Guben Fangxiao decoction Regulates Expression of CGRP /ILC2 and GABA in Human Bronchial Epithelial Cells Stimulated with IL-4 Combined with IL-13
10.13422/j.cnki.syfjx.20241142
- VernacularTitle:固本防哮饮对IL-4联合IL-13诱导的人支气管上皮细胞中CGRP/ILC2及GABA表达的影响
- Author:
Weiying OU
1
;
Xuejing YUAN
1
;
Jianjian JI
2
Author Information
1. Affiliated Hospital of Nanjing University of Chinese Medicine， Nanjing 210029， China
2. The First Clinical Medical College of Nanjing University of Chinese Medicine， Nanjing 210023， China
- Publication Type:Journal Article
- Keywords:
bronchial asthma;
Guben Fangxiao decoction;
calcitonin gene-related peptide;
γ-aminobutyric acid;
type Ⅱ innate lymphoid cell
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(2):93-100
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo observe the effects of Guben Fangxiao decoction-containing serum on the expression of transcription factors and cytokines associated with calcitonin gene-related peptide （CGRP）， γ-aminobutyric acid （GABA）， and type Ⅱ innate lymphoid cells （ILC2） in human bronchial epithelial cells （BEAS-2B） stimulated with interleukin-4 （IL-4） combined with interleukin-13 （IL-13）， and explore the possible mechanism of Guben Fangxiao decoction-containing serum in alleviating type Ⅱ inflammation of bronchial epithelial cells. MethodsBEAS-2B cells stimulated with IL-4 combined with IL-13 were treated with the sera containing high （20%）， medium （15%）， and low （10%） doses of Guben Fangxiao decoction and the montelukast-containing serum （10%）. Real-time PCR was used to measure the mRNA levels of CGRP， GABA， mucin 5AC （MUC5AC）， thymic stromal lymphopoietin （TSLP）， interleukin （IL）-25， IL-33， IL-5， GATA-binding protein 3 （GATA3）， and B cell lymphoma/leukemia 11B （Bcl-11B） in each group. Western blot was employed to determine the relative protein levels of CGRP and MUC5AC. The immunofluorescence assay was employed to observe the relative expression of CGRP and GABA in the cells. Enzyme-linked immunosorbent assay was used to quantify the protein levels of IL-33， MUC5AC， and IL-5 in the culture and cell supernatants. Periodic acid-Schiff staining was performed to assess mucin secretion. ResultsCompared with the control group， the model group showed up-regulated mRNA levels of CGRP， GABA， MUC5AC， TSLP， IL-25， and IL-33 （P<0.05， P<0.01）. Compared with the normal group， the Guben Fangxiao decoction-containing serum down-regulated the mRNA levels of CGRP， GABA， MUC5AC， IL-5， GATA3， and Bcl-11B （P<0.05， P<0.01） in a dose-dependent manner. Compared with the normal group， the model group showed up-regulated protein levels of CGRP， GABA， MUC5AC， IL-33， and IL-5 （P<0.05， P<0.01）， which were down-regulated by the Guben Fangxiao decoction-containing serum （P<0.05， P<0.01）. Compared with the normal group， the model group showed enhanced average fluorescence intensity of CGRP and GABA （P<0.01）， which was weakened by the Guben Fangxiao decoction-containing serum （P<0.01）. Compared with the normal group， the model group showed increased secretion of mucin， which was reduced by the Guben Fangxiao decoction-containing serum. ConclusionGuben Fangxiao decoction can treat bronchial asthma by alleviating type Ⅱ immune airway inflammation and reducing airway mucus secretion， which is achieved by regulating the expression of CGRP， GABA， MUC5AC， and ILC2-related transcription factors.
