PURPOSE: To investigate the tissue distribution of PDE5 isoforms in type 2 diabetic rat penile tissues. MATERIALS AND METHODS: We prepared ten male Otsuka Long-Evans Tokushima Fatty (OLETF) rats, which develop NIDDM naturally, and ten control male Long-Evans Tokushima Fatty (LETO) rats. An oral glucose tolerance test confirmed diabetes development in OLETF rats at 26 weeks. At 42 weeks, we checked serum glucose, testosterone, triglyceride, insulin, and adiponectin before sacrifice. We performed semi-quantitative multiplex RT-PCR for rat PDE5, PDE5A1, and PDE5A2. Immunohistochemistry was performed using mouse monoclonal anti-cGB-PDE5 and anti-smooth muscle alpha-actin. RESULTS: OLETF rats were significantly more hyperglycemic, hypogonadal, hyperinsulinemic, hypercholesterolemic, hypertriglycemic, and had lower adiponectin levels than LETO rats. Levels of PDE5 mRNA were decreased in OLETF rats, but there were no changes in PDE5A1 or PDE5A2 mRNA levels. CONCLUSION: Diabetes may contribute to decreased expression of PDE5 mRNA, but not PDE5A1 or PDE5A2, in rat penile tissue. Furthermore, serum free testosterone was decreased in diabetic rats. PDE5 has an important role in the development of diabetic erectile dysfunction, but it is not clear whether PDE5A1 and PDE5A2 gene have specific roles.
Actins ; Adiponectin ; Animals ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; Diabetes Mellitus ; Diabetes Mellitus, Type 2 ; Erectile Dysfunction ; Glucose ; Glucose Tolerance Test ; Humans ; Immunohistochemistry ; Insulin ; Male ; Mice ; Muscles ; Penis ; Protein Isoforms ; Rats ; Rats, Inbred OLETF ; RNA, Messenger ; Testosterone ; Tissue Distribution

PURPOSE: To investigate the tissue distribution of PDE5 isoforms in type 2 diabetic rat penile tissues. MATERIALS AND METHODS: We prepared ten male Otsuka Long-Evans Tokushima Fatty (OLETF) rats, which develop NIDDM naturally, and ten control male Long-Evans Tokushima Fatty (LETO) rats. An oral glucose tolerance test confirmed diabetes development in OLETF rats at 26 weeks. At 42 weeks, we checked serum glucose, testosterone, triglyceride, insulin, and adiponectin before sacrifice. We performed semi-quantitative multiplex RT-PCR for rat PDE5, PDE5A1, and PDE5A2. Immunohistochemistry was performed using mouse monoclonal anti-cGB-PDE5 and anti-smooth muscle alpha-actin. RESULTS: OLETF rats were significantly more hyperglycemic, hypogonadal, hyperinsulinemic, hypercholesterolemic, hypertriglycemic, and had lower adiponectin levels than LETO rats. Levels of PDE5 mRNA were decreased in OLETF rats, but there were no changes in PDE5A1 or PDE5A2 mRNA levels. CONCLUSION: Diabetes may contribute to decreased expression of PDE5 mRNA, but not PDE5A1 or PDE5A2, in rat penile tissue. Furthermore, serum free testosterone was decreased in diabetic rats. PDE5 has an important role in the development of diabetic erectile dysfunction, but it is not clear whether PDE5A1 and PDE5A2 gene have specific roles.
Actins ; Adiponectin ; Animals ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; Diabetes Mellitus ; Diabetes Mellitus, Type 2 ; Erectile Dysfunction ; Glucose ; Glucose Tolerance Test ; Humans ; Immunohistochemistry ; Insulin ; Male ; Mice ; Muscles ; Penis ; Protein Isoforms ; Rats ; Rats, Inbred OLETF ; RNA, Messenger ; Testosterone ; Tissue Distribution

Cells, Cultured ; Cyclic Nucleotide Phosphodiesterases, Type 2 ; antagonists & inhibitors ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Glucosides ; pharmacology ; Humans ; Lung ; cytology ; embryology ; Phenols ; pharmacology ; Phosphodiesterase Inhibitors ; pharmacology ; Tumor Suppressor Protein p53 ; metabolism

The purpose of this study was to identify the effect of sildenafil citrate on IL-1 beta induced nitric oxide (NO) synthesis and iNOS expression in human synovial sarcoma SW982 cells. IL-1 beta stimulated the cells to generate NO in both dose- and time-dependent manners. The IL-1 beta -induced NO synthesis was inhibited by guanylate cyclase (GC) inhibitor, LY83583. When the cells were treated with 8-bromo-cGMP, a hydrolyzable analog of cGMP, NO synthesis was increased upto 5-fold without IL-1 beta treatment suggesting that cGMP is an essential component for increasing the NO synthesis. Synoviocytes and chondrocytes contain strong cGMP phosphodiesterase (PDE) activity, which has biochemical features of PDE5. When SW982 cells were pretreated with sildenafil citrate (Viagra), a PDE5 specific inhibitor, sildenafil citrate significantly inhibited IL-1 beta -induced NO synthesis and iNOS expressions. From this result, we noticed that PDE5 activity is required for IL-1 beta -induced NO synthesis and iNOS expressions in human synovial sarcoma cells, and sildenafil citrate may be able to suppress an inflammatory reaction of synovium through inhibition of NO synthesis and iNOS expression by cytokines.
Anti-Inflammatory Agents/immunology/pharmacology ; Cell Line, Tumor ; Cyclic GMP/analogs & derivatives/immunology/metabolism ; Cyclic Nucleotide Phosphodiesterases, Type 2/antagonists & inhibitors/metabolism ; Humans ; Interleukin-1beta/*metabolism ; Male ; Nitric Oxide/*biosynthesis/genetics/immunology ; Nitric Oxide Synthase Type II/*biosynthesis/genetics/immunology ; Phosphodiesterase Inhibitors/immunology/*pharmacology ; Piperazines/immunology/*pharmacology ; Purines/immunology/pharmacology ; Signal Transduction/drug effects/genetics/immunology ; Sulfones/immunology/*pharmacology ; Synovial Membrane/enzymology/immunology