益气养阴化瘀汤调节CB2R/β-arrestin-2通路相关的自噬保护糖尿病心肌病模型小鼠的心功能  

Yiqi Yangyin Huayu Decoction regulates CB2R/β-arrestin-2 pathway-related autophagy to protect cardiac function in a diabetic cardiomyopathy mouse model

作　　者：章睿[1] 郑萤萤 戴圣伟[2] ZHANG Rui;ZHENG Yingying;DAI Shengwei(Department of Rehabilitation Medicine,Zhejiang Hospital,Hangzhou,Zhejiang 310013,China;Department of Emergency Medicine,Lishui Hospital of Traditional Chinese Medicine,Lishui,Zhejiang 323900,China)

机构地区：[1]浙江医院康复医学科,杭州310013 [2]浙江省丽水市中医院急诊科,丽水323900

出　　处：《浙江中西医结合杂志》2025年第4期305-312,共8页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.2021ZB007);浙江省医药卫生科技计划项目(No.2020KY390)。

摘　　要：目的探讨益气养阴化瘀汤对糖尿病心肌病(DCM)模型小鼠心脏功能和心肌组织炎症的影响,并阐明其通过大麻素受体2(CB2R)/抑制蛋白β2(β-arrestin-2)通路调节自噬的机制。方法动物实验:C57BL/6小鼠20只,按随机数字表法分为对照组、模型组、低剂量中药组和高剂量中药组,每组5只。采用高脂饮食和链脲佐菌素诱导小鼠DCM模型。低、高剂量中药组造模成功后分别给予益气养阴化瘀汤14.35、28.70 mg/(kg·d)灌胃,对照组、模型组小鼠予生理盐水2 mL/(kg·d)灌胃,各组均干预6周。检测各组小鼠血糖水平。超声心动图评估各组小鼠心脏功能。苏木精-伊红(HE)染色检测小鼠心肌组织病理变化。酶联免疫吸附试验(ELISA)检测小鼠心肌组织炎症因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平。蛋白质印迹(WB)检测CB2R/β-arrestin-2、腺苷酸激酶(AMPK)/雷帕霉素靶蛋白(mTOR)通路以及自噬相关蛋白CB2R、β-arrestin-2、螯合体1(p62)、微管相关蛋白1轻链3Ⅰ/Ⅱ(LC3Ⅰ/Ⅱ)、B淋巴细胞瘤-2基因同源结构域蛋白(Beclin-1)、磷酸化腺苷酸激酶(p-AMPK)/AMPK和磷酸化雷帕霉素靶蛋白(p-mTOR)/mTOR表达水平。细胞实验:将小鼠原代心肌细胞随机分为对照组、模型组、CB2R过表达组、对照载体组。WB检测各组小鼠心肌细胞CB2R、β-arrestin-2、p62、LC3Ⅰ/Ⅱ、Beclin-1表达水平。结果动物实验:与模型组比较,干预后低、高剂量中药组小鼠血糖[(21.17±2.70)mmol/L、(14.15±1.94)mmol/L比(25.53±3.45)mmol/L,P<0.05或P<0.01]显著降低,超声心动图心脏射血分数(EF)[(80.89±4.45)%、(74.89±4.11)%比(63.31±6.45)%,P<0.01]、缩短分数(FS)[(34.78±4.15)%、(41.51±1.99)%比(25.13±2.16)%,P<0.01]显著升高,左室内径(LVIDd)[(3.75±0.36)mm、(3.55±0.46)mm比(4.35±0.25)mm,P<0.01]显著降低,小鼠心肌组织炎症因子IL-1β[(32.80±1.95)pg/mL、(15.00±2.50)pg/mL比(44.05±7.26)pg/mL,P<0.05]、IObjective To investigate the effects of Yiqi Yangyin Huayu Decoction(YYHD)on cardiac function and myocardial tissue inflammation in a diabetic cardiomyopathy(DCM)mouse model,and to elucidate its mechanism in regulating autophagy through the cannabinoid receptor 2(CB2R)/β-arrestin-2 pathway.Methods Animal experiment:A total of 20 C57BL/6 mice were randomly divided into four groups using a random number table:control group,model group,and low-and high-dose herbal groups,with 5 mice in each group.The DCM model was induced in mice using a high-fat diet and streptozotocin.After successful modeling,the low-and high-dose herbal groups were treated with YYHD by gavage at doses of 14.35 and 28.70 mg/(kg·d),respectively.The control and model groups received an equal volume of physiological saline by gavage.All groups were treated for 6 weeks.Blood glucose levels of the mice in each group were measured.Cardiac function was assessed using echocardiography.Hematoxylin and eosin(HE)staining was performed to detect histopathological changes in myocardial tissue.Enzyme-linked immunosorbent assay was used to measure the levels of pro-inflammatory cytokines interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in myocardial tissues.Western blotting(WB)was used to assess the expression levels of proteins related to the CB2R/β-arrestin-2 pathway,the AMPK/mTOR pathway,and autophagy-related proteins,including CB2R,β-arrestin-2,p62,microtubule-associated protein 1 light chain 3Ⅰ/Ⅱ(LC3Ⅰ/Ⅱ),Beclin-1,p-AMPK/AMPK,and p-mTOR/mTOR.Cell experiment:Primary cardiomyocytes from mice were randomly divided into a control group,model group,CB2R overexpression group,and control vector group.WB was used to measure the expression levels of CB2R,β-arrestin-2,p62,LC3Ⅰ/Ⅱ,and Beclin-1 in cardiomyocytes from each group.Results Animal experiment:Compared to the model group,after treatment,the blood glucose levels in the low-and high-dose herbal groups were significantly lower[(21.17±2.70)mmol/L,(14.15±1.94)mmol/L

关 键 词：小鼠 糖尿病心肌病 益气养阴化瘀汤 自噬 大麻素受体2/抑制蛋白β2 腺苷酸激酶/雷帕霉素靶蛋白 

分 类 号：R285.5[医药卫生—中药学]