摘要
目的:探究黄芪多糖(astragalus polysaccharide,APS)在缺氧条件下促进血管新生的作用机制与靶点。方法:于体外鼠尾胶基质上种植胸主动脉血管段构建新生血管三维培养模型,随机分为对照组、APS25 mg·L^(-1)组,利用倒置显微镜观察新生血管生长情况。体外培养大鼠脑微血管内皮细胞,以氯化钴400μmol·L^(-1)诱导24 h,形成可复制的缺氧模型,随机分为7组(n=6):对照组、缺氧模型组、APS25 mg·L^(-1)组、自噬激动剂雷帕霉素10 mg·L^(-1)组,阳性对照药巴弗洛霉素组20mg·L-1,APS加雷帕霉素组,APS加巴弗洛霉素组。然后利用CCK-8比色法检测各组细胞的增殖活性;蛋白印迹法检测各组细胞自噬相关蛋白Becline-1、LC3B、P62的表达。结果:(1)血管新生实验:与对照组相比,经APS干预新生血管数量明显增多(P<0.05)。(2)细胞实验:与模型组相比,经APS治疗内皮细胞增殖能力提高(P<0.05),Becline-1、LC3B蛋白表达量下调(P<0.01)。结论:在缺氧条件下,黄芪多糖可通过促进血管内皮细胞增殖,促进血管新生,其机制可能与下调内皮细胞Becline-1、LC3B的表达以抑制自噬发生有关。
Objective:Explore the mechanism and target of APS in promoting angiogenesis under hypoxia conditions.Methods:The threedimensional model of vascularization was established by implanting abdominal aorta in Matrigel matrix.After five days,the hypoxia model group and APS 25 mg·L^(-1) group were randomly divided.The growth of new blood vessels was observed by an inverted microscope.Rat brain microvascular endothelial cells were cultured in vitro and induced with cobalt chloride 400μmol·L^(-1) for 24 h to form a replicable hypoxia model,and randomly divided into 7 groups(n=6):control group,hypoxia model group,APS 25 mg·L^(-1) group,autophagy agonist Rapamycin 10 mg·L^(-1) group,positive control drug Bafilomycin A1 group 20 mg·L^(-1),APS plus Rapamycin group,APS plus Bafilomycin A1 group.Then the proliferation activity of each group of cells was detected by the CCK-8 colorimetric method.The expression of autophagy-related proteins Becline-1,LC3B,and p62 in each group was dected by western blot.Results:①Angiogenesis experiment:Compared with the model group,the endothelial cell proliferation ability was improved after APS treatment(P<0.05);②Cell experiment:Compared with the model group,APS treatment significantly increased the proliferation of endothelial cells(P<0.05),down-regulated the expression of Becline-1 and LC3B proteins(P<0.05),and up-regulated the expression of p62 protein(P<0.05).Conclusion:Astragalus polysaccharides can promote angiogenesis by promoting the proliferation of vascular endothelial cells under hypoxia.The mechanism may be related to autophagy inhibition by downregulating the expression of Beclin-1,LC3B,and p62 in endothelial cells.
作者
周严东
高娜
陈叶飞
孙英新
张瑞峰
郑宇佳
李雅文
杨琳
姜希娟
Yandong ZHOU;Na GAO;Yefei CHEN;Yingxin SUN;Ruifeng ZHANG;Yujia ZHENG;Yawen LI;LinYANG;Xijuan JIANG(Tianjin University of Traditional Chinese Medicine Graduate School,Tianjin 300193;Tianjin University of Traditional Chinese Medicine College of Integrated Chinese and Western Medicine,Tianjin 301617;Tianjin University of Traditional Chinese Medicine School of Medical Technology,Tianjin 301617;Shenzhen University Medical School,Shenzhen 518060;Tianjin University of Sport,Tianjin 301617)
出处
《阿尔茨海默病及相关病杂志》
2023年第1期35-42,共8页
Chinese Journal of Alzheimer's Disease and Related Disorders
基金
国家自然科学基金面上项目(82174470)
国家自然科学基金青年项目(81704004)
国家自然科学基金面上项目(81573733)
天津市大学生创新创业基金国家级(201910063024)。
关键词
血管新生
脑微血管内皮细胞
细胞自噬
细胞迁移
黄芪多糖
缺血性脑卒中
Angiogenesis
Cerebral microvascular endothelial cells
Autophagy
Cell migration
Astragalus polysaccharide
Ischemic stroke
