KAVERNOZAL DÜZ KAS PATOFİZYOLOJİSİNDE YENİLİKLER. Dr. Memduh AYDIN Taksim Eğitim ve Araştırma Hastanesi 2. Üroloji Kliniği

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1 KAVERNOZAL DÜZ KAS PATOFİZYOLOJİSİNDE YENİLİKLER Dr. Memduh AYDIN Taksim Eğitim ve Araştırma Hastanesi 2. Üroloji Kliniği

2 2025 için öngörülen ED sıklığı North America 9.1 million Europe 11.9 million Asia 113 million South/Central America and Caribbean 15.6 million Africa 19.3 million Oceania 0.9 million *Worldwide prevalence will increase from 152 million men in 1995 to 322 million men by 2025 Adapted from McKinlay JB. Int J Impot Res. 2000;12(suppl 4):S6-S11.

3 Penis anatomisi:enine Kesit Dorsal nerve Corpora cavernosa Dorsal vein Dorsal artery Tunica albuginea Corpus spongiosum Flaccid Penis Cavernosal artery Urethra Erect Penis National Erectile Dysfunction Foundation. Understanding Erectile Dysfunction

4 NO ve cgmp nin Ereksiyondaki rolleri Sexual stimulation Corpus cavernosum Erection NO NANC Smooth muscle relaxation -- PDE5 inhibitor cgmp=cyclic guanosine monophosphate. GTP=guanosine triphosphate. NANC=nonadrenergic, noncholinergic neurons. NO=nitric oxide. PDE5=phosphodiesterase type 5.

5 NO ve cgmp nin Ereksiyondaki rolleri Cavernous Nerve Endothelial Cell Smooth Muscle Cell NITRIC OXIDE Guanylate cyclase GTP cgmp cgmp-specific protein kinase X PDE5 5 GMP Decreased Ca 2+ Sildenafil ( Viagra ) Vardenafil ( Levitra) Tadalafil ( Cialis ) Smooth muscle relaxation Erection

6 Flaccid State Kavernozal Venöz Arteryal Carson et al., 1999 Textbook of Erectile Dysfunction

7 Düz kasın kasılma aparatının şematik görünümü tropomyosin actin caldesmon calponin? actin binding Loop I SMA/SMB ATP pocket LC 17 (a+b) LC 20 (phosphorylatable) (SM1/SM2) DiSanto ME, J Androl. 2003; 6 (Suppl):S6-S16.

8 Aktin/Myozin Etkileşmesi

9 Aktin/Myozin Hareket şeması

10 Korpus kavernozum düz kas hücrelerinin immunfloresan mikroskobik görünümü α-actin SM Myosin

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12 Effect of Cavernous Neurotomy on Corpus Cavernosum Smooth Muscle: Alteration in Myosin Isoform Composition and the Sphingosine-1-Phosphate/Rho-kinase Signaling Pathway Memduh Aydin, Rowena Chua, Xinhua Zhang, Arnold Melman, Michael E. DiSanto* Department of Urology, Albert Einstein College of Medicine, Bronx, New York Best Poster Award: 2009 American Urological Association Meeting Chicago, USA

13 Nörotominin penis düz kasında myozin alt tiplerine etkisi Relative expression of SM myosin isoforms in the CC 4 weeks after UCN and BCN. * = p < 0.05 UCN increase compared to sham while # = p < 0.05 BCN decrease compared to sham. n = at least 5 individual rats per group.

14 Nörotominin penis düz kasında toplam myozin oluşumuna etkisi

15 Kavernozal sinir harabiyetinin penis düz kasında myozin alt tiplerine etkisi

16 Nörotominin Sphingosine Kinase-2 oluşumuna etkisi

17 PBOO CCSM Sham CCSM SM-B SM-A RT-PCR for SM-A/SM-B PBOO CCSM Sham CCSM Western-blot for Total Myosin Western-blot for SM-B Note: RT-PCR and Western-blot show that SM-B is overexpressed in PBOO CCSM at both the mrna and protein level. Both increases are almost 2 fold. Chang et al., J Urology 2002; 167:

18 125 mm KCl Stimulation 8 Force (g/100mg tissue) Sham CCSM PBOO CCSM Time (minutes) Note: PBOO CCSM generated force faster than sham and the maximum force obtained is about 50% higher. Chang et al., J Urology 2002; 167:

19 Blebbistatin: Selektif Myosin II İnhibitörü

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21 Blebbistatin ile uyarılan penil ereksiyon Zhang et al., Journal of Sexual Medicine 6(10): , 2009,

22 Blebbistatinin mesane ve korpus kavernozum düz kasında yapmış olduğu gevşeme Rat CC Human CC Rat Aorta Rat Bladder Neg Cont Zhang et al., Journal of Sexual Medicine 6(10): , 2009,

23 Blebbistatin ile uyarılan penil ereksiyon ICP/MAP(%) baseline saline DMSO Zhang et al., Journal of Sexual Medicine 6(10): , 2009,

24 Düz kas hücresi içindeki kasılma mekanizması Calcium Sensitization DiSanto ME, Curr Pharm Des. 2005; 11:

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26 Sphingosine-1-Phosphate/Rho-kinase düz kas kasılma yolu Sandhu et al., American Journal of Obstetrics and Gynecology 200(5):576.e1-7, 2009.

27 Sandhu et al., AM J Obst Gynecol. 2009; 200:576 e1-7. S1P Reseptörlerinin etki yolu

28 Hanel P et al., FASEB J. 2007; 21: S1P nin vücutta en çok bulunduğu yerler

29 Rat korpus kavernozumu ve mesanesindeki S1P1, S1P2 ve S1P3 oranları corpus cavernosum bladder Fold change S1P1 S1P2 S1P3 Sandhu et al., Am J Obst Gynecol. 2009; 200:576 e1-7.

30 Kardiovasküler doku mrna ve proteinlerinde S1P reseptör alt tiplerinin tespiti Alewijsne AE et al., Br J Pharmacol. 2004; 143:

31 Penil protez uygulanan diabetik ve non diabetik hastaların korpus kavernozumunda FTY720-P nin neden olduğu kasılma n = 4 n = 7

32 Diabetik ratta korpus kavernozum düz kasının S1P ile uyarılması sonucu oluşan grafik wash wash 600 Rho-kinase Inhibitor S1P 20 µm H µm S1P 15 µm S1P 10 µm S1P 6 µm S1P 3 µmv S1P 1 µm wash wash wash KCl 60 mm Force (mg) 1:06:40 1:15:00 1:23:20 1:31:40 1:40:00

33 Penil protez uygulanan hastaların korpus kavernozum düz kasında S1P ile oluşan kasılma n = 8 n = 11

34 Diyabeti de olan zayıf ratların korpus kavernozum düz kaslarının S1P ile uyarılması

35 The Sphingosine-1-Phosphate Pathway is Upregulated in Response to Partial Urethral Obstruction in Male Rats and Activates RhoA/Rho-kinase Signaling Memduh Aydin, Keith Downing, Guillermo Villegas, Xinhua Zhang, Rowena Chua, Arnold Melman, Michael E. DiSanto* Department of Urology, Albert Einstein College of Medicine Bronx, New York Poster ICS

36 The sphingosine-1-phosphate pathway is upregulated in response to partial urethral obstruction in male rats and activates RhoA/Rhokinase signalling Memduh Aydin, Keith Downing, Guillermo Villegas, Xinhua Zhang, Rowena Chua, Arnold Melman and Michael E. DiSanto Department of Urology, Albert Einstein College of Medicine, Bronx, NY, USA Correspondence to Michael E. DiSanto, Albert Einstein College of Medicine, Room 744, Forchheimer Building, 1300 Morris Park Avenue, Bronx, NY 10461, USA. KEYWORDS bladder outlet obstruction sphingosine-1-phosphate Rho-kinase ABSTRACT To examine the effect of partial urethral obstruction (PUO) on the sphingosine-1-phosphate (S1P, a bioactive lipid shown to modulate smooth muscle, SM) pathway in the bladders of male rats, and to determine the effect of PUO on the RhoA/Rho-kinase (ROK) pathway, and whether there is a molecular cross-talk with the S1P pathways associated with bladder overactivity (S1P1-S1P3, where S1P1 is associated with nitric oxide-mediated SM relaxation, and S1P2 and S1P3 receptors are associated more with SM contraction via the ROK pathway).

37 SPP1/SPP2 No Change RhoA & ROKα ~ 2.0 -fold Similar Protein Pattern S1P (SHAM) S1P (PUO) S1P/H-1152 Aydin M. et al., BJUI 2009

38 Düz kas hücre kasılmasında iyon kanalları Melman et al. Urol Clin North Am, 34:619-30, 2007

39 Maksi-K kanallarının moleküler yapısı Orio P et al., News Physiol Sci 17: , 2002

40 Potasyum kanallarının erektil disfonksiyondaki yeri

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42 SONUÇ RhoA/ROK İnhibitörleri ( Y-27632, H-1152 VB. ) İyon Kanalları agonistleri ve antagonistleri ( özellikle MAXİ K KANAL AKTİVATÖRLERİ ) Sphingolipid agonistleri ve antagonistleri ( JTE, VPC, SURAMİN, FTY VB. ) Myozin inhibitörleri ( BLEBBİSTATİN )