MP49-18 NITRIC OXIDE CONTROLS NGF SECRETION IN BLADDER CELLS IN VITRO THROUGH MATRIX METALLOPROTEINASES
Bibliographic record
Abstract
You have accessJournal of UrologyCME1 May 2022MP49-18 NITRIC OXIDE CONTROLS NGF SECRETION IN BLADDER CELLS IN VITRO THROUGH MATRIX METALLOPROTEINASES Stephanie Sirmakesyan, Aya Hajj, Aalya Hamouda, Philippe Cammisotto, and Lysanne Campeau Stephanie SirmakesyanStephanie Sirmakesyan More articles by this author , Aya HajjAya Hajj More articles by this author , Aalya HamoudaAalya Hamouda More articles by this author , Philippe CammisottoPhilippe Cammisotto More articles by this author , and Lysanne CampeauLysanne Campeau More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002624.18AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Urine of female patients with overactive bladder syndrome (OAB) was found to contain low levels of nerve growth factor (NGF) compared to healthy controls, with stable concentrations of its precursor proNGF. The same samples displayed elevated levels of nitric oxide (NO) that were associated to insulin resistance, a potential causative factor of OAB related to metabolic syndrome. The present study aims to determine if NO is directly linked to the control of NGF secretion from bladder cells. METHODS: Primary cultures of urothelial and smooth muscle cells (SMCs) were grown from collagenase digestion of rat bladders. Expression of NGF, proNGF and MMP-9 mRNA were measured by RT-qPCR. Intracellular MMP-9 was assessed by immunoblotting. Levels of NGF, proNGF and cyclic GMP were measured by ELISA kits. Activities of plasmin and MMP-7 (enzymes converting proNGF into NGF) and MMP-9 (the major protease degrading NGF into peptides) were assessed. RESULTS: Incubation of cells in a hyperglycemic medium (27 mM glucose) led to increased secretion of nitric oxide and in parallel decreased NGF in the culture medium. The NO generator sodium nitroprusside (SNP) (300 µM) incubated for 24 hours also decreased NGF secretion while proNGF levels were stable. SNP potently increased cyclic GMP levels in SMCs, but decreased it in urothelial cells. Stable permeable analogs of cGMP, 8-(4-Chlorophenylthio)-cGMP (3 mM) and N2,2'-O-Dibutyryl-cGMP (1 mM) respectively decreased and increased NGF secretion in SMCs and urothelial cells, mimicking the changes observed in cGMP by SNP. On the other hand, RT-qPCR revealed stable expression of NGF and MMP-9 mRNA in SMCs, while they were increased in urothelial cells. In Crispr-Cas9 transfected SMCs, MMP-9 expression was abolished and decrease in NGF secretion by SNP was unaffected demonstrating that SNP effect in SMCs is not mediated by MMP-9. On the other hand, SNP decreases MMP-7 activity and did not affect plasmin levels, showing a decrease of NGF synthesis from proNGF. In urothelial cells, Crispr-cas9 potently inhibited the effect of SNP on NGF secretion while MMP-7 levels were enhanced, suggesting increase in NGF proteolysis and a compensatory mechanism to increase NGF. CONCLUSIONS: Nitric oxide decreases NGF secretion by cells of the bladder through cyclic GMP and matrix metalloproteinases 7 and 9. This control is regulated differently between urothelial and SMCs. Our findings are in concordance with our previous studies of proNGF and NGF in the urine of OAB patients, and link the secretion of NGF to hyperglycaemia/insulin resistance and nitric oxide levels. Source of Funding: Canadian Urology Association © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e861 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Stephanie Sirmakesyan More articles by this author Aya Hajj More articles by this author Aalya Hamouda More articles by this author Philippe Cammisotto More articles by this author Lysanne Campeau More articles by this author Expand All Advertisement PDF downloadLoading ...
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.004 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".