Metformin In Vitro and In Vivo Increases Adenosine Signaling in Rabbit Corpora Cavernosa

IntroductionIn subjects with erectile dysfunction responding poorly to sildenafil, metformin was reported to improve erections.AimsThe aim of this study is to investigate metformin's mechanism of action on erectile function, particularly focusing on adenosine (ADO) and nitric oxide (NO) signaling in an animal model of high‐fat diet (HFD)‐induced metabolic syndrome.MethodsIn vitro contractility studies of penile strips. Penile expression of genes related to ADO or NO signaling was also evaluated.Main Outcome MeasureIn vitro contractility studies were used to investigate the effect of in vivo and ex vivo metformin administration on ADO‐ or acetylcholine (Ach)‐induced relaxation of penile strips from HFD as compared with animals fed a regular diet (RD).ResultsExpression of ADO receptor type 3 (A3R), ADO deaminase (ADA), AMP deaminase type 1 (AMPD1), and 2 (AMPD2) was decreased in HFD as compared with RD. Accordingly, in HFD the ADO relaxant effect was potentiated as compared with RD (P < 0.02). In vivo metformin treatment in both RD and HFD significantly increased the ADO relaxing effect (P < 0.0001 and P < 0.01, respectively, vs. relative untreated groups) although to a different extent. In fact, the half‐maximal inhibitory concentration (IC50)/IC50 ratio in RD increased fourfold vs. HFD (RD IC50 ratio = 13.75 ± 2.96; HFD IC50 ratio = 2.85 ± 0.52). In corpora cavernosa (CC) from HFD, in vivo metformin (i) normalized A3R, ADA, and AMPD1; (ii) further decreased AMPD2; (iii) increased dimethylarginine dimethylamino‐hydrolase; and (iv) partially restored impaired Ach‐induced relaxation. Ex vivo metformin time and dose dependently increased the relaxant effect of ADO in RD. The potentiating effect of metformin on ADO‐induced relaxation was significantly reduced by preincubation with NO synthase inhibitor Nω‐Nitro‐L‐arginine methyl ester hydrochloride (L‐NAME). Interestingly, in vivo testosterone supplementation in HFD rabbits (i) increased penile expression of endothelial NO synthase and AMPD2 and (ii) restored metformin's potentiating effect on ADO‐induced relaxation up to RD level.ConclusionMetformin in vivo and ex vivo increases ADO signaling in CC, most probably interfering with NO formation and ADO breakdown. Vignozzi L, Filippi S, Comeglio P, Cellai I, Morelli A, Rastrelli G, Maneschi E, Mannucci E, and Maggi M. Metformin in vitro and in vivo increases adenosine signaling in rabbit corpora cavernosa. J Sex Med 2014;11:1694–1708.