映山红花总黄酮促进大鼠脑血管内皮细胞体外形成血管作用及与VEGFR_(2)和神经源性H_(2)S的关系  

The total flavonoids of Rhodiola japonica promote vascular formation in rat cerebral vascular endothelial cells in vitro and their relationship with VEGFR_(2) and neurogenic H_(2)S

作　　者：仲美静 陈硕 陈志武[1] ZHONG Mei-jing;CHEN Shuo;CHEN Zhi-wu(Dept of Pharmacology,School of Basic Medicine,Anhui Medical University,Hefei 230032,China;Research and Technology Center of Anhui University of Traditional Chinese Medicine,Hefei 230038,China)

机构地区：[1]安徽医科大学基础医学院药理教研室,安徽合肥230032 [2]安徽中医药大学科研技术中心,安徽合肥230038

出　　处：《中国药理学通报》2024年第5期865-872,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 8197141114)。

摘　　要：目的探讨映山红花总黄酮(total flavones of rhododendra,TFR)促大鼠脑血管内皮细胞体外形成血管作用及与VEGFR_(2)和神经源性硫化氢(H_(2)S)的关系。方法采用大鼠脑血管内皮细胞单独培养及和与海马神经元共培养,分别采用不同的实验方法检测细胞增殖、迁移、成管及H_(2)S含量和钙离子荧光强度,包括CCK-8法、细胞划痕法、Transwell法、基质胶成管、H_(2)S试剂盒及钙离子荧光探针法。结果在单独培养的大鼠脑血管内皮细胞上,H_(2)S供体NaHS(200μmol·L^(-1))和TFR(90、270、810 mg·L^(-1))对大鼠脑血管内皮细胞的增殖、迁移、成管及[Ca^(2+)]i荧光强度都有明显的促进作用。而VEGFR_(2)阻断剂SU5416(10μmol·L^(-1))可抑制TFR的促进内皮细胞增殖、迁移和形成血管及[Ca^(2+)]i荧光强度;在与海马神经元共培养的大鼠脑血管内皮细胞上,TFR显著地升高共培养中H_(2)S含量,并被CBS抑制剂AOAA(200μmol·L^(-1))抑制。与此同时,TFR明显地促进共培养中大鼠脑血管内皮细胞的形成血管作用,并可被AOAA和VEGFR_(2)阻断剂SU5416显著地抑制。结论TFR在体外可通过VEGFR_(2)升高[Ca^(2+)]i来促进脑血管内皮细胞形成血管,并可通过诱导神经元中CBS生成H_(2)S作用于大鼠脑血管内皮细胞的VEGFR_(2)来促进血管形成。Aim To explore the effect of total flavonoids of Rhodiola japonica(TFR)on vascular formation in rat cerebral vascular endothelial cells in vitro and its relationship with VEGFR_(2) and neurogenic hydrogen sulfide(H_(2)S).Methods Using rat cerebral vascular endothelial cells cultured separately and co cultured with hippocampal neurons,different experimental methods were used to detect cell proliferation,migration,tube formation,H_(2)S content,and calcium fluorescence intensity,including CCK8 method,cell scratch method,Transwell method,matrix gel tube formation,H_(2)S reagent kit,and calcium fluorescence probe method.Results In isolated cultured rat brain microvascular endothelial cells,H_(2)S donor NaHS(200μmol·L^(-1))and TFR(90,270,810 mg·L^(-1))could significantly promote the proliferation,migration area and the number of migrating cells of primary rat brain microvascular endothelial cells,as well as the formation of blood vessels and[Ca^(2+)]i,while VEGFR_(2) blocker SU5416(10μmol·L^(-1))could inhibit the promotion of endothelial cell proliferation,migration and vascular formation by TFR;TFR significantly increased H_(2)S content in co-cultured rat brain microvascular endothelial cells with hippocampal neurons,and was inhibited by CBS inhibitor AOAA(200μmol·L^(-1)).At the same time,TFR significantly promotes the formation of vascular effects in co-cultured rat cerebral vascular endothelial cells,and could be significantly inhibited by AOAA and VEGFR_(2) blocker SU5416.Conclusions TFR can promote the formation of blood vessels in cerebral vascular endothelial cells by increasing[Ca^(2+)]i of VEGFR_(2) in vitro,and can promote angiogenesis by inducing CBS generation of H_(2)S in neurons and acting on VEGFR_(2) in rat brain microvascular endothelial cells.

关 键 词：硫化氢 海马神经元细胞 大鼠脑血管内皮细胞 增殖 迁移 成管 映山红花总黄酮 

分 类 号：R-322[医药卫生] R322.12R322.81R329.28R392.11R348.1