Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=41341025
&cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests
in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215
Glial cell line-derived neurotrophic factor inhibits mast-cell-like RBL-2H3 cells
activation via Ca2+-mediated degranulation and Ca2+/CaMK?/JNK pathway
#MMPMID41341025
Huang W
; Zeng L
; Zhang L
; Zhang X
; Xie Q
Front Pharmacol
2025[]; 16
(?): 1697815
PMID41341025
show ga
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 ?-hexosaminidase release
assays. Culture supernatants were analyzed for TNF?, IL-1?, and IL-6 secretion
using ELISA. Key signaling molecules-GDNF family receptor ?1 (GFR?1), receptor
Tyrosine Kinase (RET), calcium/calmodulin-dependent protein kinase II (CaMK?),
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-GFR?1/RET complexes and CaMK?-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 activated RBL-2H3 cells after GDNF treatment.
Immunofluorescence results demonstrated co-localization of GDNF with GFR?1 on
RBL-2H3 cell membranes and CaMKII with JNK in the cytoplasm. There were
interactions between GDNF and GFR?1/RET, as well as CaMK II and JNK. RET
inhibitor eliminated this inhibitory effect of GDNF on RBL-2H3 cell degranulation
and inflammatory factor release. Ca2+ chelator and CaMK? RNAi had the same
inhibitory effect on degranulation, release of inflammatory cytokines and
phosphorylation of JNK. However, in their presence, GDNF had no additional
inhibitory effect. CONCLUSION: GDNF can decrease the intracellular concentration
of Ca(2+) in activated RBL-2H3 cells by attaching to GFL?1/RET receptors located
on the membrane of RBL-2H3 cells, subsequently inhibiting Ca(2+)-mediated
degranulation and the Ca(2+)/CaMKII/JNK pathway responsible for the release of
inflammatory cytokines.
free
free
free
