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AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms

INTRODUCTION AND OBJECTIVE: Excessive production of monocyte chemotactic protein-1 (MCP-1) has been observed in various inflammatory, chronic pain, and bladder overactive conditions. We have observed that over half of patients with interstitial cystitis/bladder pain syndrome (IC/BPS) express elevate...

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Autor principal: Luo, Yi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AME Publishing Company 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4708318/
http://dx.doi.org/10.3978/j.issn.2223-4683.2014.s045
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author Luo, Yi
author_facet Luo, Yi
author_sort Luo, Yi
collection PubMed
description INTRODUCTION AND OBJECTIVE: Excessive production of monocyte chemotactic protein-1 (MCP-1) has been observed in various inflammatory, chronic pain, and bladder overactive conditions. We have observed that over half of patients with interstitial cystitis/bladder pain syndrome (IC/BPS) express elevated MCP-1 in the urine. This study was to develop a mouse model that secretes MCP-1 by the urothelium to facilitate the study of IC/BPS in humans. METHODS: A 4.9 Kb transgene consisting of the uroplakin II gene promoter and mouse MCP-1 coding sequence with a secretory element was constructed and microinjected for developing a MCP-1 secreting transgenic mouse line (URO-MCP-1). Mice were screened by tail genotyping and backcrossed onto the C57BL/6 genetic background. RT-PCR was used to assess MCP-1 mRNAs in the bladder and ELISA used to assess MCP-1 protein in the urine. To induce bladder inflammation, lipopolysaccharide (LPS) was administered intravesically. Bladder inflammation was analyzed by histological hematoxylin and eosin (H&E). Von Frey filament stimulation was used to assess pelvic pain and micturition cages used to assess voiding dysfunction. Cadi-05, a novel Toll-like receptor modulator, was intradermally administered once daily for total three doses starting one day before cystitis induction up to day 1. The bladders were analyzed by histological H&E staining at day 2. RESULTS: URO-MCP-1 mice express MCP-1 mRNA in the bladder but not in other organs. Urine contains ~1,500 pg/mL of MCP-1. While the bladders of URO-MCP-1 mice were normal in the unmanipulated state, they exhibited a much lower threshold trigger for producing exaggerated responses to otherwise sub-noxious inflammatory stimuli such as LPS. Compared to control C57BL/6 mice, URO-MCP-1 mice exhibited clear histological bladder inflammation with intensive edema and cellular infiltration 24 hours after intravesical instillation of even a single low dose of LPS (1 µg in 100 µL). The bladder inflammation peaks at days 1-3 and lasts 5-7 days. URO-MCP-1 mice exhibited pelvic pain and increased urinary frequency and decreased urine output per void after cystitis induction (P<0.05). Intradermal Cadi-05 treatment significantly reduced bladder histopathology (P<0.01). CONCLUSIONS: The URO-MCP-1 model exhibits key symptomatology of IC/BPS and is responsive to intradermal Cadi-05 therapy. This model offers a unique translational opportunity for investigation of the pathological mechanisms and therapeutic development for the human disease.
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spelling pubmed-47083182016-01-26 AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms Luo, Yi Transl Androl Urol Podium Lecture INTRODUCTION AND OBJECTIVE: Excessive production of monocyte chemotactic protein-1 (MCP-1) has been observed in various inflammatory, chronic pain, and bladder overactive conditions. We have observed that over half of patients with interstitial cystitis/bladder pain syndrome (IC/BPS) express elevated MCP-1 in the urine. This study was to develop a mouse model that secretes MCP-1 by the urothelium to facilitate the study of IC/BPS in humans. METHODS: A 4.9 Kb transgene consisting of the uroplakin II gene promoter and mouse MCP-1 coding sequence with a secretory element was constructed and microinjected for developing a MCP-1 secreting transgenic mouse line (URO-MCP-1). Mice were screened by tail genotyping and backcrossed onto the C57BL/6 genetic background. RT-PCR was used to assess MCP-1 mRNAs in the bladder and ELISA used to assess MCP-1 protein in the urine. To induce bladder inflammation, lipopolysaccharide (LPS) was administered intravesically. Bladder inflammation was analyzed by histological hematoxylin and eosin (H&E). Von Frey filament stimulation was used to assess pelvic pain and micturition cages used to assess voiding dysfunction. Cadi-05, a novel Toll-like receptor modulator, was intradermally administered once daily for total three doses starting one day before cystitis induction up to day 1. The bladders were analyzed by histological H&E staining at day 2. RESULTS: URO-MCP-1 mice express MCP-1 mRNA in the bladder but not in other organs. Urine contains ~1,500 pg/mL of MCP-1. While the bladders of URO-MCP-1 mice were normal in the unmanipulated state, they exhibited a much lower threshold trigger for producing exaggerated responses to otherwise sub-noxious inflammatory stimuli such as LPS. Compared to control C57BL/6 mice, URO-MCP-1 mice exhibited clear histological bladder inflammation with intensive edema and cellular infiltration 24 hours after intravesical instillation of even a single low dose of LPS (1 µg in 100 µL). The bladder inflammation peaks at days 1-3 and lasts 5-7 days. URO-MCP-1 mice exhibited pelvic pain and increased urinary frequency and decreased urine output per void after cystitis induction (P<0.05). Intradermal Cadi-05 treatment significantly reduced bladder histopathology (P<0.01). CONCLUSIONS: The URO-MCP-1 model exhibits key symptomatology of IC/BPS and is responsive to intradermal Cadi-05 therapy. This model offers a unique translational opportunity for investigation of the pathological mechanisms and therapeutic development for the human disease. AME Publishing Company 2014-09 /pmc/articles/PMC4708318/ http://dx.doi.org/10.3978/j.issn.2223-4683.2014.s045 Text en 2014 Translational Andrology and Urology. All rights reserved.
spellingShingle Podium Lecture
Luo, Yi
AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms
title AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms
title_full AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms
title_fullStr AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms
title_full_unstemmed AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms
title_short AB45. URO-MCP-1 mice: a model demonstrating UCPPS hallmark symptoms
title_sort ab45. uro-mcp-1 mice: a model demonstrating ucpps hallmark symptoms
topic Podium Lecture
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4708318/
http://dx.doi.org/10.3978/j.issn.2223-4683.2014.s045
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