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43 BAY 41-2272 inhibited migration of smooth muscle cells from rat corpus cavernosum tissue for study of erectile dysfunction
Title
43
BAY 41-2272 inhibited migration of smooth muscle cells from rat corpus cavernosum tissue for study of erectile dysfunction Eur Urol Suppl 2015;14/2;e43
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Kim H.S. 1 , Chung H.1 , Sul C.K. 2 , Song K.H. 2 1 Konkuk
University School of Medicine, Dept. of Urology, Chungju, South Korea, 2 Chungnam National University School of Medicine, Dept.
of Urology, Daejeon, South Korea INTRODUCTION & OBJECTIVES: Smooth muscle cells (SMCs) migration and proliferation are key components of fibrosis and inflammation that lead to erectile dysfunction (ED). Bay 41-2272 is an introduced orally available agent that that directly stimulates soluble guanylate cyclase (sGC) and sensitizes it to its physiological stimulator, nitric oxide. In our study, we applied a novel technique for the isolation of the rat cavernous SMCs and investigated the role of sildenafil in SMCs. And we studied the smooth muscle cell migrations of BAY 41-2272 in a rat model of cavernous nerve injury (CNI) and compared it to sildenafil. MATERIAL & METHODS: Using 8-weeks-old male Sprague Dawley rat, sham and CNI operation under microscope were performed. After 2 weeks, penile tissues were harvested. We applied the two stage tissue culture methods utilizing Matrigel-based sprouting culture system to facilitate stromal cell sprouting and adherent culture system using D-valine to eliminate contamination of fibroblasts from SMCs. The expression of α-SMA, desmin, PECAM-1, and S100A4 in the SMCs was determined by standard immunofluorescent staining and immunoblotting. Flow cytometry analysis (FCA) was performed. SMCs migration with/without sildenafil and Bay 41-2272 was measured using a Boyden chamber. RESULTS: ICP/MAP ratio of sham was significantly increased compared with CNI group. Immunocytochemical staining in the SMCs showed greater α-SMA and desmin-positive and PCAM-1 and S100A4-negative fluorescence. Moreover, whereas the expression of α-SMA and desmin was detected in the SMCs, that of PECAM-1 and S100A4 was not. Using FCA analysis, rat penile SMCs made up 69.9–79.3% of the cells analyzed and fibroblasts 5.2%. Migration of penile SMCs from the sham-operated group was increased by TGF- β treatment compared with cells from the CNI group and was decreased by sildenafil treatment in both groups. Additionally, it was decreased more than Bay 41-2272 and sildenafil compared with sildenafil only.
file:///S|/IM/EURSUP/2015%20EAU%20Abstracts/content/data/43.html[19/02/2015 08:13:15]
Title
Figure. Effects of sildenafil on TGF-β-induced migration of rat penile SMCs from the sham-operated control and BCNC (bilateral cavernous nerve crushing injury) groups. CONCLUSIONS: Our results showed that SMCs migration in rat penis was increased by TGF-β and it was inhibited by sildenafil and Bay 41-2272. Bay 41-2272, directly stimulates sGC, enhanced sildenafil effect in the penile SMCs. We suggested that the increase of penile SMCs migration could be related with ED and sildenafil combined with Bay 41-2272 should be improved SMCs function in CNI.
file:///S|/IM/EURSUP/2015%20EAU%20Abstracts/content/data/43.html[19/02/2015 08:13:15]
43
BAY 41-2272 inhibited migration of smooth muscle cells from rat corpus cavernosum tissue for study of erectile dysfunction Eur Urol Suppl 2015;14/2;e43
Print! Print!
Kim H.S. 1 , Chung H.1 , Sul C.K. 2 , Song K.H. 2 1 Konkuk
University School of Medicine, Dept. of Urology, Chungju, South Korea, 2 Chungnam National University School of Medicine, Dept.
of Urology, Daejeon, South Korea INTRODUCTION & OBJECTIVES: Smooth muscle cells (SMCs) migration and proliferation are key components of fibrosis and inflammation that lead to erectile dysfunction (ED). Bay 41-2272 is an introduced orally available agent that that directly stimulates soluble guanylate cyclase (sGC) and sensitizes it to its physiological stimulator, nitric oxide. In our study, we applied a novel technique for the isolation of the rat cavernous SMCs and investigated the role of sildenafil in SMCs. And we studied the smooth muscle cell migrations of BAY 41-2272 in a rat model of cavernous nerve injury (CNI) and compared it to sildenafil. MATERIAL & METHODS: Using 8-weeks-old male Sprague Dawley rat, sham and CNI operation under microscope were performed. After 2 weeks, penile tissues were harvested. We applied the two stage tissue culture methods utilizing Matrigel-based sprouting culture system to facilitate stromal cell sprouting and adherent culture system using D-valine to eliminate contamination of fibroblasts from SMCs. The expression of α-SMA, desmin, PECAM-1, and S100A4 in the SMCs was determined by standard immunofluorescent staining and immunoblotting. Flow cytometry analysis (FCA) was performed. SMCs migration with/without sildenafil and Bay 41-2272 was measured using a Boyden chamber. RESULTS: ICP/MAP ratio of sham was significantly increased compared with CNI group. Immunocytochemical staining in the SMCs showed greater α-SMA and desmin-positive and PCAM-1 and S100A4-negative fluorescence. Moreover, whereas the expression of α-SMA and desmin was detected in the SMCs, that of PECAM-1 and S100A4 was not. Using FCA analysis, rat penile SMCs made up 69.9–79.3% of the cells analyzed and fibroblasts 5.2%. Migration of penile SMCs from the sham-operated group was increased by TGF- β treatment compared with cells from the CNI group and was decreased by sildenafil treatment in both groups. Additionally, it was decreased more than Bay 41-2272 and sildenafil compared with sildenafil only.
file:///S|/IM/EURSUP/2015%20EAU%20Abstracts/content/data/43.html[19/02/2015 08:13:15]
Title
Figure. Effects of sildenafil on TGF-β-induced migration of rat penile SMCs from the sham-operated control and BCNC (bilateral cavernous nerve crushing injury) groups. CONCLUSIONS: Our results showed that SMCs migration in rat penis was increased by TGF-β and it was inhibited by sildenafil and Bay 41-2272. Bay 41-2272, directly stimulates sGC, enhanced sildenafil effect in the penile SMCs. We suggested that the increase of penile SMCs migration could be related with ED and sildenafil combined with Bay 41-2272 should be improved SMCs function in CNI.
file:///S|/IM/EURSUP/2015%20EAU%20Abstracts/content/data/43.html[19/02/2015 08:13:15]