• Home
  • Search Results
  • Effects of nerve growth factor antagonist K252a on peritoneal mast cell degranulation: implications for rat postoperative ileus.

Effects of nerve growth factor antagonist K252a on peritoneal mast cell degranulation: implications for rat postoperative ileus.

American journal of physiology. Gastrointestinal and liver physiology (2015-09-26)
Sergio Berdún, Jakub Rychter, Patri Vergara
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
Sodium bicarbonate, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Sodium bicarbonate, ACS reagent, ≥99.7%
Sigma-Aldrich
Hexadecyltrimethylammonium bromide, ≥98%
Sigma-Aldrich
Sodium carbonate, ACS reagent, anhydrous, ≥99.5%, powder or granules
Sigma-Aldrich
Hexadecyltrimethylammonium bromide, for molecular biology, ≥99%
Sigma-Aldrich
Sodium carbonate, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium carbonate, powder, ≥99.5%, ACS reagent
Sigma-Aldrich
Histopaque®-1119, sterile-filtered, density: 1.119 g/mL
Sigma-Aldrich
Sodium bicarbonate, ReagentPlus®, ≥99.5%, powder
Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Sodium carbonate, ReagentPlus®, ≥99.5%
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥99%
Sigma-Aldrich
Sodium bicarbonate, BioXtra, 99.5-100.5%
Sigma-Aldrich
Sodium carbonate, ACS reagent (primary standard), anhydrous, 99.95-100.05% dry basis
Sigma-Aldrich
Hexadecyltrimethylammonium bromide, BioXtra, ≥99%
Sigma-Aldrich
Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Compound 48/80
Sigma-Aldrich
Sodium bicarbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.7%
Sigma-Aldrich
Hexadecyltrimethylammonium bromide, BioUltra, for molecular biology, ≥99.0% (AT)
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥98% (TLC)
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥98.0% (NT)
Sigma-Aldrich
Myeloperoxidase from human leukocytes, lyophilized powder, ≥50 units/mg protein
Sigma-Aldrich
NGF-β from rat, recombinant, expressed in Sf21 cells, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Sodium carbonate, puriss., meets analytical specification of Ph. Eur., BP, NF, FCC, E500, anhydrous, 99.5-100.5% (calc. to the dried substance)
Sigma-Aldrich
Sodium bicarbonate, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E500, 99.0-100.5%, powder
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, tablet, 1 mg substrate per tablet
Sigma-Aldrich
Sodium bicarbonate, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.7%
Sigma-Aldrich
Sodium hydrogencarbonate, −40-+140 mesh, ≥95%