黄芪协同红花-葶苈子抑制内皮细胞旁分泌改善心肌纤维化机制  

Mechanism of Astragali Radix and Carthami Flos-lepidii Semen in Synergistically Improving Myocardial Fibrosis by Inhibiting the Paracrine Effect of Cardiac Endothelial Cells

作　　者：王咏 王悠草[1] 王丽丽 马度芳 李晓 WANG Yong;WANG Youcao;WANG Lili;MA Dufang;LI Xiao(Department of Cardiology,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Ji'nan 250014,China;Center for Postdoctoral Studies,Shandong University of Traditional Chinese Medicine,Ji'nan 250000,China;Shandong University of Traditional Chinese Medicine,Ji'nan 250000,China)

机构地区：[1]山东中医药大学附属医院心内科,济南250014 [2]山东中医药大学博士后流动站,济南250000 [3]山东中医药大学,济南250000

出　　处：《世界中医药》2025年第3期429-439,共11页World Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82104797);山东省自然科学基金青年项目(ZR2020QH333);山东省济南市科技计划项目(202225004)。

摘　　要：目的:观察黄芪协同红花-葶苈子抑制心肌纤维化的作用及机制。方法:结扎前降支制备心肌梗死后心力衰竭大鼠,红花-葶苈子(C-L)、黄芪(HR)与黄芪-红花-葶苈子(A-C-L)干预,马松染色检测纤维化程度,蛋白质印迹法检测血管性血友病因子(νWF)和α-平滑肌肌动蛋白(α-SMA),蛋白质印迹法及反转录聚合酶链式反应(RT-PCR)检测转化生长因子-β1(TGF-β1)、蜗牛家族转录抑制因子1(Snail)。构建缺氧心脏微血管内皮细胞(HCMEC),C-L、HR及A-C-L含药血清干预,免疫荧光(IF)检测血小板内皮细胞黏附分子(CD31)和α-SMA。A-C-L含药血清联合TGF-β1激活剂(SRI-011381)干预HCMEC,IF观察内皮细胞间充质转化,蛋白质印迹法和RT-PCR检测TGF-β1/Snail表达。提取HCMEC上清液干预心脏成纤维细胞(CF),酶联免疫吸附试验法(ELISA)检测上清液结缔组织生长因子(CTGF),检测CF增殖及胶原合成。结果:A-C-L抑制心力衰竭大鼠心肌纤维化效果最佳。C-L、HR及A-C-L均上调心力衰竭大鼠心肌和缺氧内皮细胞CD31、vWF及下调α-SMA表达,又以A-C-L效果最佳。联合应用SRI-011381后A-C-L抑制内皮间充质转化作用减弱。A-C-L抑制HCMEC中TGF-β1/Snail表达、下调CTGF分泌。提取A-C-L干预的HCMEC上清液培养CF,其CF活化和胶原合成低于HCMEC上清液培养的CF。结论:黄芪协同红花-葶苈子抑制心肌纤维化,其机制,抑制内皮间充质转化,下调内皮细胞中TGF-β1/Snail,以旁分泌方式抑制成纤维细胞增殖和胶原合成。Objective:To investigate the inhibitory effect and mechanism of Astragali Radix(AR)in combination with Carthami Flos-Lepidii Semen(C-L)on myocardial fibrosis.Methods:A rat model of heart failure following myocardial infarction was established by ligation of the left anterior descending coronary artery.The rats were treated with C-L,AR,or Astragali Radix-Carthami Flos-Lepidii Semen(A-C-L).Myocardial fibrosis was assessed using Masson's staining.The expression levels of von Willebrand factor(vWF)and α-smooth muscle actin(α-SMA)were detected by Western blot(WB),while the expression of transforming growth factor-β1(TGF-β1)and Snail family transcriptional repressor 1(Snail)was analyzed using WB and reverse transcription-polymerase chain reaction(RT-PCR).A hypoxic cardiac microvascular endothelial cell(HCMEC)model was constructed,and HCMECs were treated with drug-containing serum from C-L,AR,or A-C-L.The expression of platelet endothelial cell adhesion molecule-1(CD31)andα-SMA was detected by immunofluorescence(IF).To further explore the mechanism of A-C-L in inhibiting endothelial-mesenchymal transition(EMT),HCMECs were co-treated with A-C-L drug-containing serum and the TGF-β1 activator SRI-011381.EMT was assessed via IF,and TGF-β1/Snail expression was detected by WB and RT-PCR.To investigate the paracrine effect,the supernatant from HCMECs was collected and used to treat cardiac fibroblasts(CFs).The levels of connective tissue growth factor(CTGF)in the supernatant were measured by enzyme-linked immunosorbent assay(ELISA),while CF proliferation and collagen synthesis were assessed.Results:A-C-L exhibited the most significant inhibitory effect on myocardial fibrosis in rats with heart failure.C-L,AR,and A-C-L upregulated CD31 and vWF expression while downregulatingα-SMA in both myocardial tissue and hypoxic endothelial cells,with A-C-L demonstrating the strongest effect.After co-treatment with SRI-011381,the inhibitory effect of A-C-L on EMT was weakened.A-C-L inhibited the expression of the TGF-β1/Snail path

关 键 词：心肌纤维化 内皮细胞间充质转化 旁分泌 黄芪 红花-葶苈子药对 转化生长因子-β1/Snail通路 协同 成纤维细胞 心脏微血管内皮细胞 

分 类 号：R285.5[医药卫生—中药学]