Effects of Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume） Drug-containing Serum on Plateletderived Growth Factor-induced Migration of Airway Smooth Muscle Cells

LONG Shengzhi; ZHU Haiyan; WU Xianbo; CHEN Lin; XIE Yiqiang; YANG Fan; HE Qiansong

doi:10.13288/j.11-2166/r.2019.12.015

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Effects of Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume） Drug-containing Serum on Plateletderived Growth Factor-induced Migration of Airway Smooth Muscle Cells

更新时间：2021-11-10

- Effects of Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume） Drug-containing Serum on Plateletderived Growth Factor-induced Migration of Airway Smooth Muscle Cells
- Journal of Traditional Chinese Medicine Vol. 60, Issue 12, Pages: 1062-1066(2019)
- 作者机构：
  
  1. 贵阳中医学院研究生院
  2. 成都医学院第一附属医院
  3. 成都大学附属医院
  4. 海南医学院中医学院
  5. 成都军区八一骨科医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2019.12.015
  CLC： R285.5
- Published：2019
- 稿件说明：
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[1]龙声志,朱海燕,吴贤波,陈林,谢毅强,杨帆,贺前松.防风乌梅含药血清对血小板衍生生长因子诱导气道平滑肌细胞迁移的影响[J].中医杂志,2019,60(12):1062-1066.

LONG Shengzhi, ZHU Haiyan, WU Xianbo, et al. Effects of Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume） Drug-containing Serum on Plateletderived Growth Factor-induced Migration of Airway Smooth Muscle Cells[J]. Journal of Traditional Chinese Medicine2019, 60(12): 1062-1066.
[1]龙声志,朱海燕,吴贤波,陈林,谢毅强,杨帆,贺前松.防风乌梅含药血清对血小板衍生生长因子诱导气道平滑肌细胞迁移的影响[J].中医杂志,2019,60(12):1062-1066. DOI： 10.13288/j.11-2166/r.2019.12.015.

LONG Shengzhi, ZHU Haiyan, WU Xianbo, et al. Effects of Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume） Drug-containing Serum on Plateletderived Growth Factor-induced Migration of Airway Smooth Muscle Cells[J]. Journal of Traditional Chinese Medicine2019, 60(12): 1062-1066. DOI： 10.13288/j.11-2166/r.2019.12.015.

摘要

目的探讨防风-乌梅配伍对支气管哮喘气道重塑的影响及可能作用机制。方法 20只SD雄性大鼠随机分为正常组、防风组、乌梅组、防风乌梅组（防风∶乌梅=1∶1）

每组5只。防风组、乌梅组、防风乌梅组分别给予相应生药15. 425、15. 425、30. 850 g/（kg·d）药液灌胃

正常组给予等量生理盐水灌胃

各组均每日3次。干预4天后制备含药血清。采用20 ng/ml血小板衍生生长因子诱导的方式促使人支气管平滑肌细胞（HBSMC）迁移模型。实验分为空白对照组、细胞模型组、正常血清组、正常血清细胞模型组、激素干预组、防风血清组、乌梅血清组、防风乌梅血清组

HBSMC以5×104/ml细胞密度接种于培养板

每组8个样本

各组加入10%相应血清干预

37℃、5%CO2培养。培养0、6、24 h时采用细胞划痕实验观察各组细胞迁移率

培养48 h后采用跨膜迁移实验观察跨膜迁移细胞数

ELISA法检测细胞上清液中基质金属蛋白酶9 （MMP-9）、基质金属蛋白酶组织抑制剂1 （TIMP-1）浓度

计算MMP-9/TIMP-1值。结果与空白对照组、正常血清组比较

细胞模型组、正常血清细胞模型组6 h、24 h时细胞迁移率升高

48 h时跨膜迁移细胞数亦升高

细胞上清液中MMP-9浓度、MMP-9/TIMP-1值升高

TIMP-1浓度降低（P <0. 05）。与细胞模型组、正常血清细胞模型组比较

防风乌梅血清组、激素干预组细胞上述各指标均显著改善

并且优于防风血清组和乌梅血清组（P <0. 05）。结论防风-乌梅配伍能显著抑制人支气管平滑肌细胞迁移而影响气道重塑

优于单独使用防风或乌梅

其机制可能与抑制MMP-9/TIMP-1值有关。

Abstract

Objective To investigate the effect of Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume）compatibility on airway remodeling in bronchial asthma and its possible mechanism.Methods A total of 20 male Sprague-Dawley rats were randomly divided into a normal group

a Fangfeng group

a Wumei group and a FangfengWumei group（Radix Saposhnikoviae ∶ Fructus Mume = 1 ∶ 1）

with 5 rats in each group.The Fangfeng group

the Wumei group and the Fangfeng-Wumei group were given the corresponding crude drugs of 15.425

15.425

30.850 g/（kg·d） respectively.The normal group was given the same amount of normal saline.Each group was given 3 times a day.Drug-containing serum was prepared 4 days after the intervention.A human bronchial smooth muscle cell（HBSMC） migration model was induced in a manner induced by 20 ng/ml platelet-derived growth factor.The samples divided into a blank control group

a cell model group

a normal serum group

a normal serum cell model group

a hormone intervention group

a Fangfeng serum group

a Wumei serum group

and a Fangfeng-Wumei serum group.HBSMC was inoculated in 5 × 104/ml cell density on plates

with 8 samples in each group.Each group was added with 10% corresponding serum for intervention

37℃

5% CO2 culture.The cell migration rate of each group was observed by cell scratch test at 0

and 24 h.After 48 h of culture

the number of transmembrane migration cells was observed by transmembrane migration assay.Matrix metalloproteinase 9（MMP-9） and the concentration of matrix metalloproteinase tissue inhibitor 1（TIMP-1） were detected by ELISA.The MMP-9/TIMP-1 value was calculated.Results Compared with the blank control group and the normal serum group

the cell migration rate increased in the cell model group and the normal serum cell model group at 6 h and 24 h

and the number of transmembrane migration cells also increased at 48 h.The MMP-9 concentration and the MMP-9/TIMP-1 value in the cell supernatant increased and the TIMP-1 concentration decreased（P < 0.05）.Compared with the cell model group and the normal serum cell model group

the above indexes of the Fangfeng-Wumei serum group and the hormone intervention group were significantly improved

and were superior to the Fangfeng serum group and the Wumei serum group（P < 0.05）.Conclusion The Fangfeng（Radix Saposhnikoviae）-Wumei（Fructus Mume） compatibility can significantly inhibit the migration of human bronchial smooth muscle cells and affect airway remodeling

which is better than the use of Radix Saposhnikoviae or Fructus Mume alone.The mechanism may be related to the inhibition of MMP-9/TIMP-1.

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School of Acupuncture，Moxibustion，Tuina and Rehabilitation，Hunan University of Chinese Medicine

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