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The Effect of Ischemia and Reperfusion on Enteric Glial Cells and Contractile Activity in the Ileum

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Abstract

Background

We investigated the effects of ischemia followed by different periods of reperfusion (I/R) on immunoreactive S100β-positive glial and Hu-immunoreactive neurons co-expressing the P2X2 receptor in the myenteric plexus of the rat ileum.

Methods

The ileal artery was occluded for 35 min with an atraumatic vascular clamp. The animals were killed 24 h, 72 h, and 1 week after ischemia. Sham animals were not submitted to ileal artery occlusion. The relative density, size, and co-localization of P2X2 receptor-expressing cells in relation to S100β-immunoreactive glial and Hu-immunoreactive neuronal cells were evaluated. Additionally, we analyzed the effects of I/R on gastrointestinal transit and ileum contractile activity.

Results

The cellular density of P2X2 receptor and neuronal Hu immunoreactivity/cm2 decreased after I/R, whereas glial S100β immunoreactivity/cm2 increased. No significant differences between sham and I/R groups were observed regarding the perikarya area of Hu-positive neurons. The area of S100β-immunoreactive glial cells increased by 24.1 % 1 week after I/R compared with the 24 h group. Methylene blue progression along the small intestine decreased (P < 0.05) from 24.5 ± 2.3 % in the sham group to 17.2 ± 2.0 % 1 week post-ischemia. We noted a significant (P < 0.05) decrease in the maximal contraction amplitude triggered by electrical field stimulation in the presence of ATP in preparations submitted to 24 h of I/R.

Conclusions

Changes in the P2X2 receptor density parallel myenteric neuronal loss following I/R of the rat ileum. This, together with the increase in the activated (oversized) glial cells, may contribute to decreased GI motility after I/R.

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Acknowledgments

We would like to thank Professors Jackson Cioni Bittencourt and Carol Fuzeti Elias for the use of their fluorescence microscope and Rosana Prisco for statistical analysis. These studies were supported by (FAPESP/São Paulo Foundation Research/2008/05314-5, 2008/08728-5, 2012/06434-0; 2012/00259-1). The experiments performed at ICBAS-UP were partially supported by FCT (FCT project Pest-OE/SAU/UI215/2014) with the participation of FEDER funding. Cátia Vieira and Isabel Silva were in receipt of Ph.D. studentships from FCT (SFRH/BD/79091/2011 and SFRH/BD/88855/2012, respectively).

Conflict of interest

The authors have no conflict of interest.

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Authors and Affiliations

  1. Department of Anatomy, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Dr. Lineu Prestes, 2415, CEP 05508-900, São Paulo, Brazil

    Cristina Eusébio Mendes, Kelly Palombit & Patricia Castelucci

  2. Laboratory of Pharmacology and Neurobiology, Center for Drug Discovery and Innovative Medicines (MedInUP), Instituto de Ciências Biomédicas de Abel Salazar, University of Porto (ICBAS-UP), Porto, Portugal

    Cátia Vieira, Isabel Silva & Paulo Correia-de-Sá

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  1. Cristina Eusébio Mendes
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  6. Patricia Castelucci
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Correspondence to Patricia Castelucci.

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Mendes, C.E., Palombit, K., Vieira, C. et al. The Effect of Ischemia and Reperfusion on Enteric Glial Cells and Contractile Activity in the Ileum. Dig Dis Sci 60, 2677–2689 (2015). https://doi.org/10.1007/s10620-015-3663-3

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