ovalbumin and Cystitis--Interstitial

Topics: Animals; Antigens; Autoimmune Diseases; Cystitis; Cystitis, Interstitial; Cytokines; Disease Models, Animal; Gene Expression Regulation; Mice; Mice, Transgenic; Ovalbumin; Pelvic Pain; Urinary Bladder; Urination Disorders; Urothelium

Altered Toll-like receptor (TLR)4 activation has been identified in several chronic pain conditions but has not been well studied in interstitial cystitis/bladder pain syndrome (IC/BPS). Our previously published human studies indicated that patients with IC/BPS present altered systemic TLR4-mediated inflammatory responses, which were significantly correlated with reported pain severity. In the present study, we sought to determine whether altered TLR4 activation plays a role in pelvic/bladder pain seen in patients with IC/BPS using our validated IC/BPS-like transgenic autoimmune cystitis model (URO-OVA). URO-OVA mice developed responses consistent with pelvic and bladder pain after cystitis induction, which was associated with increased splenocyte production of TLR4-mediated proinflammatory cytokines IL-1β, IL-6, and TNF-α. Increased spinal expression of mRNAs for proinflammatory cytokines IL-6 and TNF-α, glial activation markers CD11b and glial fibrillary acidic protein, and endogenous TLR4 ligand high mobility group box 1 was also observed after cystitis induction. Compared with URO-OVA mice, TLR4-deficient URO-OVA mice developed significantly reduced nociceptive responses, although similar bladder inflammation and voiding dysfunction, after cystitis induction. Intravenous administration of TAK-242 (a TLR4-selective antagonist) significantly attenuated nociceptive responses in cystitis-induced URO-OVA mice, which was associated with reduced splenocyte production of TLR4-mediated IL-1β, IL-6, and TNF-α as well as reduced spinal expression of mRNAs for IL-6, TNF-α, CD11b, glial fibrillary acidic protein, and high mobility group box 1. Our results indicate that altered TLR4 activation plays a critical role in bladder nociception independent of inflammation and voiding dysfunction in the URO-OVA model, providing a potential mechanistic insight and therapeutic target for IC/BPS pain.

Topics: Analgesics; Animals; Autoimmune Diseases; Cells, Cultured; Cystitis, Interstitial; Cytokines; Disease Models, Animal; Inflammation Mediators; Mice, Inbred C57BL; Mice, Knockout; Nociceptive Pain; Ovalbumin; Pain Threshold; Signal Transduction; Spine; Spleen; Sulfonamides; Toll-Like Receptor 4; Urinary Bladder; Urodynamics

Bladder inflammation frequently causes cystitis pain and lower urinary tract dysfunction (LUTD) such as urinary frequency and urgency. Although mast cells have been identified to play a critical role in bladder inflammation and pain, the role of mast cells in cystitis-associated LUTD has not been demonstrated. Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic and debilitating inflammatory condition of the urinary bladder characterized by the hallmark symptoms of pelvic pain and LUTD. In this study we investigated the role of mast cells in LUTD using a transgenic autoimmune cystitis model (URO-OVA) that reproduces many clinical correlates of IC/BPS. URO-OVA mice express the membrane form of the model antigen ovalbumin (OVA) as a self-antigen on the urothelium and develop bladder inflammation upon introduction of OVA-specific T cells. To investigate the role of mast cells, we crossed URO-OVA mice with mast cell-deficient KitW-sh mice to generate URO-OVA/KitW-sh mice that retained urothelial OVA expression but lacked endogenous mast cells. We compared URO-OVA mice with URO-OVA/KitW-sh mice with and without mast cell reconstitution in response to cystitis induction. URO-OVA mice developed profound bladder inflammation with increased mast cell counts and LUTD, including increased total number of voids, decreased mean volume voided per micturition, and decreased maximum volume voided per micturition, after cystitis induction. In contrast, similarly cystitis-induced URO-OVA/KitW-sh mice developed reduced bladder inflammation with no mast cells and LUTD detected. However, after mast cell reconstitution URO-OVA/KitW-sh mice restored the ability to develop bladder inflammation and LUTD following cystitis induction. We further treated URO-OVA mice with cromolyn, a mast cell membrane stabilizer, and found that cromolyn treatment reversed bladder inflammation and LUTD in the animal model. Our results provide direct evidence for the role of mast cells in cystitis-associated LUTD, supporting the use of mast cell inhibitors for treatment of certain forms of IC/BPS.

Topics: Animals; Autoimmune Diseases; Behavior, Animal; CD8-Positive T-Lymphocytes; Cells, Cultured; Cromolyn Sodium; Cystitis, Interstitial; Cytokines; Disease Models, Animal; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Pelvic Pain; Proto-Oncogene Proteins c-kit; RNA, Messenger; Urinary Bladder

To investigate the molecular signature underlying experimental interstitial cystitis (IC) using cDNA microarray.. Microarray gene expression profiles are studied in bladder epithelium of C57BL/6 mice with ovalbumin or substance P-induced experimental IC versus Escherichia coli lipopolysaccharide-induced bacterial cystitis.. Main findings are summarized as follows: firstly, a "75-gene" model was discovered to contain high expressions of bladder epithelium which feature in experimental IC. Secondly, glucose, lipid, nucleotide, xenobiotics, and amino acid metabolisms are involved in. Thirdly, T-cell-mediated immune and inflammatory responses are observed. Fourthly, Wnt, Tgf-beta, Mapk, and insulin growth factor receptor signaling pathways are also involved in. In addition, experimental IC leads to Ephrin- and Semaphorin-mediated axon guidance promoting parasympathetic inflammatory reflexes.. Further characterization of human IC-induced gene expression profiles would enable the use of genome-based expression profiling for the therapeutic targets and diagnosis of IC.

Topics: Animals; Cluster Analysis; Cystitis, Interstitial; Ephrins; Escherichia coli; Gene Expression Profiling; Lipopolysaccharides; Lymphocyte Activation; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Nitric Oxide Synthase Type III; Oligonucleotide Array Sequence Analysis; Ovalbumin; PAX2 Transcription Factor; Semaphorins; Substance P; Transcriptome; Up-Regulation; Wnt Signaling Pathway

Interstitial cystitis (IC) is a debilitating disease that has been adversely affecting the quality of women's lives for many years. The trigger in IC is not entirely known, and a role for the sensory nerves in its pathogenesis has been suggested. In addition to inflammation, increased mast cell numbers in the detrusor muscle have been reported in a subset of IC patients. Experimentally, several lines of evidence support a central role for substance P and neurokinin-1 (NK-1) receptors in cystitis. The availability of mice genetically deficient in neurokinin-1 receptor (NK-1R(-/-)) allows us to directly evaluate the importance of substance P in cystitis. An unexpected finding of this investigation is that NK-1R(-/-) mice present increased numbers of mast cells in the bladder when compared with wild-type control mice. Despite the increase in mast cell numbers, no concomitant inflammation was observed. In addition, bladder instillation of wild-type mice with a sensitizing antigen induces activation of mast cells and an acute inflammatory response characterized by plasma extravasation, edema, and migration of neutrophils. Antigen-sensitized NK-1R(-/-) mice also exhibit bladder mast cell degranulation in response to antigen challenge. However, NK-1R(-/-) mice are protected from inflammation, failing to present bladder inflammatory cell infiltrate or edema in response to antigen challenge. This work presents the first evidence of participation of NK-1 receptors in cystitis and a mandatory participation of these receptors on the chain of events linking mast cell degranulation and inflammation.

Topics: Animals; Cell Degranulation; Cystitis, Interstitial; Dinitrobenzenes; Dinitrophenols; Female; Hypersensitivity, Immediate; Immunization; Mast Cells; Mice; Mice, Knockout; Ovalbumin; Receptors, Neurokinin-1; Substance P; Urinary Bladder