摘要
目的：探讨黄芪丹参提取物配伍对骨髓源性内皮祖细胞（endothelial progenitor cells,EPCs）中血管内皮生长因子（vascular endothelial growth factor,VEGF）及其受体KDR（kinase insert domain receptor,KDR）,血管生成素Ⅰ（angiopoietinⅠ,AngⅠ）及内皮型一氧化氮合酶（endothelial nitricoxide sythase,e NOS）表达的影响及其可能的作用机制。方法：将离体培养、鉴定后的骨髓源性EPCs,使用黄芪提取物、丹参提取物、黄芪丹参提取物配伍（10,10 mg·L^-1,各5 mg·L^-1）分别处理,并设置空白组,使用逆转录PCR（RT-PCR）及实时荧光定量PCR（Real-time PCR）检测各处理组EPCs中VEGF,KDR,AngⅠ和e NOS mRNA的表达水平;用蛋白免疫印迹法（Western blot）检测EPCs中VEGF,KDR,AngⅠ和e NOS蛋白的表达情况。并使用酶联免疫吸附法（ELISA）检测培养液上清中VEGF的含量。结果：PCR结果显示,与空白组比较,丹参提取物、黄芪提取物组及黄芪丹参提取物配伍组的VEGF,KDR,AngⅠ和e NOS mRNA的表达水平均有显著上调（P〈0.05,P〈0.01）。Western blot结果显示,与空白组比较,丹参提取物、黄芪提取物组VEGF,KDR,AngⅠ和e NOS蛋白表达均有显著上调（P〈0.05,P〈0.01）,而黄芪丹参提取物配伍组中的表达量显著高于其他3组（P〈0.01）,尤其是EPCs细胞培养液上清中的VEGF。ELISA结果显示,黄芪丹参配伍组EPCs中VEGF蛋白含量显著高于其他3组（P〈0.01）。结论：以＂益气化瘀通络生脉＂为治则的黄芪丹参提取物配伍可显著调控EPCs促血管新生作用,其作用可能与上调VEGF密切相关。
Objective：To investigate the effect of the combined administration of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma extract on the expressions of vascular endothelial,growth factor（VEGF）and its receptor kinase insert domain receptor（KDR）,angiopoietin Ⅰ（angiopoietin Ⅰ,AngⅠ） and endothelial nitricoxide synthase（ENOS） in bone marrow-derived endothelial progenitor cells（EPCs）,and its possible mechanism.Method：The bone marrow-derived EPCs after in vitro culture and identification were treated with Astragali Radix extract,Salviae Miltiorrhizae Radix et Rhizoma extract,Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma extract separately（10,10 mg·L^-1,5 mg·L^-1 for each）.The control group was set up.Reverse transcription PCR and quantitative Real-time PCR were used to detect VEGF,KDR,Ang Ⅰ and eNOS mRNA expressions in EPCs of each treatment group.Western blot was used to detect VEGF,KDR,Ang Ⅰ and eNOS protein expressions in EPCs.The content of VEGF in the supernatant of culture fluid was detected by ELISA.Result：PCR results showed that compared with the control group,Salviae Miltiorrhizae Radix et Rhizoma extract group,Astragali Radix extract group and Astragali Radix and combined Salviae Miltiorrhizae Radix et Rhizoma extract group showed significant increases in VEGF,KDR,AngⅠ and eNOS expressions（P〈0.05,P〈0.01）.According to Western blot results,compared with the control group,Salviae Miltiorrhizae Radix et Rhizoma extract group and Astragali Radix extract group showed significant increases in VEGF KDR,Ang Ⅰ and eNOS protein expressions（P〈0.05,P〈0.01）,whereas the expressions of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma extract combination group was significantly higher than that of the other three groups（P〈0.01）,especially VEGF in the culture supernatant of EPCs cells.The results of ELISA showed that the content of VEGF in EPCs of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma extract combination group was significantly higher than that in the other three groups（P〈0.01）.Conclusion：The combination of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma extract can significantly regulate the angiogenic effect of EPCs,which may be closely related to the up-regulation of VEGF.
作者
张瓅方
刘暖
杨雷
王倩
毛秉豫
ZHANG Li-fang;LIU Nuan;YANG Lei;WANG Qian;MAO Bing-yu(Henan Provincial Key Laboratory of ZHANG Zhongjing＇s Formulae and Herbs for lmmunoregulation, Nanyang Institute of Technology, Nanyang 473004, Chin)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第10期150-155,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81473438)
关键词
黄芪
丹参
血管新生
离体骨髓原性内皮祖细胞
血管内皮生长因子
Astragali Radix
Salviae Miltiorrhizae Radix et Rhizoma
angiogenesis
endothelialprogenitor cell
vascular endothelial growth factor
