MP49-18 NITRIC OXIDE CONTROLS NGF SECRETION IN BLADDER CELLS IN VITRO THROUGH MATRIX METALLOPROTEINASES
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Résumé
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 ...
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,004 | 0,001 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».