Effects of Zhike Pingchuan Formula on airway inflammation and TLR4/TRAF6/NF-κB pathway in bronchial asthma mice

doi:10.11958/20240222

Abstract

Abstract:

Objective To investigate the effect and mechanism of Zhike Pingchuan Formula on reducing airway inflammation in mice with bronchial asthma (BA). Methods A total of 84 BALB/c mice were randomly separated into the control group, the BA group, the low-dose Zhike Pingchuan Formula group, the high-dose Zhike Pingchuan Formula group, the dexamethasone group, the lipopolysaccharide (LPS) group and the high-dose Zhike Pingchuan Formula+LPS group, with 12 mice in each group. The BA model was constructed in all groups of mice except the control group. After successful modeling, mice were treated with drug administration once a day for 3 weeks. Serum immunoglobulin E (IgE) concentration and levels of interleukin-17 (IL-17) and tumor necrosis factor-α (TNF-α) in alveolar lavage fluid were detected by enzyme linked immunosorbent assay (ELISA). Pathological changes in lung tissue was detected by HE staining. The bronchial epithelium goblet cell proliferation and mucus secretion were determined by PAS staining. Flow cytometry was used to detect levels of T helper cell 17 (Th17) and regulatory T (Treg) cell in mouse spleen tissue. Western blot assay was used to detect toll-like receptor 4 (TLR4)/tumor necrosis factor receptor-associated factor 6 (TRAF6)/nuclear factor kappa-B (NF-κB) pathway-related protein expression in lung tissue. Results Compared with the control group, damage of lung tissue was serious, the proliferation of bronchial epithelial goblet cells and the secretion of mucus increased, IgE concentration, TNF-α, IL-17 levels, Th17 cell proportion, Th17/Treg ratio, and TLR4, TRAF6, p-NF-κB p65 protein expression increased, while Treg cell proportion decreased in the BA group (P<0.05). Compared with the BA group, the bronchial epithelial cupular cell proliferation and mucus secretion were reduced in the low-dose Zhike Pingchuan Formula group, the high-dose Zhike Pingchuan Formula group and the dexamethasone group, the lung tissue damage was improved, the concentration of IgE, levels of TNF-α, IL-17, the proportion of Th17 cells, the ratio of Th17/Treg and expression of protein levels of TLR4, TRAF6 and p-NF-κB p65 were reduced, and the proportion of Treg cell was elevated (P<0.05). The trend of corresponding indexes in the LPS group was opposite to the above (P<0.05). Compared with the high-dose Zhike Pingchuan Formula group, the pathological damage of lung tissue increased in the high-dose Zhike Pingchuan Formula+LPS group. The proliferation of bronchial epithelial goblet cells and the secretion of mucus increased, the IgE concentration, TNF-α, IL-17 levels, Th17 cell proportion, Th17/Treg ratio and TLR4, TRAF6, p-NF-κB p65 protein expression increased, while the Treg cell proportion decreased (P<0.05). Conclusion Zhike Pingchuan Formula may reduce airway inflammation in BA mice by inhibiting TLR4/TRAF6/NF-κB pathway.

Key words: asthma, Zhike Pingchuan Formula, toll-like receptor 4/tumor necrosis factor receptor-associated factor 6/nuclear factor kappa-B pathway, airway inflammation

CLC Number:

R256.12

Figures/Tables 7

References 19

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组别	TNF-α	IL-17
对照组	8.66±0.31	15.52±0.41
BA组	34.95±1.23^a	58.61±2.06^a
止咳平喘方低剂量组	29.51±1.01^b	47.87±1.85^b
止咳平喘方高剂量组	12.88±0.42^bc	20.66±1.31^bc
地塞米松组	12.91±0.43^bc	20.72±1.35^bc
LPS组	43.35±1.87^b	67.68±2.73^b
止咳平喘方高剂量+LPS组	25.54±0.96^d	43.36±2.05^d
F	1 906.643^＊＊	1 535.875^＊＊

组别	TNF-α	IL-17
对照组	8.66±0.31	15.52±0.41
BA组	34.95±1.23^a	58.61±2.06^a
止咳平喘方低剂量组	29.51±1.01^b	47.87±1.85^b
止咳平喘方高剂量组	12.88±0.42^bc	20.66±1.31^bc
地塞米松组	12.91±0.43^bc	20.72±1.35^bc
LPS组	43.35±1.87^b	67.68±2.73^b
止咳平喘方高剂量+LPS组	25.54±0.96^d	43.36±2.05^d
F	1 906.643^＊＊	1 535.875^＊＊

组别	Th17/%	Treg/%	Th17/Treg比值
对照组	3.19±0.12	33.56±1.64	0.09±0.01
BA组	22.67±1.01^a	12.28±0.56^a	1.85±0.06^a
止咳平喘方低剂量组	16.78±0.73^b	17.73±0.61^b	0.95±0.05^b
止咳平喘方高剂量组	7.54±0.31^bc	29.01±1.15^bc	0.26±0.02^bc
地塞米松组	7.52±0.19^bc	29.22±1.18^bc	0.25±0.02^bc
LPS组	26.67±1.21^b	9.66±0.31^b	2.76±0.13^b
止咳平喘方高剂量+LPS组	14.46±0.68^d	20.23±1.02^d	0.71±0.06^d
F	857.346^＊＊	486.504^＊＊	1 487.178^＊＊

组别	Th17/%	Treg/%	Th17/Treg比值
对照组	3.19±0.12	33.56±1.64	0.09±0.01
BA组	22.67±1.01^a	12.28±0.56^a	1.85±0.06^a
止咳平喘方低剂量组	16.78±0.73^b	17.73±0.61^b	0.95±0.05^b
止咳平喘方高剂量组	7.54±0.31^bc	29.01±1.15^bc	0.26±0.02^bc
地塞米松组	7.52±0.19^bc	29.22±1.18^bc	0.25±0.02^bc
LPS组	26.67±1.21^b	9.66±0.31^b	2.76±0.13^b
止咳平喘方高剂量+LPS组	14.46±0.68^d	20.23±1.02^d	0.71±0.06^d
F	857.346^＊＊	486.504^＊＊	1 487.178^＊＊

组别	TLR4/ GAPDH	TRAF6/ GAPDH	p-NF-κB p65/ NF-κB p65
对照组	0.65±0.04	0.41±0.03	0.23±0.01
BA组	1.84±0.22^a	0.86±0.03^a	0.64±0.05^a
止咳平喘方低剂量组	1.50±0.18^b	0.71±0.05^b	0.50±0.04^b
止咳平喘方高剂量组	1.03±0.11^bc	0.52±0.03^bc	0.31±0.03^bc
地塞米松组	1.01±0.12^bc	0.51±0.04^bc	0.32±0.02^bc
LPS组	2.29±0.23^b	0.94±0.04^b	0.81±0.07^b
止咳平喘方高剂量+LPS组	1.38±0.12^d	0.69±0.05^d	0.47±0.03^d
F	73.344^＊＊	145.358^＊＊	155.540^＊＊