J Vet Sci.  2006 Jun;7(2):143-150. 10.4142/jvs.2006.7.2.143.

Alteration of nitrergic neuromuscular transmission as a result of acute experimental colitis in rat

• Sung TS
• La JH
• Kim TW
• Yang IS

• KMID: 1059200
• DOI: http://doi.org/10.4142/jvs.2006.7.2.143

Abstract

Nitric oxide (NO) is a non-adrenergic, non-cholinergic neurotransmitter found in the enteric nervous system that plays a role in a variety of enteropathies, including inflammatory bowel disease. Alteration of nitrergic neurons has been reported to be dependent on the manner by which inflammation is caused. However, this observed alteration has not been reported with acetic acid-induced colitis. Therefore, the purpose of the current study was to investigate changes in nitrergic neuromuscular transmission in experimental colitis in a rat model. Distal colitis was induced by intracolonic administration of 4% acetic acid in the rat. Animals were sacrificed at 4 h and 48 h postacetic acid treatment. Myeloperoxidase activity was significantly increased in the acetic acid-treated groups. However, the response to 60 mM KCl was not significantly different in the three groups studied. The amplitude of phasic contractions was increased by Nomega-nitro-L-arginine methyl ester (L-NAME) in the normal control group, but not in the acetic acid-treated groups. Spontaneous contractions disappeared during electrical field stimulation (EFS) in normal group. However, for the colitis groups, these contractions initially disappeared, and then reappeared during EFS. Moreover, the observed disappearance was diminished by L-NAME; this suggests that these responses were NO-mediated. In addition, the number of NADPH-diaphorase positive nerve cell bodies, in the myenteric plexus, was not altered in the distal colon; whereas the area of NADPH-diaphorase positive fibers, in the circular muscle layer, was decreased in the acetic acidtreated groups. These results suggest that NO-mediated inhibitory neural input, to the circular muscle, was decreased in the acetic acid-treated groups.

Keyword

colitis; electrical field stimulation; neuronal nitric oxide; nitrergic neuron; Nomega-nitro-L-arginine methyl ester

MeSH Terms

Acetic Acid/toxicity
Animals
Colitis/chemically induced/*pathology/*physiopathology
Colon/drug effects/enzymology/*innervation/pathology
Indicators and Reagents/toxicity
Male
Muscle Contraction/drug effects
Muscle, Smooth/drug effects/metabolism
Myenteric Plexus/pathology
NADPH Dehydrogenase/metabolism
NG-Nitroarginine Methyl Ester/pharmacology
Neuromuscular Junction/drug effects/*metabolism
Nitrergic Neurons/drug effects/*metabolism
Nitric Oxide/*metabolism
Peroxidase/metabolism
Potassium Chloride/pharmacology
Rats
Rats, Sprague-Dawley

Figure

• 

  Fig. 1 Colonic MPO activity in acetic acid-induced colitis rat model. The whole distal colon segment was measured in normal controls, at 4 and 48 h colitis group (n = 5). MPO activity was significantly increased in the colitis groups. Values are expressed as means ± SE. * p < 0.01.

• 

  Fig. 2 Tonic contraction of circular muscle in the distal colon was generated by 60 mM KCl. The maximum contractility of the circular muscle was not significantly altered by inflammation. Tension was expressed as mN per cross-sectional area (cm2) of tissue. Values are expressed as means ± SE (normal, n = 6; 4 h colitis, n = 7; 48 h colitis, n = 4).

• 

  Fig. 3 The effect of L-NAME on spontaneous contraction. Raw traces show that the amplitude of phasic contraction was increased after L-NAME administration in normal controls (n = 7, A), but not altered in either the 4 h (n = 5, B) or 48 h colitis groups (n = 6, C). The frequency of phasic contraction was unaffected by L-NAME in all groups studied (E). Values are expressed as means ± SE. * p < 0.05.

• 

  Fig. 4 The time to disappearance of spontaneous contraction during EFS. A. Raw traces show alteration of spontaneous contraction during EFS (8 Hz). The time to disappearance of spontaneous contraction (full lines) was slightly reduced in the 4 h colitis group (Ac) and significantly reduced in the 48 h colitis group (Ae) compared with that in the normal group (Aa). L-NAME prevented the disappearance of spontaneous contractions during EFS (Ab, d, f). B. These responses were not different in any of the groups studied (B, n = 5 for each group). Values are expressed as means ± SE. ▲: EFS on, ▼: EFS off.

• 

  Fig. 5 The number of NADPH-diaphorase positive nerve cell bodies in the myenteric plexus of the distal colon (A) normal controls, (B) 4 h colitis, (C) 48 h colitis, (n = 5 for each group). Scale bar = 20 µm.

• 

  Fig. 6 The number of NADPH-diaphorase positive nerve cell bodies per ganglion in the myenteric plexus. NADPH-diaphorase positive nerve cell bodies in the myenteric plexus was not affected by acetic acid treatment. Values are expressed as means ± SE.

• 

  Fig. 7 The area of NADPH-diaphorase positive nerve fibers in the circular muscle layer of the distal colon. (A) normal, (B) 4 h colitis, (C) 48 h colitis, (n = 5 for each group). The arrows indicate nerve fibers in the circular muscle layer. Scale bar = 100 µm.

• 

  Fig. 8 The area of NADPH-diaphorase positive nerve fibers in the circular muscle layer. The average of NADPH-diaphorase positive areas was slightly reduced in the 4 h colitis group (p = 0.06) and significantly reduced in the 48 h colitis group. Values are expressed as means ± SE. * p < 0.05.

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