Deleterious effects of intestinal ischemia/reperfusion injury in the mouse enteric nervous system are associated with protein nitrosylation

Abstract

Changes in intestinal function, notably impaired transit, following ischemia/reperfusion (I/R) injury are likely to derive, at least in part, from damage to the enteric nervous system. Currently, there is a lack of quantitative data and methods on which to base quantitation of changes that occur in enteric neurons. In the present work, we have investigated quantifiable changes in response to ischemia of the mouse small intestine followed by reperfusion from 1 h to 7 days. I/R caused distortion of nitric oxide synthase (NOS)-containing neurons, the appearance of a TUNEL reaction in neurons, protein nitrosylation and translocation of Hu protein. Protein nitrosylation was detected after 1 h and was detectable in 10% of neurons by 6 h in the ischemic region, indicating that reactive peroxynitrites are rapidly produced and can interact with proteins soon after reperfusion. Apoptosis, revealed by TUNEL staining, was apparent at 6 h. The profile sizes of NOS neurons were increased by 60% at 2 days and neurons were still swollen at 7 days, both in the ischemic region and proximal to the ischemia. The distribution of the enteric neuron marker and oligonucleotide binding protein, Hu, was significantly changed in both regions. Hu protein translocation to the nucleus was apparent by 3 h and persisted for up to 7 days. Particulate Hu immunoreactivity was observed in the ganglia 3 h after I/R but was never observed in control. Our observations indicate that effects of I/R injury can be detected after 1 h and that neuronal changes persist to at least 7 days. Involvement of NO and reactive oxygen species in the changes is indicated by the accumulation of nitrosylated protein aggregates and the swelling and distortion of nitrergic neurons. It is concluded that damage to the enteric nervous system, which is likely to contribute to functional deficits following ischemia and re-oxygenation in the intestine, can be quantified by Hu protein translocation, protein nitrosylation, swelling of nitrergic neurons and apoptosis.

Influence of β-galacto-oligosaccharide on growth performance and components of intestinal barrier in broilers during heat stress

“…Intestinal mucosal lesions are among the initial manifestations of stress, and are attributed to ischemia of the gut wall, production of reactive oxygen species (Lambert et al, 2009), expression of heat shock proteins, and increases in pro-inflammatory cytokines and intestinal microbial dysbiosis (Song et al, 2014). Shedding of epithelial cells in response to these changes might have accounted for the decreased villus surface area and lower VH : CD observed in the current study (Rivera et al, 2011). An increase in villus surface area was observed in the duodenum with 0.1%β-GOS, in the jejunum with 0.2 and 0.5%β-GOS, and in the ileum in all the β-GOS-supplemented heatstressed groups.…”

Heat shock protein 70 mRNA expression and immune response of heat-stressed finishing broilers fed propolis (bee glue) supplementation

“…Several possible reasons exist as to why intestinal morphology was changed in response to HS, including lesions induced by intestinal ischemia and the presence of toxins (Xu et al, 2009). Ischemia of the enteric canal can cause epithelial shedding (Rivera et al, 2011), proceeding to shortened villus height and deeper crypt depth. Jia et al (2010) found that both villus height and crypt depth are important indicators of the digestive health of poultry and are directly related to the absorptive capacity of the mucous membrane.…”

Resveratrol Reduces Morphologic Changes in the Myenteric Plexus and Oxidative Stress in the Ileum in Rats with Ischemia/Reperfusion Injury

“…The reperfusion period in the present study was 3 h, whereas previous studies reported that neuronal death by apoptosis, the leading cause The results are expressed as neurons per cm 2 (HuC/D and nNOS) or glial cells per cm 2 (S100). The values represent the mean ± standard error, five animals per group Dig Dis Sci of neuronal death following I/R, begins after 6 h of reperfusion [8,44].…”