补阳还五汤含药血清对人肺动脉内皮细胞间质转化的影响

渠景连; 杨邯捷; 赵惠亮

doi:10.13422/j.cnki.syfjx.20192036

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补阳还五汤含药血清对人肺动脉内皮细胞间质转化的影响

经典名方 | 更新时间：2021-02-09

- 补阳还五汤含药血清对人肺动脉内皮细胞间质转化的影响
- Effect of Serum Containing Buyang Huanwu Tang on Endothelial-to-mesenchymal Transition of Human Pulmonary Artery Endothelial Cells
- 中国实验方剂学杂志 2019年25卷第19期页码：1-9
- 作者机构：
  
  贵州中医药大学，贵阳　 550025
- 作者简介：
  
  渠景连，博士，讲师，从事温病治则治法的机制研究，E-mail：qqpsy2009@126.com
  赵惠亮，硕士，讲师，从事中医心理学研究，E-mail：413385641@qq.com
- 基金信息：
  
  国家自然科学基金项目(81660789);贵州省科技厅科技计划项目(黔科合基础[2017]1013);贵州省教育厅青年科技人才成长项目(黔教合KY字[2017]177);贵州省中医药管理局中医药、民族医药科学技术研究项目(QZYY-2018-104)
- DOI：10.13422/j.cnki.syfjx.20192036
  中图分类号： R2-0; R285.5; R289; R318.13
- 收稿日期：2019-04-13，
  
  网络出版日期：2019-07-04，
  
  纸质出版日期：2019-10-05
- 稿件说明：
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渠景连, 杨邯捷, 赵惠亮. 补阳还五汤含药血清对人肺动脉内皮细胞间质转化的影响[J]. 中国实验方剂学杂志, 2019,25(19):1-9.

Jing-lian QU, Han-jie YANG, Hui-liang ZHAO. Effect of Serum Containing Buyang Huanwu Tang on Endothelial-to-mesenchymal Transition of Human Pulmonary Artery Endothelial Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(19): 1-9.
渠景连, 杨邯捷, 赵惠亮. 补阳还五汤含药血清对人肺动脉内皮细胞间质转化的影响[J]. 中国实验方剂学杂志, 2019,25(19):1-9. DOI： 10.13422/j.cnki.syfjx.20192036.

Jing-lian QU, Han-jie YANG, Hui-liang ZHAO. Effect of Serum Containing Buyang Huanwu Tang on Endothelial-to-mesenchymal Transition of Human Pulmonary Artery Endothelial Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(19): 1-9. DOI： 10.13422/j.cnki.syfjx.20192036.

摘要

目的：

研究补阳还五汤含药血清对人肺动脉内皮细胞(HPAEC)间质转化(EndMT)的调控作用，并从Jagged1/Notch1信号传导进一步分析其作用机制。

方法：

以53.36 g·kg

－1

·d

－1

剂量灌胃家兔，制备补阳还五汤含药血清，等容积生理盐水灌胃制备空白血清。体外培养HPAEC细胞，转化生长因子-

(TGF-

)诱导建立EndMT模型，设立空白组(10%空白血清)，模型组(10%空白血清＋TGF-

)，补阳还五汤含药血清高剂量组(10%含药血清＋TGF-

)，补阳还五汤含药血清中剂量组(5%含药血清＋5%空白血清＋TGF-

)和补阳还五汤含药血清低剂量组(2.5%含药血清＋7.5%空白血清＋TGF-

)5组。采用噻唑蓝(MTT)比色法检测细胞增殖能力，transwell和划痕实验检测细胞迁移能力，免疫荧光观察内皮细胞标志物血小板内皮细胞黏附分子1(CD31)，血管内皮钙黏蛋白(VE-cadherin)和间质细胞标志物成纤维细胞特异性蛋白1(FSP1)，

-平滑肌肌动蛋白(

-SMA)的表达，蛋白免疫印迹法(Western blot)检测Notch1，Jagged1及CBF1蛋白的表达。

结果：

与空白组比较，模型组HPAEC细胞增殖能力、迁移能力均显著增强(

0.01)，CD31和VE-cadherin表达呈下降趋势，FSP1和

-SMA表达呈上升趋势，Notch1，Jagged1和CBF1表达显著上调(

0.01)。补阳还五汤含药血清干预后，与模型组比较，细胞增殖能力、迁移能力均明显减弱(

0.05，

0.01)，且CD31和VE-cadherin表达呈上升趋势，FSP1和

-SMA表达呈下降趋势，Notch1，Jagged1和CBF1表达明显降低(

0.05，

0.01)，随着含药血清浓度的增加，作用越明显。

结论：

补阳还五汤含药血清可部分抑制人肺动脉内皮细胞发生间质转化，这一过程可能与调控Jagged1/Notch1信号有关。

Abstract

Objective:

To investigate the regulatory effect of serum containing Buyang Huanwu Tang on endothelial-to-mesenchymal transition(EndMT) in human pulmonary artery endothelial cells (HPAEC)

and make further analysis on its mechanism from the perspective of the signal transduction of Jagged1/Notch1.

Method:

Rabbit serum containing Buyang Huanwu Tang was prepared by gavage with dosage of 53.36 g·kg

-1

·d

-1

and blank serum was prepared by gavage with same volume of normal saline. The HPAECs cultured

in vitro

EndMT model was established by the transforming growth factor-

(TGF-

) induced

which were divided into five groups: the control group (10%blank serum)

the model group (10%blank serum+ TGF-

)

the serum containing high-dose Buyang Huanwu Tang group (10%medicated serum + TGF-

)

the medium-dose group (5%medicated serum + 5%blank serum medicated + TGF-

) and the low-dose group(2.5%medicated serum+ 7.5%blank serum+ TGF-

). Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) method. Cell migration was detected by transwell and scratch assay. The endothelial markers platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31)

vascular endothelial cadherin (VE-cadherin) and the mesenchymal markers fibroblast-specific protein 1 (FSP1)

-smooth muscle actin (

-SMA) were observed by immunofluorescence assay. The expression levels of Notch1

Jagged1 and CBF1 were detected by Western blot assay.

Result:

Compared with the control group

the proliferation and migration abilities of the HPAEC cells in model group were enhanced (

0.01)

and the expressions of CD31 and VE-cadherin were decreased

while the expressions of FSP1 and

-SMA were increased. Further study found that the expressions of Notch1

Jagged1 and CBF1 were up-regulated (

0.01). After the intervention of the serum containing Buyang Huanwu Tang

compared with model group

the proliferation and migration abilities of cells were decreased (

0.05

0.01)

and the expressions of CD31 and VE-cadherin were on the rise

while the expressions of FSP1and

-SMA were on the decline. The expressions of Notch1

Jagged1 and CBF1 were also significantly lower than those in model group (

0.05

0.01). With the increase of serum concentration

the effect was more obvious.

Conclusion:

The serum containing Buyang Huanwu Tang can partly inhibit the EndMT in human pulmonary artery endothelial cells

which may be related to the regulation effect of Jagged1/Notch1 signaling.

关键词

Keywords

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