Proteomic Investigation on Chronic Bladder Irritation in the Rat

Tyagi, Pradeep; Chen, Xuan; Hayashi, Yukio; Yoshimura, Naoki; Chancellor, Michael B.; Miguel, Fernando de

doi:10.1016/j.urology.2007.10.069

Cited by 10 publications

(8 citation statements)

References 19 publications

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“…In order to check the specificity, we performed two-dimensional Western blot analysis using the rat aorta. As shown in figure 1, clone 75–7 recognized a single spot of 22 kDa protein at around pI 9.0, which is consistent with previously published data [25]. …”

Section: Resultssupporting

confidence: 92%

Quantitative Histological Analysis of SM22α (Transgelin) in an Adriamycin-Induced Focal Segmental Glomerulosclerosis Model

Wang

Sakatsume²,

Sakamaki³

et al. 2011

Nephron Exp Nephrol

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Background/Aims: SM22α, transgelin, has been revealed to be specifically expressed in glomerular epithelial cells and interstitial cells, according to the nature of the renal injury. In this study, quantitative analyses of SM22α positivity were performed to investigate the pathological significance of its expression. Methods: Kidney samples of adriamycin nephropathy underwent immunohistochemistry with a newly established anti-SM22α monoclonal antibody. The SM22α positivity was quantified by an image analyzer. The correlation of the histological values with biochemical data was investigated statistically. Microstructural localization of SM22α was studied by immunoelectron microscopy. Results: SM22α was expressed along the dense basal microfilaments of degenerating podocytes, and diffusely in interstitial cells. Both the extent and intensity of SM22α expression in glomerular and tubulointerstitial area were correlated with the deterioration of renal function and the severity of proteinuria. Stepwise multiple linear regression analysis revealed that the extent of its positivity in glomerular or tubulointerstitial area was the determinant of the amount of proteinuria or the deterioration of creatinine clearance (Ccr), respectively. Inversely, the deterioration of Ccr was the most important predictor of SM22α expression. Conclusion: SM22α expression in podocytes and interstitial cells represented the severity of proteinuria and the deterioration of renal function. SM22α expression in renal tissues might be a hallmark of kidney diseases.

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Section: Resultssupporting

confidence: 92%

Quantitative Histological Analysis of SM22α (Transgelin) in an Adriamycin-Induced Focal Segmental Glomerulosclerosis Model

Wang

Sakatsume²,

Sakamaki³

et al. 2011

Nephron Exp Nephrol

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“…However, the predominant expression of S100A8-S100A9 by uroepithelial cells was an unexpected finding. Typically, these cells do not express this heterodimer complex, and its presence in the uroepithelium is normally considered to be a biomarker of immune dysregulation or neoplastic transformation (29,30). We did not detect S100A8-S100A9 expression in the uroepithelia of animals from 48 and 72 h p.i., but neutrophils were positive for S100A8-S100A9, which confirmed the efficacy of the assay.…”

Section: Discussionmentioning

confidence: 55%

Complicated Urinary Tract Infection Is Associated with Uroepithelial Expression of Proinflammatory Protein S100A8

et al. 2009

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F344 rats chronically infected with Ureaplasma parvum develop two distinct profiles: asymptomatic urinary tract infection (UTI) and UTI complicated by struvite urolithiasis. To identify factors that affect disease outcome, we characterized the temporal host immune response during infection by histopathologic analysis and in situ localization of U. parvum. We also used differential quantitative proteomics to identify distinguishing host cellular responses associated with complicated UTI. In animals in which microbial colonization was limited to the mucosal surface, inflammation was indistinguishable from that which occurred in shaminoculated controls, and the inflammation resolved by 72 h postinoculation (p.i.) in both groups. However, inflammation persisted in animals with microbial colonization that extended into the deeper layers of the submucosa. Proteome profiling showed that bladder tissues from animals with complicated UTIs had significant increases (P < 0.01) in proteins involved in apoptosis, oxidative stress, and inflammation. Animals with complicated UTIs (2 weeks p.i.) had the highest concentrations of the proinflammatory protein S100A8 (P < 0.005) in bladder tissues, and the levels of S100A8 positively correlated with those of proinflammatory cytokines GRO/KC (P < 0.003) and interleukin-1␣ (P < 0.03) in urine. The bladder uroepithelium was a prominent cell source of S100A8-S100A9 in animals with complicated UTIs (2 weeks p.i.), which was not detected in animals with asymptomatic UTIs (2 weeks p.i.) or in any bladder tissues harvested at earlier p.i. time points. Based on these results, we surmise that invasive colonization of the bladder triggers chronic inflammation and immune dysregulation, which may be critical to struvite formation.Struvite or infection stones form as a result of complicated urinary tract infections (UTI) caused by urease-producing bacteria such as Proteus, Klebsiella, Serratia, and Ureaplasma species (5,8,9,17). Bacterial urease breaks down urea into ammonia, resulting in urine's becoming supersaturated with ammonia. When this occurs, the urine pH rises and the solubility of magnesium ammonium phosphate decreases, leading to crystal deposition and stone formation. Infection is always an underlying cause of struvite urolithiasis (5, 9). Therefore, it is not surprising that known predisposing factors such as renal tubular acidosis, neurogenic bladder, urinary tract obstructions, and chronic use of indwelling catheters (5, 9) are related to elements that increase patient susceptibility to UTI. Although infections usually induce an inflammatory response in the host, to our knowledge, the role of the host immune response as a potential predisposing factor in struvite urolithiasis has not been evaluated or considered to be important in disease pathogenesis. We present data suggesting that the host immune response plays a critical role in the development of struvite stones and that the proinflammatory protein S100A8-S100A9 may play a critical role in the chronic inflammation and immune...

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“…14 Also, investigation of bladder outlet obstruction in rats showed that downregulation of transgelin might decrease bladder contractility, 15 and the study of chronic bladder irritation in rats induced by HCl drew the same conclusion. 16 The actin-binding protein, transgelin, might play a role in contractility in smooth muscle by affecting actin filament organization.…”

Section: Discussionmentioning

confidence: 99%

Effects of long‐term ketamine administration on rat bladder protein levels: A proteomic investigation using two‐dimensional difference gel electrophoresis system

Huang

Shan

et al. 2013

Int J of Urology

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Abbreviations & Acronyms 2D-DIGE = two-dimensional difference gel electrophoresis CHAPS = 3- dimethylammonio] propanesulfonate Cy = cyanine dye DIGE = difference gel electrophoresis DTT = DL-Dithiothreitol GAPDH = glyceraldehyde 3-phosphate dehydrogenase HK = high dose of ketamine IC = interstitial cystitis IEF = isoelectric focusing LK = low dose of ketamine MALDI-TOF-MS = matrixassisted laser desorption/ionization time-of-light mass spectrometry NS = normal saline SDS-PAGE = sodium dodecyl sulphate-polyacrylamide gel electrophoresis Objectives: Long-term ketamine abuse can affect the urinary system, resulting in interstitial cystitis-like syndrome. However, its pathogenesis remains unclear. In the present study, a proteomic approach of two-dimensional difference gel electrophoresis followed by matrix-assisted laser desorption/ionization time-of-light mass spectrometry was carried out to investigate the potential disease-associated proteins in a rat model of ketamine-associated cystitis. Methods: Rats were randomly assigned to control, normal saline, low dose of ketamine (10 mg/kg) and high-dose of ketamine (50 mg/kg) groups with six rats in each group. The two experimental groups were given ketamine hydrochloride i.p. daily, whereas the normal saline group rats were treated with saline. After 16 weeks of treatment, all bladders were excised, and samples from normal saline and high dose of ketamine groups were resolved in two-dimensional difference gel electrophoresis. Differentially expressed spots were excised and identified by matrix-assisted laser desorption/ionization time-of-light mass spectrometry. Phosphoprotein and nonphosphoprotein purification, histopathology, immunohistochemistry, and western blot were carried out in all groups. Results: Histological study showed hyperplastic epithelium and inflammatory cells infiltration in the high dose of ketamine-treated rat bladders. Two-dimensional difference gel electrophoresis revealed 30 altered expressions between the normal saline and high dose of ketamine-treated group. Among these proteins, two upregulated and two downregulated protein spots were all identified as smooth muscle protein-22/transgelin. Immunohistochemical staining and western blot analysis showed that the expression of total transgelin had no significant difference between groups. However, the expression of phosphorylated transgelin in the low-dose and high dose of ketamine groups was increased, whereas the non-phosphorylated transgelin was decreased when compared with the normal saline group. Conclusions: Long-term ketamine abuse induces phosphorylation of transgelin in the bladder wall, and this might play an important role in the pathogenesis of ketamineassociated cystitis.

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Proteomic Investigation on Chronic Bladder Irritation in the Rat

Cited by 10 publications

References 19 publications

Quantitative Histological Analysis of SM22α (Transgelin) in an Adriamycin-Induced Focal Segmental Glomerulosclerosis Model

Quantitative Histological Analysis of SM22α (Transgelin) in an Adriamycin-Induced Focal Segmental Glomerulosclerosis Model

Complicated Urinary Tract Infection Is Associated with Uroepithelial Expression of Proinflammatory Protein S100A8

Effects of long‐term ketamine administration on rat bladder protein levels: A proteomic investigation using two‐dimensional difference gel electrophoresis system

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