千层纸素A动员内皮祖细胞促进血管新生作用机制研究  

Effect of oroxylin A in promoting angiogenesis through mobilizing endothelial progenitor cells:a study on mechanism

作　　者：冷雨泽 张璐莎 孙伟 李梦瑶 薛岳进 陈璐[2] 王虹 Leng Yuze;Zhang Lusha;Sun Wei;Li Mengyao;Xue Yuejin;Chen Lu;Wang Hong(School of Integrative Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;不详)

机构地区：[1]天津中医药大学中西医结合学院,天津301617 [2]方剂学教育部重点实验室、天津市中药药理学重点实验室、天津中医药大学中医药研究院,天津301617

出　　处：《中国循证心血管医学杂志》2022年第11期1329-1333,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：国家国际科技合作专项(2015DFA30430)。

摘　　要：目的探讨千层纸素A通过动员内皮祖细胞发挥促进血管新生作用的机制。方法将30只SPF级健康ICR小鼠制备单侧后下肢缺血模型,按照随机双盲的方法,将后下肢缺血模型造模成功的小鼠分成三组:模型组、千层纸素A组(Oroxylin A)和辛伐他汀组(Sim),每组各10只。采用激光多普勒点扫描成像系统分别检测小鼠术前、术后和给药3、7、14、28d血流量,采用HE染色观察术后28 d缺血腓肠肌组织形态变化;采用流式细胞术检测给药1、3、7d的外周血中内皮祖细胞数量;蛋白芯片检测术后3、7、14d缺血腓肠肌组织CXC型趋化因子配体16(CXCLl6)以及单核细胞趋化性蛋白1(MCP-1)的表达;Western blot检测后下肢缺血小鼠给药3、7、14d基质金属蛋白酶3(MMP3)、基质金属蛋白酶9(MMP9)的表达;采用ELISA检测血浆中肝细胞生长因子(HGF)的表达。结果在后下肢缺血模型小鼠中,千层纸素A组和辛伐他汀组较模型组相比,外周血中GFP+/Sca-1+/Flk-1+内皮祖细胞的数量显著升高,且千层纸素A组小鼠腓肠肌病理形态较模型组明显改善。与模型组比较,给药后3d,千层纸素A组小鼠血浆中HGF的表达显著升高,给药后7d,缺血组织中MMP3、MMP9的蛋白表达量均显著升高。同时EPCs的细胞因子CXCL16及MCP-1的蛋白表达在给药后第7d显著升高。结论千层纸素A通过促进内皮祖细胞介导血管新生,从而发挥促缺血组织血流恢复及损伤修复作用。Objective To study the mechanism of oroxylin A in promoting angiogenesis through mobilizing endothelial progenitor cells(EPCs).Methods SPF-level healthy ICR mice(n=30)were prepared for establishing ischemia model of unilateral posterior leg.The successfully modeling mice were divided,according to randomized double-blinded method,into model group,oroxylin A group and simvatatin group(each n=10).The blood flow volume was detected by using laser Doppler point scanning imaging system before and after the operation,and after medication for 3d,7d,14d and 28d.The morphological changes of ischemic gastrocnemius were observed by using HE staining after the operation for 28d.The number of peripheral blood EPCs was detected by using flow cytometry(FCM)after the medication for 1d,3d and 7d.The expressions of chemokine ligand 16(CXCL16)and monocyte chemoattractant protein-1(MCP-1)in ischemic gastrocnemius were detected by using protein array technique after the operation for 3d,7d and 14d.The expressions of matrix metalloproteinase 3(MMP3)and matrix metalloproteinase 9(MMP9)were detected by using Western blotting assay.The expression of plasma hepatocyte growth factor(HGF)was detected by using ELISA.Results The number of peripheral blood GFP+/Sca-1+/Flk-1+EPCs increased significantly in oroxylin A group and simvatatin group compared with model group,and morphological changes of ischemic gastrocnemius were significantly relieved in oroxylin A group.Compared with model group,the expression of plasma HGF increased significantly after medication for 3d,the protein expressions of MMP3 and MMP9 in ischemic tissue increased significantly after medication for 7d,and meanwhile the protein expressions of CXCL16 and MCP-1 in EPCs increased significantly after medication for 7d in oroxylin A group.Conclusion Oroxylin A can improve angiogenesis through mobilizing EPCs to play the roles of promoting blood flow recovery and damage repair in ischemic tissue.

关 键 词：千层纸素A 内皮祖细胞 血管新生 组织缺血 小鼠 

分 类 号：R541[医药卫生—心血管疾病]