MP89-16 INCREASE IN TNF-ALPHA LEADS TO APOPTOSIS OF NITRERGIC NEURONS IN MAJOR PELVIC GANGLION AFTER BILATERAL CAVERNOUS NERVE INJURY

Abstract

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology II1 Apr 2016MP89-16 INCREASE IN TNF-ALPHA LEADS TO APOPTOSIS OF NITRERGIC NEURONS IN MAJOR PELVIC GANGLION AFTER BILATERAL CAVERNOUS NERVE INJURY Hotaka Matsui, Nikolai A. Sopko, Johanna L. Hannan, Allison A. Reinhardt, Emmanuel Weyne, Max Kates, Xiaopu Liu, Maarten Albersen, and Trinity J. Bivalacqua Hotaka MatsuiHotaka Matsui More articles by this author , Nikolai A. SopkoNikolai A. Sopko More articles by this author , Johanna L. HannanJohanna L. Hannan More articles by this author , Allison A. ReinhardtAllison A. Reinhardt More articles by this author , Emmanuel WeyneEmmanuel Weyne More articles by this author , Max KatesMax Kates More articles by this author , Xiaopu LiuXiaopu Liu More articles by this author , Maarten AlbersenMaarten Albersen More articles by this author , and Trinity J. BivalacquaTrinity J. Bivalacqua More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2477AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Erectile dysfunction (ED) is induced by inflammation in cavernous nerve (CN) after radical prostatectomy (RP). This study examines changes of inflammatory cytokine: tumor necrosis factor-alpha (TNF-α) after bilateral CN injury (BCNI) in vivo and its effect on neurite outgrowth from major pelvic ganglion (MPG) ex vivo. METHODS Male rats underwent BCNI or sham surgery. MPGs were harvested 2, 7 and 14 days after surgery (n=10/group). MPGs were processed for qPCR, Western blot (WB), and immunofluorescence (IF) for TNF-α. MPGs were also harvested from untouched rats and cultured in Matrigel with exogenous TNF-α (eTNF-α) in concentrations of 0, 10, 20, 30 ng/mL (n=5/group). Neurite lengths were measured 3 days after culture. MPGs were then processed for qPCR to evaluate gene expression of neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase (TH), and caspase (CASP)-1 (n=5/group). Additional MPGs were cultured in Matrigel with/without eTNF-α 20 ng/mL to perform IF for TH and nNOS and to evaluate protein amount of CASP3 by WB (n=3/group). RESULTS BCNI increased gene and protein expression of TNF-α (p<0.05). Maximal increase was at 2 days for gene and at 7 days for protein expression of TNF-α. IF demonstrated that TNF-α mainly localized in periphery at 2 days, cytoplasm of soma at 7 days, and interstitial space at 14 days after BCNI. eTNF-α inhibited neurite outgrowth from MPG in a dose-dependent fashion (neurite lengths: 586±20.5, 455±11.5, 408±28.7, 361±2.65 μm; eTNF-α 0, 10, 20, 30 ng/mL, respectively: p<0.01). MPGs cultured with eTNF-α showed increased gene expression of CASP1 (p=0.03) and decreased gene expression of nNOS (p<0.05). Gene expression of TH remained unchanged. WB demonstrated increased protein amount of total CASP3 in MPG cultured with eTNF-α (p=0.03). However, there was no significant difference in ratio of cleaved and total CASP3. IF demonstrated that nNOS+ neurites were shorter than TH+ neurites in MPGs cultured with eTNF-α (p<0.01), while there was no difference in neurite lengths between nNOS+ neurites and TH+ neurites in control (p=0.29). CONCLUSIONS This study demonstrates that TNF-α is increased after BCNI and impairs neurite outgrowth from MPG in a dose-dependent fashion. Furthermore, eTNF-α upregulated gene and protein expression of apoptosis markers and downregluated gene and protein expression of nNOS, suggesting that TNF-α selectively induces apoptosis of nitrergic neurons. These results suggest that TNF-α antagonists may prevent post-RP ED. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1142-e1143 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Hotaka Matsui More articles by this author Nikolai A. Sopko More articles by this author Johanna L. Hannan More articles by this author Allison A. Reinhardt More articles by this author Emmanuel Weyne More articles by this author Max Kates More articles by this author Xiaopu Liu More articles by this author Maarten Albersen More articles by this author Trinity J. Bivalacqua More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...