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Updated: Apr 3, 2026
Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays
Published on: July 4, 2018
Effects of nerve growth factor antagonist K252a on peritoneal mast cell degranulation: implications for rat
Sergio Berdún1, Jakub Rychter2, Patri Vergara3
1Department of Cell Biology, Physiology and Immunology, Veterinary School, Autonomous University of Barcelona, Barcelona, Spain; Neuroscience Institute, Autonomous University of Barcelona, Barcelona, Spain; and.
Abstract:
Stabilization of mast cell (MC) degranulation has been proposed to prevent postoperative ileus (POI). Nerve growth factor (NGF) mediates MC degranulation. The aim of the study was to evaluate whether NGF receptor antagonist K252a acts as a MC stabilizer in vitro and in vivo model of POI. Peritoneal mast cells (PMCs) were obtained from Sprague-Dawley rats and were incubated with K252a and exposed to NGF or Compound 48/80 (C48/80). MC degranulation was assessed by β-hexosaminidase assay. POI was induced in rats by intestinal manipulation (IM). Rats were pretreated with K252a (100 μg/kg sc) 20 min prior to POI induction. At 20 min after IM, release of rat mast cell protease 6 (RMCP-6) was evaluated in peritoneal lavage. At 24 h, intestinal transit (IT) and gastric emptying (GE) were evaluated. Ileal inflammation was assessed by myeloperoxidase (MPO) activity, expression of IL-6, NGF, TrkA, RMCP-2 and 6, and MC density within the full-thickness ileum. C48/80 and NGF evoked degranulation of PMCs in a dose-dependent manner. K252a prevented NGF-evoked, but not C48/80-evoked, MC degranulation. IM evoked the release of peritoneal RMCP-6 and subsequently delayed IT and GE. IM increased MPO activity and expression of IL-6. In IM rats, K252a prevented upregulation of IL-6 expression and reduced TrkA. IT, GE, and inflammation were not affected by K252a. K252a inhibited NGF-evoked degranulation of PMCs in vitro. In vivo, K252a decreased IL-6 and PMC degranulation. This may be of relevance for the development of new therapeutic targets for POI.
Insights
Nerve growth factor (NGF) antagonist K252a stabilizes mast cells (MCs) by preventing NGF-induced degranulation. While K252a reduced IL-6 and MC degranulation in a postoperative ileus model, it did not improve intestinal transit or gastric emptying.
Area of Science:
- Gastroenterology
- Immunology
- Pharmacology
Background:
- Postoperative ileus (POI) is a common complication, and mast cell (MC) degranulation is implicated in its pathogenesis.
- Nerve growth factor (NGF) plays a role in mediating MC degranulation.
Purpose of the Study:
- To investigate the efficacy of K252a, an NGF receptor antagonist, as a mast cell stabilizer in an in vitro and in vivo model of POI.
Main Methods:
- Peritoneal mast cells (PMCs) were isolated from rats and treated with K252a, NGF, or Compound 48/80 (C48/80). MC degranulation was measured via beta-hexosaminidase assay.
- POI was induced in rats by intestinal manipulation (IM). Rats were pretreated with K252a before IM. Parameters assessed included peritoneal RMCP-6 release, intestinal transit (IT), gastric emptying (GE), ileal inflammation (MPO activity, IL-6), and MC markers.
Main Results:
- K252a inhibited NGF-induced, but not C48/80-induced, PMC degranulation in vitro.
- In vivo, K252a treatment reduced IL-6 expression and peritoneal MC degranulation following IM.
- K252a did not significantly improve IT, GE, or reduce overall ileal inflammation (MPO activity).
Conclusions:
- K252a demonstrates mast cell stabilizing properties by inhibiting NGF-mediated degranulation.
- The observed reduction in IL-6 and MC degranulation suggests a potential role for K252a in modulating inflammatory pathways in POI.
- Further research is needed to explore the therapeutic potential of K252a for POI, as current findings did not translate to functional recovery of IT and GE.
