Effect of Glucagon-Like Peptide-1 Receptor Agonist Liraglutide on Rat Detrusor Muscle Contraction Response

Öz

Aim: Glucagon-like peptide-1 (GLP-1) is an incretin hormone synthesized from proglucagon in intestinal L cells. GLP-1 analogs, which are widely used in the treatment of diabetes mellitus, are reported to be effective in neuroprotective, procognitive and prevention of diabetes-related complications by providing neuroplasticity as well as antidiabetic effects. Urinary dysfunctions are frequently observed in diabetes and obesity. Determination of the effects of GLP-1 receptor agonists directly on bladder smooth muscle, independent of their blood glucose lowering effect, may reveal beneficial effects in preventing the development of overactive bladder in these patient groups. The aim of this study was to investigate the effect of GLP-1 receptor agonist, which has been reported to have regulatory effects on gastrointestinal smooth muscle functions, on bladder smooth muscle contraction and relaxation responses. Material and Methods: Male adult Wistar Albino rats weighing 300-325 g were used in the study (n=7). An organ bath system was used to examine the contraction and relaxation responses of detrusor muscle strips. Muscle strips with intact mucosa weres uspended in an isolated organ bath and isometric contraction response was induced with carbachol (3×10-6 M). Liraglutide as GLP-1 receptor agonist was applied cumulatively (10-8-10-2 M) to carbachol-contracted smooth muscle strips and the change in smooth muscle contraction responses was determined. Isoproterenol (10-8-10-2 M) was used to examine relaxation responses. Relaxation responses were recorded by cumulatively applying liraglutide to strips of smooth muscle previously contracted with carbachol. In addition, in order to determine the mechanism of action of GLP-1, neuronal nitric oxide synthase (nNOS) inhibitör Nω-Nitro-L-arginine (L-NNA) (10 mM) was added to the organ bath and its effect on relaxation responses was evaluated. Statistical evaluation of the results was performed using Mann- Whitney U and Kruskal Wallis tests. Results: The GLP-1 receptor agonist liraglutide did not cause a statistically significant change in the contractile response to carbachol at a dose of 3×10-6 M (p=0.768). Liraglutide (at doses of 10-4 and 10-2 M) applied cumulatively to smooth muscle strips previously contracted with 3×10-6 M carbachol caused a similar relaxation response as isoprotrenol. Addition of LNNA to the isolated organ bath medium caused a statistically significant decrease in the liraglutide-dependent relaxation response (p=0.019, p=0.033). Conclusion: The results of the study show that GLP-1 can cause relaxation of the bladder smooth muscle wall, most likely through the GLP-1 receptor. Data obtained in vitro and from healthy detrusor muscle tissue suggest that nitric oxide plays a role in the mechanisms of the relaxation response induced by liraglutide. These results need to be supported in further studies in overactive bladder models and in vivo conditions.

Anahtar Kelimeler

• GLP-1
• bladder
• contractile response
• nitric oxide

Glukagon Benzeri Peptit -1 Reseptör Agonisti Liraglutidin Sıçan Detrüsor Kas Kasılma Yanıtı Üzerine Etkisi

Öz

Amaç: Glukagon benzeri peptid-1 (GLP-1), bağırsak L hücrelerinde proglukagondan sentezlenen bir inkretin hormondur. Diyabetes mellitus tedavisinde yaygın olarak kullanılan GLP-1 analoglarının antidiyabetik etkilerinin yanı sıra nöroplastisiteyi sağlayarak nöroprotektif, prokognitif ve diyabete bağlı komplikasyonların önlenmesinde etkili olduğu bildirilmektedir. Üriner disfonksiyonlar diyabet ve obezitede sıklıkla gözlenmektedir. Kan şekerini düşürücü etkisinden bağımsız olarak GLP-1 reseptör agonistlerinin direkt mesane düz kasının üzerindeki etkilerinin belirlenmesi bu hasta gruplarında aşırı aktif mesane gelişiminin önlenmesinde yararlı etkiler ortaya çıkarabilir. Bu çalışmanın amacı gastrointestinal düz kas fonksiyonları üzerine düzenleyici etkileri bildirilen GLP-1 reseptör agonistinin mesane düz kas kasılma ve gevşeme yanıtları üzerine olan etkisini incelemektir. Gereç ve Yöntemler: Çalışmada 300-325 g ağırlığında erkek yetişkin Wistar Albino cinsi sıçanlar kullanılmıştır (n=7). Detrüsör kas şeritlerinin kasılma ve gevşeme yanıtlarını incelemek için organ banyosu sistemi kullanılmıştır. Mukozası sağlam kas şeritleri izole organ banyosuna asılarak izometrik kasılma yanıtı karbakol ( 3×10-6 M) ile oluşturulmuştur. Karbakolle kasılmış düz kas şeritlerine GLP-1 reseptör agonisti olarak liraglutid kümülatif (10-8-10-2 M) olarak uygulanarak düz kas kasılma yanıtlarındaki değişim belirlenmiştir. Gevşeme yanıtları incelemek için izoproterenol (10-8-10-2 M) kullanılmıştır. Karbakolle kasılmış düz kas şeritlerine kümülatif olarak liraglutid uygulanarak gevşeme yanıtları kaydedilmiştir. Ayrıca GLP-1’in etki mekanizmasını tespit edebilmek için organ banyosuna nöronal nitrik oksit sentaz (nNOS) inhibitörü Nω-Nitro-L-arginine (L-NNA) (10 mM) eklenerek gevşeme yanıtları üzerine etkisi değerlendirilmiştir. Sonuçların istatistiksel değerlendirmesi Mann-Whitney U ve Kruskal Wallis testleri kullanılarak yapılmıştır. Bulgular: GLP-1 reseptör agonisti liraglutid 3×10-6 M dozunda karbakolle oluşan kasılma yanıtında istatistiksel olarak anlamlı bir değişime neden olmamıştır (p=0,768). Önceden 3×10-6 M karbakol ile kasılmış düz kas şeritlerine kümülatif olarak uygulana liraglutid (10-4 ve 10-2 M dozlarında) izoprotrenol ile benzer gevşeme yanıtına neden olmuştur. İzole organ banyosu ortamına LNNA eklenmesi, liraglutid bağımlı gevşeme yanıtında istatistiksel olarak anlamlı azalmaya neden olmuştur (p=0,019, p=0,033). Sonuç: Çalışmanın sonuçları GLP-1’in büyük olasılıkla mesane düz kas duvarında GLP-1 reseptörü aracılığıyla gevşemeye neden olabildiğini göstermektedir. İn vitro koşullarda ve sağlıklı detrüsör kas dokusundan elde edilen veriler, liraglutid ile oluşan gevşeme yanıtının mekanizmaları arasında nitrik oksitin rol oynadığını düşündürmektedir. Bu sonuçların aşırı aktif mesane modellerinde ve in vivo koşullarda daha ileri çalışmalarda desteklenmesi gerekmektedir.

Anahtar Kelimeler

• GLP-1
• mesane
• kasılma yanıtı
• nitrik oksit

Kaynakça

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