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Identification of dihalogenated proteins in rat intestinal mucosa injured by indomethacin.

Takagi T, Naito Y, Okada H, Okayama T, Mizushima K, Yamada S, Fukumoto K, Inoue K, Takaoka M, Oya-Ito T, Uchiyama K, Ishikawa T, Handa O, Kokura S, Yagi N, Ichikawa H, Kato Y, Osawa T, Yoshikawa T - J Clin Biochem Nutr (2011)

Bottom Line: Tissue-associated MPO activity was measured in the intestinal mucosa as an index of neutrophil infiltration.Single administration of indomethacin elicited increased ulcerative area and MPO activity in the small intestine. 2D-PAGE showed an increased level of DiBrY-modified proteins in the indomethacin-induced injured intestinal mucosa and 6 modified proteins were found.Enolase-1 and albumin were found to be DiBrY modified.

View Article: PubMed Central - PubMed

Affiliation: Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.

ABSTRACT
Previous studies have shown that activated neutrophils and their myeloperoxidase (MPO)-derived products play a crucial role in the pathogenesis of non-steroidal anti-inflammatory drug (NSAID)-related small intestinal injury. The aim of the present study is to identify dihalogenated proteins in the small intestine on indomethacin administration. Intestinal damage was induced by subcutaneous administration of indomethacin (10 mg/kg) in male Wistar rats, and the severity of the injury was evaluated by measuring the area of visible ulcerative lesions. Tissue-associated MPO activity was measured in the intestinal mucosa as an index of neutrophil infiltration. The dihalogenated proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) using novel monoclonal antibodies against dibromotyrosine (DiBrY), and they were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) peptide mass fingerprinting and a Mascot database search. Single administration of indomethacin elicited increased ulcerative area and MPO activity in the small intestine. 2D-PAGE showed an increased level of DiBrY-modified proteins in the indomethacin-induced injured intestinal mucosa and 6 modified proteins were found. Enolase-1 and albumin were found to be DiBrY modified. These proteins may be responsible for the development of neutrophil-associated intestinal injury induced by indomethacin.

No MeSH data available.


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Representative macroscopic findings and ulcer index after indomethacin administration. To determine the extent of intestinal injury, 1% Evans blue was injected intravenously 30 min before euthanasia. The representative macroscopic findings indicate that the small intestinal injuries were induced 24 h after indomethacin administration (B); a normal untreated intestine is shown in (A). The ulcer index after indomethacin administration was evaluated as described in Materials and Methods (C). Data represent the mean ± (SEM) of 7 rats. *p<0.01 compared to the sham group (0 h).
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Figure 1: Representative macroscopic findings and ulcer index after indomethacin administration. To determine the extent of intestinal injury, 1% Evans blue was injected intravenously 30 min before euthanasia. The representative macroscopic findings indicate that the small intestinal injuries were induced 24 h after indomethacin administration (B); a normal untreated intestine is shown in (A). The ulcer index after indomethacin administration was evaluated as described in Materials and Methods (C). Data represent the mean ± (SEM) of 7 rats. *p<0.01 compared to the sham group (0 h).

Mentions: A single administration of 10 mg/kg indomethacin elicited multiple erosions in the small intestine (Fig. 1B); a normal untreated small intestine is shown in Fig. 1A for comparison. The ulcer index gradually increased with time, and significant increases in this index were noted 6, 12, and 24 h after indomethacin administration (Fig. 1C).


Identification of dihalogenated proteins in rat intestinal mucosa injured by indomethacin.

Takagi T, Naito Y, Okada H, Okayama T, Mizushima K, Yamada S, Fukumoto K, Inoue K, Takaoka M, Oya-Ito T, Uchiyama K, Ishikawa T, Handa O, Kokura S, Yagi N, Ichikawa H, Kato Y, Osawa T, Yoshikawa T - J Clin Biochem Nutr (2011)

Representative macroscopic findings and ulcer index after indomethacin administration. To determine the extent of intestinal injury, 1% Evans blue was injected intravenously 30 min before euthanasia. The representative macroscopic findings indicate that the small intestinal injuries were induced 24 h after indomethacin administration (B); a normal untreated intestine is shown in (A). The ulcer index after indomethacin administration was evaluated as described in Materials and Methods (C). Data represent the mean ± (SEM) of 7 rats. *p<0.01 compared to the sham group (0 h).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3045693&req=5

Figure 1: Representative macroscopic findings and ulcer index after indomethacin administration. To determine the extent of intestinal injury, 1% Evans blue was injected intravenously 30 min before euthanasia. The representative macroscopic findings indicate that the small intestinal injuries were induced 24 h after indomethacin administration (B); a normal untreated intestine is shown in (A). The ulcer index after indomethacin administration was evaluated as described in Materials and Methods (C). Data represent the mean ± (SEM) of 7 rats. *p<0.01 compared to the sham group (0 h).
Mentions: A single administration of 10 mg/kg indomethacin elicited multiple erosions in the small intestine (Fig. 1B); a normal untreated small intestine is shown in Fig. 1A for comparison. The ulcer index gradually increased with time, and significant increases in this index were noted 6, 12, and 24 h after indomethacin administration (Fig. 1C).

Bottom Line: Tissue-associated MPO activity was measured in the intestinal mucosa as an index of neutrophil infiltration.Single administration of indomethacin elicited increased ulcerative area and MPO activity in the small intestine. 2D-PAGE showed an increased level of DiBrY-modified proteins in the indomethacin-induced injured intestinal mucosa and 6 modified proteins were found.Enolase-1 and albumin were found to be DiBrY modified.

View Article: PubMed Central - PubMed

Affiliation: Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.

ABSTRACT
Previous studies have shown that activated neutrophils and their myeloperoxidase (MPO)-derived products play a crucial role in the pathogenesis of non-steroidal anti-inflammatory drug (NSAID)-related small intestinal injury. The aim of the present study is to identify dihalogenated proteins in the small intestine on indomethacin administration. Intestinal damage was induced by subcutaneous administration of indomethacin (10 mg/kg) in male Wistar rats, and the severity of the injury was evaluated by measuring the area of visible ulcerative lesions. Tissue-associated MPO activity was measured in the intestinal mucosa as an index of neutrophil infiltration. The dihalogenated proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) using novel monoclonal antibodies against dibromotyrosine (DiBrY), and they were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) peptide mass fingerprinting and a Mascot database search. Single administration of indomethacin elicited increased ulcerative area and MPO activity in the small intestine. 2D-PAGE showed an increased level of DiBrY-modified proteins in the indomethacin-induced injured intestinal mucosa and 6 modified proteins were found. Enolase-1 and albumin were found to be DiBrY modified. These proteins may be responsible for the development of neutrophil-associated intestinal injury induced by indomethacin.

No MeSH data available.


Related in: MedlinePlus