Rho/rho-kinase signalling in chronically alcohol-fed mice

Rho/rho-kinase signalling in chronically alcohol-fed mice

Background/aim: The effects of chronic ethanol consumption on male sexual function are debateable, and its effects on the Rho/ Rho-kinase pathway are unknown. Therefore, we investigated smooth muscle reactivity and Rho/Rho-kinase signalling in the corpus cavernosum of ethanol-fed mice. Materials and methods: Ethanol was added to drinking water at 5% concentration in the first 2 days with 5% increment every subsequent 2 days, up to a final concentration of 20% for 45 days. Corpora cavernosa were isolated and a cumulative dose-response curve to phenylephrine was obtained. Acetylcholine (10 6 M), electrical field stimulation (EFS, 40 V, 8 16 Hz) and Y-27632 (10 6 10 5 M)-induced relaxations were compared in the control and ethanol groups. Results: Blood ethanol levels in the control and ethanol-treated mice were 1.5 ± 0.3 mg/dL and 37.4 ± 4.1 mg/dL (P < 0.001), respectively. Phenylephrine-induced contractile responses were potentiated in the ethanol group, with pD2values of 4.92 ± 0.18 and 5.71 ± 0.21 (P < 0.01) in the corpus cavernosum obtained from control and alcohol-fed mice, respectively. The relaxant responses to acetylcholine and EFS, but not Y-27632, were significantly augmented in the corpus cavernosum obtained from ethanol-treated mice. However, expression and activation of Rho-kinase remained unchanged in the alcohol group. Conclusion: Ethanol drinking may increase sensitivity to both vasoconstrictors and vasodilators in the mouse corpus cavernosum without any alteration in Rho/Rho-kinase signalling.

___

  • 1. Saito M, Broderick GA, Wein AJ, Levin RM. Effect of chronic ethanol consumption on the pharmacological response of the rabbit corpus cavernosum. Pharmacology 1994; 49: 386-391.
  • 2. Pinardi G, Brieva C, Vinet R, Penna M. Effects of chronic ethanol consumption on α-adrenergic-induced contractions in rat thoracic aorta. Gen Pharmacol 1992; 23: 245-248.
  • 3. Tirapelli CR, Al-Khoury J, Bkaily G, D’Orleans-Juste P, Lanchote VL, Uyemura SA, de Oliveira AM. Chronic ethanol consumption enhances phenylephrine-induced contraction in the isolated rat aorta. J Pharmacol Exp Ther 2005; 316: 234- 241.
  • 4. Lizarte FS, Claudino MA, Tirapelli CR, Morgueti M, Tirapelli DPC, Batalhão ME, Carnio EC, Queiroz RH, Evora PRB, Tucci S et al. Chronic ethanol consumption induces cavernosal smooth muscle dysfunction in rats. Urology 2009; 74: 1250- 1256.
  • 5. Strickland JA, Wooles WR. Effect of acute and chronic ethanol on the agonist responses of vascular smooth muscle. Eur J Pharmacol 1988; 152: 83-91.
  • 6. Utkan T, Yildiz F, Ilbay G, Ozdemirci S, Erden BF, Gacar N, Ulak G. Blood pressure and vascular reactivity to endothelin-1, phenylephrine, serotonin and acetylcholine following chronic alcohol consumption in vitro. Fundam Clin Pharm 2001; 15: 157-165.
  • 7. Kim HJ, Sohng I, Lee G, Kim JJ, Koh SK. Effects of acetaldehyde on responses of rabbit corpus cavernosal smooth muscle. J Korean Med Sci 2000; 15: 295-298.
  • 8. Buyukafsar K, Un I. Effects of the Rho-kinase inhibitors, Y-27632 and fasudil, on the corpus cavernosum from diabetic mice. Eur J Pharmacol 2003; 472: 235-238.
  • 9. Chitaley K, Wingard C J, Webb RC, Branam H, Stopper VS, Lewis RW, Mills TM. Antagonism of Rho-kinase stimulates rat penile erection via a nitric oxide-independent pathway. Nat Med 2001; 7: 119-122.
  • 10. Buyukafsar K, Levent A, Ark M. Expression of Rho-kinase and its functional role in the contractile activity of the mouse vas deferens. Br J Pharmacol 2003; 140: 743-749.
  • 11. Levent A, Buyukafsar K. Expression of Rho-kinase (ROCK-1 and ROCK-2) and its substantial role in the contractile activity of the sheep ureter. Br J Pharmacol 2004; 143: 431-437.
  • 12. Turna B, Cinar MG, Canda AE, Orhan EC, Tiftik NR, Nazli O, Buyukafsar K. Role of Rho-kinase in contractions of ureters from rabbits with unilateral ureteric obstruction. BJU Int 2007; 100: 1166-1171.
  • 13. Bivalacqua TJ, Champion HC, Usta MF, Cellek S, Chitaley K, Webb RC, Lewis RL, Mills TM, Hellstrom WJ, Kadowitz PJ. RhoA/Rho-kinase suppresses endothelial nitric oxide synthase in the penis: a mechanism for diabetes-associated erectile dysfunction. Proc Natl Acad Sci USA 2004; 101: 9121-9126.
  • 14. Altura BM, Edgarian H, Altura BT. Differential effects of ethanol and mannitol on contraction of arterial smooth muscle. J Pharmacol Exp Ther 1976; 197: 352-361.
  • 15. Tirapelli CR, Leone AF, Coelho EB, Resstel LB, Corrêa FM, Lanchote VL, Uyemura SA, Padovan CM, de Oliveira AM. Ethanol consumption increases blood pressure and alters the responsiveness of the mesenteric vasculature in rats. J Pharm Pharmacol 2008; 60: 331-341.
  • 16. Werber AH, Morgan RA, Zhou P, Yang C. Intracellular mechanisms of constriction of rat aorta by ethanol. Alcohol 1997; 14: 351-360.
  • 17. Buyukafsar K, Gocmen C, Secilmis A, Karatas Y, Gokturk S, Kalyoncu NI. Evidence that the nitrergic neurotransmitter and endothelium-derived relaxing factor might be S-nitrosothiols in the mouse corpus cavernosum. Acta Med Okayama 1999; 53: 209-215.
  • 18. Davda RK, Chandler LJ, Crews FT, Guzman NJ. Ethanol enhances the endothelial nitric oxide synthase response to agonists. Hypertension 1993; 21: 939-943.
  • 19. Greenberg SS, Xie J, Wang Y, Kolls J, Shellito J, Nelson S, Summer WS. Ethanol relaxes pulmonary artery by release of prostaglandin and nitric oxide. Alcohol 1993; 10: 21-29.
  • 20. Venkov CD, Myers PR, Tanner MA, Su M, Vaughan DE. Ethanol increases endothelial nitric oxide production through modulation of nitric oxide synthase expression. Thromb Haemostasis 1999; 81: 638-642.
  • 21. Kuhlmann CR, Li F, Lüdders DW, Schaefer CA, Most AK, Backenköhler U, Neumann T, Tillmanns H, Waldecker B, Erdogan A et al. Dose-dependent activation of Ca2+-activated K+ channels by ethanol contributes to improved endothelial cell functions. Alcohol Clin Exp Res 2004; 28: 1005-1011.
  • 22. Hendrickson RJ, Cahill PA, Sitzmann JV, Redmond EM. Ethanol enhances basal and flow-stimulated nitric oxide synthase activity in vitro by activating an inhibitory guanine nucleotide binding protein. J Pharmacol Exp Ther 1999; 289: 1293-1300.
  • 23. Zelickson BR, Benavides GA, Johnson MS, Chacko BK, Venkatraman A, Landar A, Betancourt AM, Bailey SM, Darley-Usmar VM. Chronic alcohol consumption results in inducible nitric oxide synthase (iNOS) and decreased activities of mitochondrial respiratory enzymes. Biochim Biophys Acta 2011; 1807: 1573-1582.
  • 24. Baraona E, Zeballos GA, Shoichet L, Mak KM, Lieber CS. Ethanol consumption increases nitric oxide production in rats, and its peroxynitrite-mediated toxicity is attenuated by polyenylphosphatidylcholine. Alcohol Clin Exp Res 2002; 26: 883-889.
  • 25. McKim SE, Gäbele E, Isayama F, Lambert JC, Tucker LM, Wheeler MD, Connor HD, Mason RP, Doll MA, Hein DW et al. Inducible nitric oxide synthase is required in alcohol-induced liver injury: studies with knockout mice. Gastroenterology 2003; 125: 1834-1844.
  • 26. Abeysinghe HR, Clancy J, Qiu Y. Comparison of endothelin1-mediated tissue tension and calcium mobilization effects in isolated rabbit corpus cavernosum. Urology 2002; 60: 925-930.
  • 27. Berridge MJ. Smooth muscle cell calcium  activation mechanisms. J Physiol 2008; 586: 5047-5061.
  • 28. Lopez-Jaramillo P, Gonzalez MC, Palmer RM, Moncada S. The crucial role of physiological Ca2+ concentrations in the production of endothelial nitric oxide and the control of vascular tone. Br J Pharmacol 1990; 101: 489-493.
  • 29. Wickramasinghe SN, Hasan R, Pearson JD. A possible acetaldehyde-mediated cardioprotective mechanism. Alcohol Alcohol 1996; 31: 309-311.
  • 30. Ertug PU, Buyukafsar K, Kumcu EK, Göçmen C, Seçilmis A, Singirik E, Dikmen A, Baysal F. Effects of some divalent cations on nitrergic relaxations in the mouse corpus cavernosum. Fundam Clin Pharm 2001; 15: 343-348.
  • 31. Okamura T, Fujioka H, Ayajiki K. Effects of calcium antagonists on the nitrergic nerve function in canine corpus cavernosum. Jpn J Pharmacol 2001; 87: 208-213.