Glial cell line-derived neurotrophic factor inhibits mast-cell-like RBL-2H3 cells activation via Ca2+-mediated degranulation and Ca2+/CaMKⅡ/JNK pathway

作者：Wei Huang, Li Zeng, Li Zhang, Xinxing Zhang, Qin Xie · 发表于：Frontiers in Pharmacology · 年份：2025 · DOI：10.3389/fphar.2025.1697815 · 被引用次数：1 · 研究领域：Nerve injury and regeneration、Mast cells and histamine、Protein Kinase Regulation and GTPase Signaling

Introduction Mast cells are important component of the intestinal immune system, play a crucial role in the pathogenesis of inflammatory bowel disease. Glial cell line-derived neurotrophic factor (GDNF), as a multifunctional growth factor, has recently garnered attention for its role in the inhibition of mast cells activation. This study aims to explore the potential mechanisms by which GDNF inhibits mast cell activation. Methods In this study, RBL-2H3 cells were used as an in vitro cell model of mast cells, which were cultured and treated with various interventions prior to collection of cells and culture supernatants. IgE-mediated degranulation were evaluated through β-hexosaminidase release assays. Culture supernatants were analyzed for TNFα, IL-1β, and IL-6 secretion using ELISA. Key signaling molecules—GDNF family receptor α1 (GFRα1), receptor Tyrosine Kinase (RET), calcium/calmodulin-dependent protein kinase II (CaMKⅡ), total and phosphorylated c-Jun N-terminal kinase (JNK), and JNK isoforms—were quantified at mRNA and protein levels using Quantitative Real-time polymerase chain reaction and Western blot. Intracellular Ca 2+ were monitored fluorometrically. Immunofluorescence and protein binding assays were used to confirm interactions between GDNF-GFRα1/RET complexes and CaMKⅡ-JNK associations. Results GDNF inhibited the degranulation and release of inflammatory cytokines in activated RBL-2H3 cells. The intracellular Ca 2+ and the phosphorylation of JNK were reduced in...