Effect of Saposhnikoviae Radix-Mume Fructus Drug-containing Serum in Regulating Phenotypic Transformation of Airway Smooth Muscle Cell Proliferation Model

Sheng-zhi LONG; Hai-yan ZHU; Xian-bo WU; Lin CHEN; Yi-qiang XIE; Fan YANG

doi:10.13422/j.cnki.syfjx.20190224

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Effect of Saposhnikoviae Radix-Mume Fructus Drug-containing Serum in Regulating Phenotypic Transformation of Airway Smooth Muscle Cell Proliferation Model

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- Effect of Saposhnikoviae Radix-Mume Fructus Drug-containing Serum in Regulating Phenotypic Transformation of Airway Smooth Muscle Cell Proliferation Model
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 2, Pages: 1-7(2019)
- 作者机构：
  
  1.贵阳中医学院　研究生院，贵阳　 550002；
  2.成都医学院　第一附属医院，成都　 610050；
  3.成都市新都区中医医院，成都　 610050；
  4.海南医学院　中医学院，海口　 571101
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190224
  CLC： R2-0; R22; R285.5
- Received：24 July 2018，
  
  Published Online：02 November 2018，
  
  Published：20 January 2019
- 稿件说明：
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Sheng-zhi LONG, Hai-yan ZHU, Xian-bo WU, et al. Effect of Saposhnikoviae Radix-Mume Fructus Drug-containing Serum in Regulating Phenotypic Transformation of Airway Smooth Muscle Cell Proliferation Model[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(2): 1-7.
DOI：

Sheng-zhi LONG, Hai-yan ZHU, Xian-bo WU, et al. Effect of Saposhnikoviae Radix-Mume Fructus Drug-containing Serum in Regulating Phenotypic Transformation of Airway Smooth Muscle Cell Proliferation Model[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(2): 1-7. DOI： 10.13422/j.cnki.syfjx.20190224.

摘要

目的：

观察防风-乌梅含药血清对气道平滑肌细胞(airway smooth muscle cells，ASMCs)增殖模型表型转化调控的影响，探索防风-乌梅配伍抑制气道重塑的机制，揭示中药配伍机制。

方法：

采用血小板衍生因子(platelet derived growth factor，PDGF)诱导的方式建立ASMCs增殖模型；分别给予大鼠灌胃生理盐水、防风、乌梅、防风-乌梅(生药15.425，15.425，30.85 g·kg

－1

·d

－1

药液)制备药物血清；取4代对数生长期的人支气管平滑肌细胞(human bronchial smooth muscle cell，HBSMC)，将其分为空白组、细胞模型组、正常大鼠血清组、正常大鼠血清细胞模型组、激素干预组、防风血清组、乌梅血清组、防风-乌梅血清组，对细胞给予相应处理，分别采用免疫荧光、蛋白质免疫印迹法(Western blot)检测ASMCs收缩型标志蛋白

-肌动蛋白(

-actin)，合成型标志蛋白骨桥蛋白(osteopontin，OPN)表达，观察表型转化情况；酶联免疫吸附测定(enzyme linked immunosorbent assay，ELISA)检测ASMCs分泌血管内皮生长因子(vascular endothelial growth factor，VEGF)，转化生长因子-

(transforming growth factor-

，TGF-

)，白细胞介素-6(interleukin-6，IL-6)的含量。

结果：

与空白组、正常大鼠血清组比较，模型组、正常大鼠血清细胞模型组

-actin荧光强度与蛋白表达较弱，OPN荧光强度与蛋白表达较强，VEGF，TGF-

，IL-6含量明显提高(

0.05)；与模型组、正常大鼠血清细胞模型组比较，防风-乌梅血清组与激素干预组可显著增强

-actin荧光强度及蛋白表达，降低OPN荧光强度及蛋白表达，降低VEGF，TGF-

，IL-6含量(

0.05)。

结论：

防风-乌梅配伍的抑制气道重塑作用，可能与抑制ASMCs由收缩型向合成型的转化，从而减少VEGF，TGF-

，IL-6等活性物质释放有关。

Abstract

Objective:

To observe the effect of Saposhnikoviae Radix-Mume Fructus containing-serum in regulating the phenotypic transformation of airway smooth muscle cells (ASMCs) proliferation model

in order to explore the mechanism of combined administration of "Saposhnikoviae Radix

Mume Fructus" in inhibiting airway remodeling

and reveal the compatibility mechanism of traditional Chinese medicine.

Method:

The proliferation model of ASMCs was established by platelet derived growth factor (PDGF) induction. The rats were given normal saline

Saposhnikoviae Radix-Mume Fructus

Saposhnikoviae Radix-Mume Fructus(15.425

15.425

30.85 g·kg

-1

·d

-1

) to prepare drug serum respectively. Four generations of logarithmic phase human bronchial smooth muscle cells (HBSMC) were collected and divided into blank control group

cell model group

normal rat serum group and normal rat serum cell model group

hormone intervention group

Saposhnikoviae Radix serum group

Mume Fructus serum group

Saposhnikoviae Radix-Mume Fructus serum group. The cells were given corresponding treatment. Immunofluorescence staining and Western blot were adopted to detect ASMCs deflating marks protein

-actin and osteopontin (OPN) expressions

and phenotypic transformation was observed; the levels of vascular endothelial growth factor(VEGF)

transforming growth factor-

(TGF-

) and interleukin-6(IL-6) secreted by ASMCs were detected by enzyme linked immunosorbent assay (ELISA).

Result:

Compared with blank group and normal rat serum group

the fluorescence intensity and protein expression of

-actin in model group and normal rat serum cell model group were low

whereas the fluorescence intensity and protein expression of OPN were high

and the concentrations of VEGF

TGF-

and IL-6 increased significantly (

0.05). Compared with model group and normal rat serum cell model group

Saposhnikoviae Radix-Mume Fructus serum group and hormone intervention group could significantly enhance the fluorescence intensity and protein expression of alpha-actin

decrease the fluorescence intensity

protein expression of OPN and concentrations of VEGF

TGF-

and IL-6 (

0.05).

Conclusion:

The combined administration of "Saposhnikoviae Radix-Mume Fructus" has an inhibitory effect on airway remodeling

which may be related to the inhibition of the transformation of ASMCs from contractile phenotype to synthetic phenotype

so as to reduce the release of active substances

such as VEGF

TGF-

and IL-6.

关键词

Keywords

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