基础医学与临床 ›› 2022, Vol. 42 ›› Issue (4): 607-615.doi: 10.16352/j.issn.1001-6325.2022.04.022

• 研究论文 • 上一篇    下一篇

虫草素抑制哮喘大鼠支气管上皮细胞间充质转化

汤丽萍1, 闫瑾2, 范芳1, 王浩1, 黄娜1*   

  1. 1.空军军医大学西京医院 儿科, 陕西 西安 710032;
    2.西安医学院 医学技术学院, 陕西 西安 710021
  • 收稿日期:2021-03-17 修回日期:2021-07-02 出版日期:2022-04-05 发布日期:2022-04-01
  • 通讯作者: * 82611@163.com
  • 基金资助:
    陕西省自然科学基础研究计划(2019JM-514)

Cordycepin inhibits bronchial epithelial-mesenchymal transition in asthmatic rats

TANG Li-ping1, YAN Jin2, FAN Fang1, WANG Hao1, HUANG Na1*   

  1. 1 Department of Pediatrics, Xijing Hospital, Air Force Military Medical University, Xi'an 710032;
    2. College of Medical Technology, Xi'an Medical University, Xi'an 710021, China
  • Received:2021-03-17 Revised:2021-07-02 Online:2022-04-05 Published:2022-04-01
  • Contact: * 82611@163.com

摘要: 目的 考察虫草素(Cor)抑制哮喘大鼠上皮细胞间充质转化的作用机制。方法 将大鼠随机分为4组(n=10):对照组、模型(OVA)组、OVA+10 mg/kg Cor组、OVA+50 mg/kg Cor组。治疗7 d后,检测支气管肺泡灌洗液(BALF)中的细胞计数,苏木精-伊红(HE)染色评价气道炎性反应和气道重塑程度,Masson三色染色检测上皮胶原沉积。将人支气管上皮样细胞16HBE分为3组:对照组、TGF-β1组和TGF-β1+Cor组。免疫荧光染色检测细胞中c-Jun的表达。CCK-8法和Transwell小室法检测细胞的增殖和迁移。免疫组织化学染色或Western blot检测E-cadherin、N-cadherin、α-SMA、vimentin、Smad3、p-Smad3、ERK1/2和p-ERK1/2蛋白表达水平。结果 与对照组相比,OVA组的胶原纤维面积和细胞计数显著增加(P<0.05);与OVA组相比,OVA+10 mg/kg Cor组和OVA+50 mg/kg Cor组的胶原纤维面积和细胞计数均减少(P<0.05)。与对照组相比,OVA组的E-cadherin的染色程度和蛋白表达水平均降低,而α-SMA的染色程度升高,N-cadherin、α-SMA和vimentin蛋白表达水平均升高(P<0.05);与OVA组相比,OVA+10 mg/kg Cor组和OVA+50 mg/kg Cor组的E-cadherin的染色程度和蛋白表达水平均升高,而α-SMA的染色程度降低,N-cadherin、α-SMA和vimentin蛋白表达水平均降低(P<0.05)。与对照组相比,TGF-β1组的细胞活力和迁移细胞数均升高(P<0.05);与TGF-β1组相比,TGF-β1+Cor组的细胞活力和迁移细胞数均降低(P<0.05)。与对照组相比,TGF-β1组的E-cadherin蛋白表达水平降低,而N-cadherin、α-SMA、vimentin、p-Smad3和p-ERK1/2蛋白表达水平均升高(P<0.05)。与TGF-β1组相比,TGF-β1+Cor组的E-cadherin蛋白表达水平升高,而N-cadherin、α-SMA、vimentin、p-Smad3和p-ERK1/2蛋白表达水平均降低(P<0.05)。与对照组相比,TGF-β1组的c-Jun的相对荧光强度升高(P<0.05);与TGF-β1组相比,TGF-β1+Cor组的c-Jun的相对荧光强度降低(P<0.05)。结论 Cor通过抑制Smad3和ERK1/2信号通路及c-Jun抑制哮喘中的上皮细胞间充质转化(EMT)。

关键词: 虫草素, 哮喘, 上皮细胞间充质转化, 支气管, 人支气管上皮样细胞

Abstract: Objective To investigate the mechanism of cordycepin (Cor) in the inhibition of epithelial-mesenchymal transition in asthmatic rats. Methods Rats were randomly divided into 4 groups (n=10): control group, ovalbumin (OVA) group, OVA+10 mg/kg Cor group, OVA+50 mg/kg Cor group. After 7 days of treatment, the cell counting in bronchoalveolar lavage fluid (BALF) was measured; hematoxylin-eosin (HE) staining was used to evaluate airway inflammation and airway remodeling. Masson's trichrome staining was used to detect epithelial collagen deposition. Human bronchial epithelioid cells 16HBE were divided into 3 groups: control group, TGF-β1 group and TGF-β1+Cor group. Immuno-fluorescence staining was used to detect the expression of c-Jun in the cells. Cell proliferation and migration were examined by cell counting kit-8 (CCK-8) and Transwell. The protein expression of E-cadherin, N-cadherin, α-SMA, vimentin, Smad3, p-Smad3, ERK1/2 and p-ERK1/2 was detected by immuno-histochemical staining or Western blot. Results With comparison with specific control groups, the following results were recorded. The collagen fiber area and cell count of OVA group increased significantly(P<0.05). The collagen fiber area and cell count of OVA+10 mg/kg Cor group and OVA+50 mg/kg Cor group both decreased (P<0.05). E-cadherin staining degree and protein expression of OVA group decreased, while the staining degree of α-SMA increased and the protein expression of N-cadherin, α-SMA and vimentin increased (P<0.05); E-cadherin staining degree and protein expression of OVA+10 mg/kg Cor group and OVA+50 mg/kg Cor group increased, while the staining degree of α-SMA decreased and the protein expression of N-cadherin, α-SMA and vimentin decreased (P<0.05). Compared with control group, the cell viability and the number of migrating cells in the TGF-β1 group increased (P<0.05); compared with the TGF-β1 group, the cell viability and the number of migrating cells in the TGF-β1+Cor group were both decreased (P<0.05). The expression level of E-cadherin protein in TGF-β1 group decreased, while the protein expression of N-cadherin, α-SMA, vimentin, p-Smad3 and p-ERK1/2 increased (P<0.05). The E-cadherin protein expression level of TGF-β1+Cor group increased, while the N-cadherin, α-SMA, vimentin, p-Smad3 and p-ERK1/2 protein expression levels decreased (P<0.05). The relative fluorescence intensity of c-Jun in TGF-β1 group increased (P<0.05); The relative fluorescence intensity of c-Jun in TGF-β1+Cor group decreased (P<0.05). Conclusions Cordycepin inhibits EMT in asthma by inhibiting Smad3 and ERK1/2 signaling pathway and c-Jun.

Key words: cordycepin, asthma, epithelial-mesenchymal transition, bronchus, human bronchial epithelioid cells

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