[Exploration of therapeutic effect and mechanism of Jingfang Granules combined with dexamethasone on asthma based on metabolomics].

This study aims to investigate the ameliorative effects and underlying mechanisms of Jingfang Granules(JF) combined with dexamethasone(DEX) in asthmatic mice. Balb/c mice were randomly assigned to seven groups according to body weight: normal, model, low-dose JF(1 g·kg~(-1)·d~(-1)), medium-dose JF(2 g·kg~(-1)·d~(-1)), high-dose JF(4 g·kg~(-1)·d~(-1)), DEX(1 mg·kg~(-1)·d~(-1)), and a combination of JF(4 g·kg~(-1)·d~(-1)) and DEX(1 mg·kg~(-1)·d~(-1)). The asthma model was established by intraperitoneal injection of ovalbumin(OVA) and aluminum hydroxide. Serum immunoglobulin E(IgE) and lung interleukin-4(IL-4), interleukin-5(IL-5), and interleukin-13(IL-13) levels were determined by enzyme-linked immunosorbent assay(ELISA). White blood cells(WBC), eosinophils(EOS), and lymphocytes(LYM) in bronchoalveolar lavage fluid(BALF) were counted, and T helper cell 1(Th1)/T helper cell 2(Th2) ratios were analyzed by flow cytometry to assess the anti-inflammatory and immunomodulatory effects of JF combined with DEX in asthmatic mice. Non-targeted and targeted metabolomics analyses were conducted to explore therapeutic mechanisms. The activities of superoxide dismutase(SOD), catalase(CAT), and glutathione(GSH), as well as the level of malondialdehyde(MDA), were measured by ELISA, and mitochondrial morphology was observed by transmission electron microscopy. Western blot was used to detect the expression of ferroptosis-related proteins glutathione peroxidase 4(GPX4), solute carrier family 3 member 2(SLC3A2), solute carrier family 7 member 11(SLC7A11), and acyl-CoA synthetase long-chain family member 4(ACSL4), thereby validating the findings of the metabolomics study. The results showed that compared with the model group, JF combined with DEX significantly decreased serum IgE and lung IL-4, IL-5, and IL-13 levels, reduced WBC, EOS, and LYM counts in BALF, and restored Th1/Th2 balance. Additionally, the combination therapy increased SOD, CAT, and GSH levels, lowered the MDA level, improved mitochondrial ultrastructure, and normalized GPX4, SLC3A2, SLC7A11, and ACSL4 expression. It may also inhibit ferroptosis via the regulation of amino acid metabolism. In summary, JF combined with DEX exerts synergistic anti-inflammatory, immunomodulatory, and antioxidant effects. It inhibits ferroptosis in asthmatic mice probably by regulating amino acid metabolism pathways, thereby improving the pathological state of asthma. This study provides experimental evidence supporting the use of TCM compounds combined with glucocorticoids for asthma treatment and offers novel insights into their synergistic mechanisms of intervention.
Chronic respiratory disease
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Authors

Jiao Jiao, Yang Yang, Zhang Zhang, Pan Pan, Li Li, Dai Dai, Guo Guo, Lu Lu, Yao Yao
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