Pingchuan formula improve airway remodeling via regulation of the nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3/interleukin-1β and C-Jun N-terminal kinase pathways.

Abstract

Asthma is a prevalent chronic inflammatory airway disease that affects both adults and children. Inflammation-induced airway remodeling can lead to irreversible damage to the airways. Traditional Chinese medicine (TCM) plays a significant role in healthcare, offering potential improvements for chronic airway inflammation associated with asthma. The objective of this study is to investigate the efficacy of Pingchuan formula (PCF) in treating asthma and explore its underlying mechanisms. The asthmatic model mice were sensitized on days 1 and 7, followed by aerosolized OVA challenge for a total of 21 times starting from day 14, spanning weeks 3 to 9. PCF was administered daily after the first challenge. The mice were orally dosed daily based on their individual body weights for a continuous period of 47 days. The examined items encompassed proteomic analysis of the target proteins impacted by PCF. The levels of interleukins IL-1β, IL-18, and transforming growth factor-β (TGF-β) in bronchoalveolar lavage fluid (BALF) were measured using ELISA. Immunohistochemistry was employed to assess the expression levels of nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3), caspase-1, and TGF-β in the airways. Western blotting and real-time quantitative PCR analysis were conducted to determine NLRP3 and caspase-1 expression levels. Additionally, Western blotting was performed to evaluate C-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), matrix metalloproteinase-9 (MMP-9), N-cadherin, E-cadherin, and tenascin C (T-NC) expression. Compared to the asthma model group, histological analysis using hematoxylin and eosin (HE) staining and Masson staining revealed that PCF exhibited a significant reduction in airway inflammation exudation and collagen fiber production in asthmatic mice. The proteomics analysis revealed that there were 174 proteins exhibiting differential expression in the PCF group compared to the asthma model group, indicating a strong association with negative regulation of the cell cycle and inflammation in the airways of asthmatic individuals. The PCF also exhibited a significant reduction in the protein and mRNA expressions of NLRP3 and caspase-1 in lung tissue (P<0.05). Additionally, it decreased the protein expressions of ASC, p-JNK, MMP-9, E-cadherin, and T-NC while increasing N-cadherin levels (P<0.05). The inflammatory factors IL-1β, IL-18 and TGF-β were reduced (P<0.05). This study revealed that Pingchuan Decoction can inhibit airway remodeling by inhibiting NLRP/IL-1β and JNK pathway activation, and effectively improve airway histological inflammation and remodeling in mice.