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017 Apelin-13 Protects Corpus Cavernosum Against Hypercholesterolemia-Induced Fibrosis
S8
WMSM 2016 Abstracts
(Allo), hCt+allogenic-PBMCs+cyclosporineA (CsA). Additional hCt were cultured without PBMCs for 24 hours in the presence of media alone, cyclosporineA, or FK506. Tissues were evaluated by IHC and live confocal fluorescent imaging. Myography was used to examine nerve-mediated contraction and relaxation in response to electrical field stimulation. PBMC activation was assessed by flow cytometry and qPCR-array. This study was approved by the IRB. Results: Microscopy demonstrated increased caspase-3/7 activation and TUNEL staining in tissues exposed to allogeneic PBMCs, which was prevented by CsA. Flow cytometry and qPCR-array demonstrated PBMC activation when exposed to allogenic hCt, which was prevented by CsA treatment. Myography demonstrated impaired contraction and relaxation in Allo compared to Auto. CsA treated Allo had similar contraction and surprisingly impaired relaxation compared to non-treated Allo tissues. Compared to media and FK506, CsA impaired nervemediated relaxation. Conclusions: This model may be used to investigate the pathogenesis of rejection and to optimize immunosuppression for penile transplantation. Disclosure: Work supported by industry: no.
content. Accordingly, Apelin-13 enhanced total matrix metalloproteinase (MMP) activity in the mice penis, which was associated with an increased protein expression of MMP-1, MMP-3, MMP-8 and MMP-9, while tissue inhibitor of metalloproteinase(TIMP)-1, TIMP-2 and TIMP-4 were unaltered. These beneficial actions were not associated with changes in nNOS or eNOS protein expression, intracavernosal ROS content, or atherosclerotic plaque deposition. In mouse fibroblast, Apelin-13 inhibited TGFb1-induced fibroblast differentiation into myofibroblast and collagen production. Conclusion: These results suggest a local modulation of Apelin/ APJ system within the corpus cavernosum. Remarkably, Apelin13 attenuated hypercholesterolemic-induced cavernosal fibrosis by: 1) enhancing MMPs expression and activity; 2) inhibiting fibroblast differentiation into myofibroblast; and 3) diminishing collagen production. These results provide the first evidence of Apelin/APJ role on erectile tissue structure/remodeling, and point out this system as a novel potential target to alleviate intracavernosal fibrosis-related disorders. Disclosure: Work supported by industry: no.
017
018
APELIN-13 PROTECTS CORPUS CAVERNOSUM AGAINST HYPERCHOLESTEROLEMIAINDUCED FIBROSIS Sturny, M.1; Anguenot, L.1; Bragina, M.1; Costa-Fraga, F.1; Da Silva, R.2; Fraga-Silva, R.1; Stergiopulos, N.1 1 Ecole Polytechnique Fédérale de Lausanne, Switzerland; 2 Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte, Brazil
OPIORPHIN; A POTENTIAL KEY FACTOR IN DEVELOPMENT OF PRIAPISM ASSOCIATED WITH SICKLE CELL DISEASE Davies, K.P.1; Fu, S.2; Tar, M.T.1 1 Albert Einstein College of Medicine, United States; 2Jilin University, China
Objectives: Apelin, an endogenous circulating peptide, has been documented as an important effector on cardiovascular homeostasis, controlling vascular function and structure. Initial studies have shown that Apelin, acting through APJ receptor, also modulates penile erection. However, the role of Apelin/APJ receptor on penile structure and remodeling has never been investigated. Here we sought to evaluate the effect of Apelin treatment on hypercholesterolemic mice penile structure and its action on activated fibroblast. Material and Methods: Apolipoprotein gene-deleted (ApoE-/-) mice were fed with Western diet for 11 weeks and received Apelin-13 (2mg/kg) or vehicle during the last 3 weeks. Penile samples were obtained for histological and biochemical analyses. Furthermore, Apelin-13 actions on mouse fibroblast was also evaluated. Results: Apelin and APJ receptor were well expressed (gene and protein) within the corpus cavernosum of ApoE-/- mice, indicating local modulation of Apelin system. Interestingly, three weeks of treatment with Apelin-13 strongly reduced intracavernosal collagen
Objective(s): Priapism associated with sickle cell disease (SCD) affects several thousand men in the US and millions worldwide. We have taken the approach that SCD is a disease of hypoxia, and used both in vitro models of hypoxia and animal models to better understand the molecular events leading to priapism. By understanding the molecular events by which SCD results in priapism we hope to identify novel strategies and pharmacologic agents for its prevention or treatment. Material and Method(s): Rat corporal smooth muscle (CSM) cells in vitro were treated with CoCl2 or low oxygen tension to mimic hypoxia; changes in specific gene expression in response to hypoxia were identified by quantitative-RT-PCR and Western blotting. Using siRNA against specific genes we were able to determine the sequence of gene activation in response to hypoxia. We used BERK sickle cell mice to confirm a similar activation of genes occurred in the corpora tissue in vivo by quantitative-RT-PCR. Result(s): In vitro hypoxic conditions increased expression of genes previously associated with priapism in animal models. The rat opiorphin homologue, (called sialorphin, vcsa1;), hypoxiainducible factor-1a (Hif-1a), CD73 (50 -ribonucleotide phosphohydrolase) and A2B adenosine receptor (a2br) were increased
J Sex Med 2017;14:S1-S131
WMSM 2016 Abstracts
(Allo), hCt+allogenic-PBMCs+cyclosporineA (CsA). Additional hCt were cultured without PBMCs for 24 hours in the presence of media alone, cyclosporineA, or FK506. Tissues were evaluated by IHC and live confocal fluorescent imaging. Myography was used to examine nerve-mediated contraction and relaxation in response to electrical field stimulation. PBMC activation was assessed by flow cytometry and qPCR-array. This study was approved by the IRB. Results: Microscopy demonstrated increased caspase-3/7 activation and TUNEL staining in tissues exposed to allogeneic PBMCs, which was prevented by CsA. Flow cytometry and qPCR-array demonstrated PBMC activation when exposed to allogenic hCt, which was prevented by CsA treatment. Myography demonstrated impaired contraction and relaxation in Allo compared to Auto. CsA treated Allo had similar contraction and surprisingly impaired relaxation compared to non-treated Allo tissues. Compared to media and FK506, CsA impaired nervemediated relaxation. Conclusions: This model may be used to investigate the pathogenesis of rejection and to optimize immunosuppression for penile transplantation. Disclosure: Work supported by industry: no.
content. Accordingly, Apelin-13 enhanced total matrix metalloproteinase (MMP) activity in the mice penis, which was associated with an increased protein expression of MMP-1, MMP-3, MMP-8 and MMP-9, while tissue inhibitor of metalloproteinase(TIMP)-1, TIMP-2 and TIMP-4 were unaltered. These beneficial actions were not associated with changes in nNOS or eNOS protein expression, intracavernosal ROS content, or atherosclerotic plaque deposition. In mouse fibroblast, Apelin-13 inhibited TGFb1-induced fibroblast differentiation into myofibroblast and collagen production. Conclusion: These results suggest a local modulation of Apelin/ APJ system within the corpus cavernosum. Remarkably, Apelin13 attenuated hypercholesterolemic-induced cavernosal fibrosis by: 1) enhancing MMPs expression and activity; 2) inhibiting fibroblast differentiation into myofibroblast; and 3) diminishing collagen production. These results provide the first evidence of Apelin/APJ role on erectile tissue structure/remodeling, and point out this system as a novel potential target to alleviate intracavernosal fibrosis-related disorders. Disclosure: Work supported by industry: no.
017
018
APELIN-13 PROTECTS CORPUS CAVERNOSUM AGAINST HYPERCHOLESTEROLEMIAINDUCED FIBROSIS Sturny, M.1; Anguenot, L.1; Bragina, M.1; Costa-Fraga, F.1; Da Silva, R.2; Fraga-Silva, R.1; Stergiopulos, N.1 1 Ecole Polytechnique Fédérale de Lausanne, Switzerland; 2 Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte, Brazil
OPIORPHIN; A POTENTIAL KEY FACTOR IN DEVELOPMENT OF PRIAPISM ASSOCIATED WITH SICKLE CELL DISEASE Davies, K.P.1; Fu, S.2; Tar, M.T.1 1 Albert Einstein College of Medicine, United States; 2Jilin University, China
Objectives: Apelin, an endogenous circulating peptide, has been documented as an important effector on cardiovascular homeostasis, controlling vascular function and structure. Initial studies have shown that Apelin, acting through APJ receptor, also modulates penile erection. However, the role of Apelin/APJ receptor on penile structure and remodeling has never been investigated. Here we sought to evaluate the effect of Apelin treatment on hypercholesterolemic mice penile structure and its action on activated fibroblast. Material and Methods: Apolipoprotein gene-deleted (ApoE-/-) mice were fed with Western diet for 11 weeks and received Apelin-13 (2mg/kg) or vehicle during the last 3 weeks. Penile samples were obtained for histological and biochemical analyses. Furthermore, Apelin-13 actions on mouse fibroblast was also evaluated. Results: Apelin and APJ receptor were well expressed (gene and protein) within the corpus cavernosum of ApoE-/- mice, indicating local modulation of Apelin system. Interestingly, three weeks of treatment with Apelin-13 strongly reduced intracavernosal collagen
Objective(s): Priapism associated with sickle cell disease (SCD) affects several thousand men in the US and millions worldwide. We have taken the approach that SCD is a disease of hypoxia, and used both in vitro models of hypoxia and animal models to better understand the molecular events leading to priapism. By understanding the molecular events by which SCD results in priapism we hope to identify novel strategies and pharmacologic agents for its prevention or treatment. Material and Method(s): Rat corporal smooth muscle (CSM) cells in vitro were treated with CoCl2 or low oxygen tension to mimic hypoxia; changes in specific gene expression in response to hypoxia were identified by quantitative-RT-PCR and Western blotting. Using siRNA against specific genes we were able to determine the sequence of gene activation in response to hypoxia. We used BERK sickle cell mice to confirm a similar activation of genes occurred in the corpora tissue in vivo by quantitative-RT-PCR. Result(s): In vitro hypoxic conditions increased expression of genes previously associated with priapism in animal models. The rat opiorphin homologue, (called sialorphin, vcsa1;), hypoxiainducible factor-1a (Hif-1a), CD73 (50 -ribonucleotide phosphohydrolase) and A2B adenosine receptor (a2br) were increased
J Sex Med 2017;14:S1-S131