Effects of Shugan Jianpi Huoxue Decoction（疏肝健脾活血方） on Expression of VEGF/VEGFR in Hepatic Stellate Cells/Liver Sinusoidal Endothelial Cell Co-Culture System

LU Jun; ZHOU Ning; HUANG Yan; XIA Xuejiao; ZHANG Junjie

doi:10.13288/j.11-2166/r.2019.03.015

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Effects of Shugan Jianpi Huoxue Decoction（疏肝健脾活血方） on Expression of VEGF/VEGFR in Hepatic Stellate Cells/Liver Sinusoidal Endothelial Cell Co-Culture System

更新时间：2021-11-10

- Effects of Shugan Jianpi Huoxue Decoction（疏肝健脾活血方） on Expression of VEGF/VEGFR in Hepatic Stellate Cells/Liver Sinusoidal Endothelial Cell Co-Culture System
- Journal of Traditional Chinese Medicine Vol. 60, Issue 3, Pages: 242-247(2019)
- 作者机构：
  
  浙江中医药大学基础医学院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2019.03.015
  CLC： R285.5
- Published：2019
- 稿件说明：
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[1]鲁军,周凝,黄棪,夏雪皎,张俊杰.疏肝健脾活血方对肝星状细胞/肝窦内皮细胞共培养体系中VEGF/VEGFR表达的影响[J].中医杂志,2019,60(03):242-247.

LU Jun, ZHOU Ning, HUANG Yan, et al. Effects of Shugan Jianpi Huoxue Decoction（疏肝健脾活血方） on Expression of VEGF/VEGFR in Hepatic Stellate Cells/Liver Sinusoidal Endothelial Cell Co-Culture System[J]. Journal of Traditional Chinese Medicine2019, 60(3): 242-247.
[1]鲁军,周凝,黄棪,夏雪皎,张俊杰.疏肝健脾活血方对肝星状细胞/肝窦内皮细胞共培养体系中VEGF/VEGFR表达的影响[J].中医杂志,2019,60(03):242-247. DOI： 10.13288/j.11-2166/r.2019.03.015.

LU Jun, ZHOU Ning, HUANG Yan, et al. Effects of Shugan Jianpi Huoxue Decoction（疏肝健脾活血方） on Expression of VEGF/VEGFR in Hepatic Stellate Cells/Liver Sinusoidal Endothelial Cell Co-Culture System[J]. Journal of Traditional Chinese Medicine2019, 60(3): 242-247. DOI： 10.13288/j.11-2166/r.2019.03.015.

摘要

目的探讨疏肝健脾活血方逆转肝纤维化进程中肝窦毛细血管化的可能作用机制。方法分离正常大鼠肝星状细胞（HSC）、肝窦内皮细胞（LSEC）和肝纤维化模型大鼠LSEC

利用慢病毒转染得到血管内皮生长因子（VEGF）过表达HSC。实验分为6组

1)正常HSC组:正常大鼠HSC空白血清培养; 2)正常组:正常大鼠HSC及正常LSEC空白血清共培养; 3)模型组:正常大鼠HSC及模型大鼠LSEC空白血清共培养; 4)过表达组:VEGF过表达HSC及模型大鼠LSEC空白血清共培养; 5)模型加中药组:正常大鼠HSC及模型大鼠LSEC 10%含药血清共培养; 6)过表达加中药组:VEGF过表达HSC及模型大鼠LSEC10%含药血清共培养。采用Western Blot法检测各组LSEC中血小板内皮细胞黏附因子CD31及血友病因子（vWF）蛋白表达

HSC中胶原Ⅳ、VEGF及血管内皮生长因子受体2 （VEGFR-2）蛋白表达;采用PCR法检测各组HSC中VEGF、血管内皮生长因子受体1 （VEGFR-1）及VEGFR-2 mRNA表达。结果与正常组比较

模型组及过表达组CD31及vWF蛋白表达升高（P <0. 01）

而模型加中药组较模型组CD31及v WF蛋白表达降低（P <0. 01）;与正常HSC组和正常组比较

模型组及过表达组中胶原Ⅳ、VEGF蛋白表达升高（P <0. 05）

而模型加中药组较模型组VEGF、VEGFR-2蛋白表达降低（P <0. 05）;与正常HSC组和正常组比较

模型组及过表达组VEGF mRNA表达升高（P <0. 01）;模型加中药组VEGF mRNA表达较模型组降低（P <0. 05）。结论疏肝健脾活血方可能通过下调VEGF、VEGFR-2蛋白表达

进而抑制VEGF信号通路

恢复LSEC窗孔结构

从而逆转肝窦毛细血管化。

Abstract

Objective To explore the possible mechanism of Shugan Jianpi Huoxue Decoction（疏肝健脾活血方）in treating liver sinus capillarification in liver fibrosis. Methods The hepatic stellate cells（ HSC） and liver sinusoidal endothelial cell（ LSEC） from normal rats

LSEC from liver fibrosis model rats were separated. The vascular endothelial growth factor（ VEGF） overexpressed HSC was obtained by virus transfection. The cells were divided into six groups: a) normal HSC group

cultured HSC by blank serum; b) normal group: co-culture normal HSC and normal LSEC by blank serum; c) model group: co-culture normal HSC and model LSEC by blank serum; d) overexpression group: co-culture VEGF overexpressed HSC and model LSEC by blank serum; e) model with herbs group:co-culture normal HSC and model LSEC by 10% drug containing serum; f) overexpression with herbs group: coculture VEGF overexpressed HSC and model LSEC by 10% drug containing serum. Western Blot method was used to detect the expression of CD31 and von Willebrand factor（ vWF） in LSEC and the expression of collagen Ⅳ

VEGF and vascular endothelial growth factor receptor 2（ VEGFR-2） in HSC. The expression of VEGF

vascular endothelial growth factor receptor 1（ VEGFR-1） and VEGFR-2 mRNA in HSC was detected by PCR. Results Compared with normal group

the expression of CD31 and vWF in model group and overexpression group was increased（ P < 0. 01）

and the expression of CD31 and v WF in the model with herbs group was decreased when compared with model group（ P < 0. 01）. Compared with normal HSC group and normal group

the expression of collagen Ⅳ and VEGF in model group and overexpression group was increased（ P < 0. 01）

and the expression of VEGF and VEGFR-2 in the model with herbs group was decreased when compared with model group（ P < 0. 05）. Compared with normal HSC group and normal group

the expression of VEGF mRNA in model group and overexpression group was increased（ P < 0. 01）.The expression of VEGF mRNA in model with herbs group was decreased than that in model group（ P < 0. 05）.Conclusion Shugan Jianpi Huoxue Decoction reverse the capillary vascularization of hepatic sinus possibly by downregulating the expression of VEGF and VEGFR-2

inhibiting the signal pathway of VEGF and restoring the window structure of LSEC.

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Related Institution

Zhejiang Chinese Medical University

Graduate School，Beijing University of Chinese Medicine

Guang 'anmen Hospital，China Academy of Chinese Medical Sciences

The Second Affliated Hospital of Guizhou University of Traditional Chinese Medicine

Guizhou University of Traditional Chinese Medicine

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