Molecular effects of intermittent stress on primary feline uroepithelial cell culture as an in vitro model of feline idiopathic cystitis

Patrícia Hatala; Csilla Sebők; Máté Mackei; Karina Kárpáti; Péter Gálfi; Zsuzsanna Neogrády; Gábor Mátis

doi:10.3389/fvets.2023.1258375

Molecular effects of intermittent stress on primary feline uroepithelial cell culture as an in vitro model of feline idiopathic cystitis

Front Vet Sci. 2023 Nov 6:10:1258375. doi: 10.3389/fvets.2023.1258375. eCollection 2023.

Authors

Patrícia Hatala¹, Csilla Sebők¹, Máté Mackei^{1

2}, Karina Kárpáti¹, Péter Gálfi³, Zsuzsanna Neogrády¹, Gábor Mátis^{1

2}

Affiliations

¹ Division of Biochemistry, Department of Physiology and Biochemistry, University of Veterinary Medicine Budapest, Budapest, Hungary.
² National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, University of Veterinary Medicine Budapest, Budapest, Hungary.
³ Department of Pharmacology and Toxicology, University of Veterinary Medicine Budapest, Budapest, Hungary.

Abstract

Introduction: The most common cause of feline lower urinary tract disease (FLUDT) is feline idiopathic cystitis (FIC), which is a complex multifactorial disease with symptoms including stranguria, dysuria, hematuria, and pain during urination. The development of these symptoms is often triggered by stress, and in case of chronic stress, these symptoms will many times return. One of the most important stress hormones in the pathogenesis of FIC is norepinephrine (NE), as persistently elevated level of this hormone can be measured in the blood of cats with FIC. However, it is not well understood if recurrently elevated level of NE has any direct effect on urinary bladder, therefore the aim of this study was to investigate the molecular effects of intermittent NE exposure on feline primary uroepithelial cell culture.

Methods: Primary uroepithelial cells were gained from the mucosa of the bladder of a euthanized cat, and were cultured for 6 days, then they were exposed to 10, 100, and 1,000 μM NE treatment for 3 × 1 h, including a 1 h long regeneration period between exposures.

Results: NE was able to trigger pro-inflammatory response and oxidative stress in the uroepithelial cells by increasing the level of stromal cell derived factor 1 (SDF-1) and H₂O₂ in cell culture media. In addition, NE increased the permeability of the uroepithelium, since decreased glycosaminoglycan (GAG) concentration, tight junction protein claudin-4 content, and TER values were measured after the NE treatments.

Discussion: Based on these results it can be concluded that recurrent stress mimicked by 3×1 h NE treatment has a direct molecular effect on the uroepithelial cells, which leads to inflammatory response, oxidative stress and decreased barrier function of the uroepithelium. Therefore, intermittent release of NE may have an important role in the pathogenesis of FIC and the results of this study may contribute to a better understanding of the development of this illness.

Keywords: FIC; chronic stress; norepinephrine; uroepithelial barrier; uroepithelial cells.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Project no. RRF-2.3.1-21-2022-00001 has been implemented with the support provided by the Recovery and Resilience Facility (RRF), financed under the National Recovery Fund budget estimate, RRF-2.3.1-21 funding scheme.