Mechanism of Buyang Huanwu Tang in Intervening Hypoxia Preconditioning Fibroblast and Promoting Mesenchymal Stem Cell Migration

WANG Yi-jing; YUAN Xiao-peng; WU Dan-bin; SUN Ying-xin; FAN Ying-chang; JIANG Xi-juan; LI Hu-hu

doi:10.13422/j.cnki.syfjx.20182331

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Mechanism of Buyang Huanwu Tang in Intervening Hypoxia Preconditioning Fibroblast and Promoting Mesenchymal Stem Cell Migration

更新时间：2024-09-23

- Mechanism of Buyang Huanwu Tang in Intervening Hypoxia Preconditioning Fibroblast and Promoting Mesenchymal Stem Cell Migration
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 23, Pages: 112-118(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20182331
  CLC： R285.5
- Published：2018
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WANG Yi-jing, YUAN Xiao-peng, WU Dan-bin, et al. Mechanism of Buyang Huanwu Tang in Intervening Hypoxia Preconditioning Fibroblast and Promoting Mesenchymal Stem Cell Migration[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 112-118.
DOI：

WANG Yi-jing, YUAN Xiao-peng, WU Dan-bin, et al. Mechanism of Buyang Huanwu Tang in Intervening Hypoxia Preconditioning Fibroblast and Promoting Mesenchymal Stem Cell Migration[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 112-118. DOI： 10.13422/j.cnki.syfjx.20182331.

摘要

目的：探讨补阳还五汤干预缺氧预适应心脏成纤维细胞促间充质干细胞迁移的相关机制。方法：以6.5，13，26 g·kg-1补阳还五汤和等量蒸馏水连续灌胃SD大鼠7 d后，制备补阳还五汤含药血清及空白血清。0.56，1.12，2.25 g·mL-1含药血清干预缺氧预适应的心脏成纤维细胞（cardiac fibroblasts，CFs）24 h后，酶联免疫吸附测定（ELISA），实时荧光定量聚合酶链式反应（Real-time PCR），蛋白免疫印迹法（Western blot）检测CFs培养液中缺氧诱导因子-1α（hypoxia-inducible factor-1α，HIF-1α），单核细胞趋化因子-1（monocyte chemotactic protein-1，MCP-1）及肝细胞生长因子（hepatocyte growth factor，HGF）的含量，mRNA及蛋白表达情况，并将补阳还五汤干预缺氧预适应CFs的培养液干预间充质干细胞（mesenchymal stem cells，MSC）后，检测MSC的细胞迁移能力。结果：与常氧组比较，缺氧可以显著促进CFs分泌HIF-1α，MCP-1及HGF（P<0.01），同时给予缺氧预处理的CFs培养液干预MSC后，HIF-1α，MCP-1及HGF的水平亦显著升高（P<0.01），当给予不同剂量的补阳还五汤含药血清干预后，与缺氧组比较，它们的表达进一步升高（P<0.05，P<0.01）；而给予HIF-1α抑制剂YC-1处理后，补阳还五汤各组可升高MCP-1，HGF的表达（P<0.05）。结论：补阳还五汤协同缺氧预适应CFs旁分泌效应能够促进MSC迁移，其可能机制是通过上调HIF-1α的表达，提高迁移因子MCP-1，HGF的表达而实现的。

Abstract

Objective:To explore the mechanism of Buyang Huanwu Tang in intervening hypoxia preconditioned cardiac fibroblasts and promoting mesenchymal stem cell migration. Method:The 6.5

26 g·kg-1 Buyang Huanwu Tang and the same volume of distilled water were used to continuously administer SD(Sprague Dawley)rats for 7 days. Then

serum containing Buyang Huanwu Tang and blank serum were prepared. After different doses of serum containing Buyang Huanwu Tang and intervening hypoxia preconditioned cardiac fibroblasts(CFs)for 24 h

enzyme-linked immunosorbent assay(ELISA)

real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the protein levels of hypoxia-inducible factor-1α(HIF-1α)

monocyte chemotactic protein-1(MCP-1)and hepatocyte growth factor (HGF) in CF culture medium. After culture medium intervened the mesenchymal stem cells (MSC)

the MSC were tested for cell migration. Result:Compared with normoxic group

hypoxia could significantly promote the secretion of HIF-1α

MCP-1 and HGF of CFs (P<0.01). At the same time

the levels of HIF-1α

MCP-1 and HGF were significantly increased after intervention of MSC with hypoxia-pretreated CFs (P<0.01). When different doses of serum containing Buyang Huanwu Tang were given

the expressions were further increased compared with the hypoxic group (P<0.05

P<0.01). After HIF-1α inhibitor YC-1 was given

the expressions of MCP-1 and HGF were increased in Buyang Huanwu Tang group(P<0.05). Conclusion:Buyang Huanwu Tang combined with hypoxia preconditioning CFs paracrine effect can promote the migration of MSC

which may be achieved by up-regulating the expression of HIF-1α and increasing the expression of the migration factors MCP-1 and HGF.

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