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Acute Oxidative Stress Affects IL-8 and TNF-α Expression in IPEC-J2 Porcine Epithelial Cells

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Abstract

Reactive oxygen species are implicated in cell and tissue damage in a number of diseases including acute and chronic inflammation of the gut. Effects of H2O2 exposure on non-carcinogenic porcine epithelial cell line, IPEC-J2 cells cultured on collagen-coated membrane inserts were monitored based on transepithelial electrical resistance (TER) change, extent of necrotic cell damage, gene expression of inflammatory cytokines IL-8 and TNF-α. Furthermore, the junction proteins claudin-1 and E-cadherin were also investigated by immunohistochemistry. Peroxide (1mM) increased IL-8 and TNF-α gene expression levels significantly allowing 1 h recovery time without affecting the cellular distribution of junction proteins, TER and cell survival rate. In conclusion, the IPEC-J2 cell line on membrane insert was introduced as a fast and reliable investigation tool for oxidative stimuli-triggered intestinal inflammation and in the future as a screening method for antioxidant and probiotic candidates.

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Acknowledgement

The research described here has been supported by the Hungarian Scientific Research Fund (grant OTKA no. 76133). We are indebted to Dr. Jody Gookin and Dr. Stephen Stauffer, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA, for providing IPEC-J2 cells and for the valuable advice on handling them. We also would to thank Dr. Adam Csordas, Division of Medical Biochemistry, Biocenter, Innbruck Medical University, Innsbruck, Austria, for critical reading of the manuscript. Besides useful practical and theoretical guidance, support in sequencing of PCR product (IL-8) from Dr. Balazs Gereben, Department of Endocrine Neurobiology, Institute of Experimental Medicine of the Hungarian Academy of Sciences, is also acknowledged.

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  1. Department of Pharmacology and Toxicology, Faculty of Veterinary Sciences, Szent István University, Budapest, István u. 2, 1078, Hungary

    Erzsebet Paszti-Gere, Edina Csibrik-Nemeth, Krisztina Szeker, Rita Csizinszky & Peter Galfi

  2. Department of Pathology and Forensic Veterinary Medicine, Faculty of Veterinary Sciences, Szent István University, Budapest, Hungary

    Csaba Jakab

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  1. Erzsebet Paszti-Gere
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Correspondence to Erzsebet Paszti-Gere.

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Paszti-Gere, E., Csibrik-Nemeth, E., Szeker, K. et al. Acute Oxidative Stress Affects IL-8 and TNF-α Expression in IPEC-J2 Porcine Epithelial Cells. Inflammation 35, 994–1004 (2012). https://doi.org/10.1007/s10753-011-9403-8

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