ABSTRACT: What's known on the subject? and What does the study add? In the urinary bladder, histological studies suggest a network of functionally connected interstitial cells of Cajal (ICCs) are located between the urothelium and sensory nerve endings, which might transfer signals directly between them. Purinergic P2X(3) receptors may play roles in the micturition reflex pathway, and its expression profiles in ICCs could be altered in urinary bladder dysfunction. The present experiments showed a novel time-dependent P2X(3) receptor up-regulation in ICCs in an experimental rat model of partial bladder outlet obstruction. OBJECTIVE: • To investigate the expression and electrophysiological characteristics of purinergic P2X(3) receptors in interstitial cells of Cajal (ICCs) at different time points after partial bladder outlet obstruction (PBOO) in rats. MATERIALS AND METHODS: • In all, 48 female Sprague-Dawley rats were randomly divided into four groups: 4, 6 and 8 week PBOO groups and sham-operated controls. At 4 weeks after surgery, cystometry was performed to investigate bladder function in vivo. Subsequently, the rats were humanely killed at 4, 6 or 8 weeks and the bladders were harvested for measurements. • P2X(3) expression in ICCs of bladder was investigated by immunofluorescent staining. • The level of P2X(3) mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). • Inward currents in corresponding ICCs after PBOO were investigated electrophysiologically. RESULTS: • Cystometrography showed a valid increase in maximum detrusor pressure in rats subjected to PBOO. The bladder weight in the PBOO group was significantly higher than that in the control group. • In contrast to sham rats, there was a significant increase in the intensity of P2X(3) staining in the ICCs in all PBOO groups. C-kit labelled isolated ICCs were strongly stained with the P2X(3) antibody. • RT-PCR showed that the expression of P2X(3) mRNA was significantly up-regulated at 4, 6 and 8 weeks in the ICCs from the PBOO rats. • In the ICCs, the mean peak amplitude of inward currents was significantly increased in the PBOO groups compared with the control group. CONCLUSIONS: • The expression of P2X(3) receptors showed a time dependent up-regulation in the ICCs of the bladder in rats with PBOO. • PBOO induced the potentiation of P2X(3) receptors function, as evidenced by α, β-methylene ATP-enhanced inward currents in the ICCs of the rat bladder.
BJU International 08/2012; · 2.84 Impact Factor
ABSTRACT: To investigate the expression of hyperpolarization-activated cyclic nucleotide-gated (HCN) protein in the interstitial cells of Cajal (ICCs) of the human bladder and to determine the relative expression of the HCN subtypes.
A total of 30 bladder specimens were obtained, and each bladder sample was divided into 2 parts. Part I, which included mucosa, submucosa, and muscle, was used for immunofluorescence study. Part II, in which the mucosa and submucosa were removed and fresh bladder muscle tissue was kept, was used for Western blotting. First, the immunoreactivity of HCN1, HCN2, HCN3, and HCN4 isoforms was detected in the ICCs of lamina propria using immunofluorescence. Second, the protein expression of HCN1, HCN2, HCN3, and HCN4 isoforms in ICCs of bladder detrusor muscle was analyzed using Western blotting.
Immunofluorescence showed that there were many vimentin- and HCN4-positive ICCs in the lamina propria region and detrusor. Also, only a few c-kit and HCN4-positive ICCs were detected in the lamina propria region and detrusor. Additionally, novel c-kit negative, but HCN4-positive mast cells were found in the lamina propria. Immunofluorescence and Western blotting revealed that HCN1, HCN2, HCN3, and HCN4 channels were all present in the ICCs of the human bladder; however, HCN4 channel expression in the ICCs was greater than in HCN1, HCN2, or HCN3.
HCN1, HCN2, HCN3, and HCN4 channels are identified in ICCs of human bladder tissue, and the expression of the HCN4 channel exceeded that of the other subtypes.
Urology 07/2012; 80(1):224.e13-8. · 2.43 Impact Factor
ABSTRACT: Renal ischemia-reperfusion (I/R) injury, as a common and clinically important problem, starts with direct damage caused by chemokines and inflammatory cytokines, which is aggravated by specific and nonspecific immune reactions. Recently, IL-17A has been considered to be in a uniquely powerful position between adaptive and innate immunity. The present study investigated the role of IL-17A in renal I/R injury in mice.
We measured the time-course of changes in plasma and renal IL-17A levels using a murine model of renal I/R injury. Then, the protective effect of monoclonal anti-IL-17A antibody, given intravenously at 30 min before or after renal I/R operation, on renal I/R injury was investigated. In addition, the levels of plasma and renal pro- and anti-inflammatory cytokines and chemokines were assessed.
IL-17A was significantly increased in plasma and kidneys after renal I/R injury in mice. Furthermore, intravenous administration of neutralizing monoclonal anti-IL-17A antibody attenuated renal I/R injury by evaluating renal function and histopathology. In addition, administration of anti-IL-17A antibody substantially reduced the plasma and renal levels of many pro-inflammatory mediators (TNF-α, IL-6, high-mobility group box 1 (HMGB1), IL-1β, IL-17A, macrophage inflammatory protein-1α (MIP-1α), and monocyte chemoattractant protein-1 (MCP-1), as well as increased the plasma and renal levels of anti-inflammatory cytokines IL-10 and transforming growth factor β (TGF-β).
The above data suggest that IL-17A has a detrimental effect on renal I/R injury via facilitating the production of pro-inflammatory cytokines and chemokines as well as hampering the production of anti-inflammatory cytokines.
Journal of Surgical Research 11/2011; 171(1):266-74. · 2.25 Impact Factor