中药药理与临床2025，Vol.41Issue(11)：9-15,7.
补阳还五汤配方颗粒调节铜稳态促进缺血组织血管新生的作用机制
Mechanism of Buyang Huanwu Decoction Formula Granules in Promoting Ischemic Tissue Angiogenesis by Regulating Copper Homeostasis
摘要
Abstract
Objective:To explore the role and mechanism of copper in promoting angiogenesis with Buyang Huanwu(补阳还五)Decoction in the angiogenesis of mice after hindlimb ischemia.Methods:Sixty C57BL/6 mice aged 10-12 weeks were adaptively fed for a week and then ran-domly divided into a blank control group,a model control group,a beraprost sodium group(positive control,18 μg/kg/d),and Buyang Huan-wu decoction groups(10,20 and 40 g/kg),with 10 mice in each group.Except for the blank control group,the hindlimb ischemia model was established by left femoral artery ligation,then mice were administered with corresponding drugs or purified water by gavage for 14 days.The laser speckle flow imager was employed to observe blood flow perfusion in the hind limbs.The CD31 antibody was used to stain capillaries in gastrocnemius muscle frozen tissue sections.The copper(Cu)colorimetric assay kit was utilized to detect copper concentration in the gastroc-nemius muscle.Human umbilical vein endothelial cells(HUVECs)were cultured under hypoxia.The following aspects were observed:the effects of Buyang Huanwu Decoction-containing serum and the copper ion chelating agent TTM on endothelial cell proliferation,migration and tube formation,the vascular endothelial growth factor A(VEGFA)expression,and the copper concentration.Chromatin immunoprecipitation(ChIP)and luciferase reporter gene analysis were used to investigate the effects of Buyang Huanwu Decoction-containing serum and TTM on Vegfa gene transcription.Results:Compared with the blank control group,the angiogenesis after limb ischemia was increased in the model control group(P<0.05),while the copper concentration was significantly decreased in ischemic gastrocnemius muscle(P<0.01).Compared with the model control group,the angiogenesis after limb ischemia was increased in each group administered with drugs.Specifically,the best effect occurred in the group given the Decoction at 20g/kg(P<0.01),while the copper concentration of ischemia gastrocnemius muscle was significantly increased in the same group(P<0.01).As for serum,compared with the normal group,the copper concentration,proliferation,migration,and tube formation were significantly increased(P<0.01),while VEGFA expression was raised(P<0.05)in the HUVECs treated with Buyang Huanwu Decoction-containing serum under hypoxia.In contrast,TMM inhibited the above effects(P<0.01).The ChIP and lu-ciferase reporter gene showed that Buyang Huanwu Decoction-containing serum promoted HIF-1α binding activity and HIF-1α mediated Vegfa gene transcription(P<0.01).However,TTM blocked this effect(P<0.01).Conclusion:Buyang Huanwu Decoction increases copper ions in endothelial cells to regulate copper homeostasis,enhances HIF-1α mediated Vegfa transcriptional activity,and promotes endothelial cell pro-liferation,migration,and tube formation,thereby facilitating angiogenesis and restoring blood perfusion in ischemic tissues.关键词
补阳还五汤/缺血/铜稳态/血管新生/血管内皮生长因子AKey words
Buyang Huanwu Decoction/Ischemia/Copper homeostasis/Angiogenesis/Vascular endothelial growth factor A(VEGFA)引用本文复制引用
雷蕴,马涛,万良硕,王世革,张翠娟,程方圆,郭文静,窦永青..补阳还五汤配方颗粒调节铜稳态促进缺血组织血管新生的作用机制[J].中药药理与临床,2025,41(11):9-15,7.基金项目
河北省医学科学研究课题计划项目(编号:20220183) （编号:20220183）
国家自然科学基金项目(编号:82474514) （编号:82474514）
河北省自然科学基金项目(编号:H2021423047) （编号:H2021423047）
河北省中医药类科学研究课题计划项目(编号:2024093) （编号:2024093）
河北中医药大学燕赵医学研究项目(编号:Y2222023010). （编号:Y2222023010）
