Expression and Function of Chemokines CXCL9-11 in Micturition Pathways in Cyclophosphamide (CYP)-Induced Cystitis and Somatic Sensitivity in Mice

Abstract

Changes in urinary bladder function and somatic sensation may be mediated, in part, by inflammatory changes in the urinary bladder including the expression of chemokines. Male and female C57BL/6 mice were treated with cyclophosphamide (CYP; 75 mg/kg, 200 mg/kg, i.p.) to induce bladder inflammation (4 h, 48 h, chronic). We characterized the expression of CXC chemokines (CXCL9, CXCL10 and CXCL11) in the urinary bladder and determined the effects of blockade of their common receptor, CXCR3, at the level urinary bladder on bladder function and somatic (hindpaw and pelvic) sensation. qRT-PCR and Enzyme-Linked Immunoassays (ELISAs) were used to determine mRNA and protein expression of CXCL9, CXCL10 and CXCL11 in urothelium and detrusor. In urothelium of female mice treated with CYP, CXCL9 and CXCL10 mRNA significantly (p ≤ 0.01) increased with CYP treatment whereas CXC mRNA expression in the detrusor exhibited both increases and decreases in expression with CYP treatment. CXC mRNA expression urothelium and detrusor of male mice was more variable with both significant (p ≤ 0.01) increases and decreases in expression depending on the specific CXC chemokine and CYP treatment. CXCL9 and CXCL10 protein expression was significantly (p ≤ 0.01) increased in the urinary bladder with 4 h CYP treatment in female mice whereas CXC protein expression in the urinary bladder of male mice did not exhibit an overall change in expression. CXCR3 blockade with intravesical instillation of AMG487 (5 mg/kg) significantly (p ≤ 0.01) increased bladder capacity, reduced voiding frequency and reduced non-voiding contractions in female mice treated with CYP (4 h, 48 h). CXCR3 blockade also reduced (p ≤ 0.01) hindpaw and pelvic sensitivity in female mice treated with CYP (4 h, 48 h). CXC chemokines may be novel targets for treating urinary bladder dysfunction and somatic sensitization resulting from urinary bladder inflammation.

Urinary cytokines in women with refractory detrusor overactivity: A longitudinal study of rotating antibiotic versus placebo treatment

“…More recently, the therapeutic properties of agents specifically targeting individual cytokines have been investigated. Blockade of receptors activated by CXCL10 has been shown to increase bladder capacity, reduce bladder contractility and nerve sensitivity in a murine model of interstitial cystitis [ 53 ]. Antibodies developed to block TNF have revolutionised the treatment of inflammatory bowel diseases, Crohn’s disease and ulcerative colitis [ 54 ].…”

Corticotropin-Releasing Hormone from the Pontine Micturition Center Plays an Inhibitory Role in Micturition

“…Cystitis was induced by injection of cyclophosphamide (150 mg/kg, i.p.) in saline as described by others (LaBerge et al, 2006;Bjorling et al, 2007Bjorling et al, , 2011Everaerts et al, 2010;M. Guo et al, 2018); and 48 h later, the voiding patterns were measured again.…”

The Role of PIEZO1 in Urinary Bladder Function and Dysfunction in a Rodent Model of Cyclophosphamide-Induced Cystitis

“…Harvested bladders from naïve (no CYP)/4 h/24 h/chronic (8 d) CYP-treated mice were placed in cold HEPES solution composed of (mM): 134 NaCl, 6 KCI, 10 glucose, 10 HEPES, 1 MgCl 2 , 2 CaCl 2 , 10 glucose and adjusted to pH 7.4 with NaOH. The bladder was cut open along the midline, and pinned to a Sylgard coated dish where the urothelium was separated from the suburothelium (i.e., lamina propria) and the detrusor using fine forceps and iris scissors ( 25 ). To detect Ca 2+ events, the urothelial sheets were loaded with a Ca 2+ sensitive fluorescent dye (10 μM Cal 520; AAT Bioquest, Inc.) for 90 min at 37°C before being placed in a physiological saline solution (PSS) composed of (mM): 119 NaCI, 4.7 KCI, 24 NaHCO 3 , 1.2 KH 2 PO 4 , 2.5 CaCl 2 , 1.2 MgSO 4 , 7 glucose and constantly bubbled with Biological Gas (95% O 2 , 5% CO 2 ) to maintain pH at 7.4.…”