xiao-qiang,ZHAO Yong-fa*   

  1. Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,China
  • Online:2024-04-28 Published:2024-04-28

摘要:

基于益心祛瘀化痰法,探讨补阳还五汤加减调控核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)/抗氧化反应元件(antioxidant response element,ARE)信号通路防治动脉粥样硬化的机制。本研究选取ApoE-/-小鼠进行模型复制,模型复制成功后分为模型组、补阳还五汤加减低、中、高剂量组、阿托伐他汀组、C57BL/6J背景的ApoE-/-小鼠空白组。第9周开始灌胃,连续灌胃4周。HE及油红O染色观察小鼠主动脉窦病理学变化;免疫组化法检测高级氧化蛋白产物(advanced oxidative protein product,AOPP)、细胞活性氧(reactive oxygen species,ROS)表达水平;生化分析仪测定血脂表达水平;ELISA法检测丙二醛(malondialdehyde,MDA)、超氧化歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)表达水平;Western blot法检测沉默信息调节因子1(sirtuin1,SIRT1)、Nrf2、血红素氧化酶1(heme oxygenase-1,HO-1)及醌氧化还原酶l(NAD(P)H:quinone oxidoreductase,NQO-1)蛋白表达;RT-PCR法检测HO-1、NQO1基因的mRNA表达。结果显示,与模型组比较,补阳还五汤加减能够改善AS模型小鼠主动脉窦病理学改变(P<0.01);降低主动脉斑块中AOPP、ROS表达水平(P<0.01);升高小鼠血浆中高密度脂蛋白胆固醇表达水平(P<0.01),降低总胆固醇、甘油三酯、低密度脂蛋白胆固醇表达水平(P<0.01);升高SOD、GSH-Px表达水平(P<0.01),降低MDA表达水平(P<0.01);上调小鼠主动脉中SIRT1、Nrf2、HO-1及NQO1蛋白表达水平(P<0.01);并可以上调小鼠主动脉中HO-1、NQO1基因的mRNA表达(P<0.01)。提示补阳还五汤加减通过调控氧化应激损伤发挥抗AS的作用,可能与Nrf2/ARE信号通路活化有关。

关键词: 动脉粥样硬化, 益心祛瘀化痰法, 补阳还五汤, Nrf2/ARE信号通路, 氧化应激损伤

Abstract:

Based on the therapy of benefiting the heart and removing blood stasis and phlegm,this paper discusses the mechanism of modified Buyang Huanwu Decoction (BHD) in preventing atherosclerosis by regulating nuclear factor E2 related factor 2(Nrf2)/antioxidant response element(ARE) signaling pathway.This study selected ApoE-/- mice for model replication.After successful model replication,50 ApoE-/- mice were randomly divided into model group,modified BHD (low,medium and high doses) group,and atorvastatin group,with ten mice in each group;Ten wild-type ApoE-/- mice with C57BL/6J background as a control group.Starting from the 9th week,gavage was administered continuously for four weeks.HE and Oil Red O staining methods were used to observe the pathological and morphological changes of mouse aortic sinuses.Immunohistochemical method was used to detect the expression levels of advanced oxidative protein product(AOPP) and reactive oxygen species(ROS).Biochemical analyzer determination was used to detect the expression levels of blood fat in mouse serum.ELISA method was used to detect the expression levels of oxidative stress related factors such as malondialdehyde(MDA),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in mouse serum.Western blot method was used to detect the expressions of sirtuin1(SIRT1),Nrf2,heme oxygenase-1(HO-1) and NAD(P)H:quinone oxidoreductase(NQO1) proteins in mouse aorta.RT-PCR was used to detect the mRNA expression of HO-1 and NQO1 genes.These results showed that modified BHD could improve the pathological changes of aortic sinus in AS model mice(P<0.01),reduce the lipid content in the aortic sinus of AS model mice(P<0.05),reduce the expression levels of AOPP and ROS proteins in aortic plaques(P<0.01),increase the expression level of how-density lipoprotein cholesterol in mouse serum(P<0.01),and reduce the expression levels of total cholesterol,triglyceride and low-density lipoprotein cholesterol (P<0.01),increase the expression levels of SOD and GSH-Px(P<0.01),and reduce the expression level of MDA(P<0.01),upregulate the expression levels of SIRT1,Nrf2,HO-1 and NQO1 proteins in mouse aorta(P<0.01),and it could also upregulate the mRNA expression levels of HO-1 and NQO1 genes in the mouse aorta(P<0.01).These findings indicated that modified BHD exerts an anti AS effect by regulating oxidative stress damage,and some mechanisms may be related to the activation of Nrf2/ARE signaling pathway.

Key words:

中图分类号:  R285.5 R96