Effect of Combined Administration of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma Extract on Expressions of VEGF, KDR, AngⅠ and eNOS in Rat Bone Marrow-Derived EPCs

ZHANG Li-fang; LIU Nuan; YANG Lei; WANG Qian; MAO Bing-yu

doi:10.13422/j.cnki.syfjx.20180832

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Effect of Combined Administration of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma Extract on Expressions of VEGF, KDR, AngⅠ and eNOS in Rat Bone Marrow-Derived EPCs

更新时间：2024-01-04

- Effect of Combined Administration of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma Extract on Expressions of VEGF, KDR, AngⅠ and eNOS in Rat Bone Marrow-Derived EPCs
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 10, Pages: 150-155(2018)
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- DOI：10.13422/j.cnki.syfjx.20180832
  CLC： R285.5
- Published：2018
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ZHANG Li-fang, LIU Nuan, YANG Lei, et al. Effect of Combined Administration of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma Extract on Expressions of VEGF, KDR, AngⅠ and eNOS in Rat Bone Marrow-Derived EPCs[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 150-155.
DOI：

ZHANG Li-fang, LIU Nuan, YANG Lei, et al. Effect of Combined Administration of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma Extract on Expressions of VEGF, KDR, AngⅠ and eNOS in Rat Bone Marrow-Derived EPCs[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 150-155. DOI： 10.13422/j.cnki.syfjx.20180832.

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（angiopoietin Ⅰ，AngⅠ）及内皮型一氧化氮合酶（endothelial nitricoxide sythase，eNOS）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eNOS mRNA的表达水平；用蛋白免疫印迹法（Western blot）检测EPCs中VEGF，KDR，AngⅠ和eNOS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eNOS 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eNOS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。

Abstract

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

AngⅠ and eNOS mRNA expressions in EPCs of each treatment group. Western blot was used to detect VEGF

KDR

AngⅠ 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

AngⅠ 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.

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