Jinling Luan, Xiaoyan Su, Na Chen
RAM may attenuate allergic asthma by inhibiting the TNF-mediated necroptosis pathway. This study provides a pharmacological basis for RAM as a promising therapeutic agent for asthma treatment.
OBJECTIVE: The aim of thev study is to investigate the therapeutic effects and underlying mechanisms of RAM in a house dust mite (HDM)-induced murine model of allergic asthma, with a focus on necroptosis and TNF signaling.
MATERIAL AND METHODS: Allergic asthma was induced in mice and HBE cells by HDM challenge. Animals were treated with RAM to assess its effects on airway inflammation, hyperresponsiveness, IgE production, and cytokine/chemokine expression. In vitro, necroptosis markers (RIPK1, RIPK3, p-MLKL), inflammatory cytokines, and cell viability were evaluated in HBE cells. Network pharmacology and molecular docking were employed to predict RAM's bioactive compounds and their primary targets, with a focus on the necroptosis pathway. The role of TNF was further validated through overexpression experiments in HBE cells.
RESULTS: RAM treatment significantly alleviated asthma phenotypes, reducing inflammatory cell infiltration in BALF, serum IgE levels, airway hyperresponsiveness, and pulmonary expression of Cxcl1/Cxcl2. RAM suppressed expression of Ripk1, Ripk3, p-MLKL, and caspase-3, alongside reduced proinflammatory cytokines (IL-1α, IL-1β, IL-33), in HDM-induced mice and HBE cells. Network pharmacology identified TNF as a top-ranked target within the necroptosis pathway, and molecular docking confirmed binding affinities between TNF and five RAM compounds. Experimentally, RAM downregulated TNF expression in mouse lungs and HBE cells. TNF overexpression reversed RAM's protective effects, restoring Ripk1/Ripk3 expression and diminishing cell viability.
CONCLUSIONS: RAM may attenuate allergic asthma by inhibiting the TNF-mediated necroptosis pathway. This study provides a pharmacological basis for RAM as a promising therapeutic agent for asthma treatment.