RhoA/Rho-kinase Suppresses Endothelial Nitric Oxide Synthase in the Penis: a Mechanism for Diabetes-associated Erectile Dysfunction

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2004 Jun 9

PMID 15184671

Citations 103

Authors

Trinity J Bivalacqua

Hunter C Champion

Mustafa F Usta

Selim Cellek

Kanchan Chitaley

R Clinton Webb

Ronald L Lewis

Thomas M Mills

Wayne J G Hellstrom

Philip J Kadowitz

Affiliations

Soon will be listed here.

Abstract

Significant impairment in endothelial-derived nitric oxide is present in the diabetic corpus cavernosum. RhoA/Rho-kinase may suppress endothelial nitric oxide synthase (eNOS). Here, we tested the hypothesis that RhoA/Rho-kinase contributes to diabetes-related erectile dysfunction and down-regulation of eNOS in the streptozotocin (STZ)-diabetic rat penis. Colocalization of Rho-kinase and eNOS protein was present in the endothelium of the corpus cavernosum. RhoA/Rho-kinase protein abundance and MYPT-1 phosphorylation at Thr-696 were elevated in the STZ-diabetic rat penis. In addition, eNOS protein expression, cavernosal constitutive NOS activity, and cGMP levels were reduced in the STZ-diabetic penis. To assess the functional role of RhoA/Rho-kinase in the penis, we evaluated the effects of an adeno-associated virus encoding the dominant-negative RhoA mutant (AAVTCMV19NRhoA) on RhoA/Rho-kinase and eNOS and erectile function in vivo in the STZ-diabetic rat. STZ-diabetic rats transfected with AAVCMVT19NRhoA had a reduction in RhoA/Rho-kinase and MYPT-1 phosphorylation at a time when cavernosal eNOS protein, constitutive NOS activity, and cGMP levels were restored to levels found in the control rats. There was a significant decrease in erectile response to cavernosal nerve stimulation in the STZ-diabetic rat. AAVT19NRhoA gene transfer improved erectile responses in the STZ-diabetic rat to values similar to control. These data demonstrate a previously undescribed mechanism for the down-regulation of penile eNOS in diabetes mediated by activation of the RhoA/Rho-kinase pathway. Importantly, these data imply that inhibition of RhoA/Rho-kinase improves eNOS protein content and activity thus restoring erectile function in diabetes.

Citing Articles

The effects of the urotensin-II receptor antagonist palosuran treatment on the corpora cavernosa of streptozotocin-induced diabetic rats.

Olukman M, Can C, Ulker S, Uyanikgil Y, Cavusoglu T, Duzenli N Asian J Urol. 2025; 12(1):116-126.

PMID: 39990066 PMC: 11840324. DOI: 10.1016/j.ajur.2024.02.010.

Idiopathic recurrent ischemic priapism: a review of current literature and an algorithmic approach to evaluation and management.

Yarak N, Khoury J, Coloby P, Bart S, Abdessater M Basic Clin Androl. 2024; 34(1):21.

PMID: 39627696 PMC: 11616154. DOI: 10.1186/s12610-024-00237-y.

Mechanisms of Xuefu Zhuyu Tang in the Treatment of Diabetic Erectile Dysfunction in Rats Through the Regulation of Vascular Endothelial Function by CaSR/PLC/PKC and MEK/ERK/RSK Pathways.

Li X, Zhang M, Feng J, Wang J, Wang K, Ju B Am J Mens Health. 2024; 18(5):15579883241277423.

PMID: 39434501 PMC: 11497541. DOI: 10.1177/15579883241277423.

RhoA/rho kinase pathway activation in age-associated endothelial cell dysfunction and thrombosis.

Palomo I, Wehinger S, Andres V, Garcia-Garcia F, Fuentes E J Cell Mol Med. 2024; 28(8):e18153.

PMID: 38568071 PMC: 10989549. DOI: 10.1111/jcmm.18153.

Hyperprolactinemia and male sexual function: focus on erectile dysfunction and sexual desire.

Corona G, Rastrelli G, Bianchi N, Sparano C, Sforza A, Vignozzi L Int J Impot Res. 2023; 36(4):324-332.

PMID: 37340146 DOI: 10.1038/s41443-023-00717-1.

References

Gong M, Gorenne I, Read P, Jia T, Nakamoto R, Somlyo A . Regulation by GDI of RhoA/Rho-kinase-induced Ca2+ sensitization of smooth muscle myosin II. Am J Physiol Cell Physiol. 2001; 281(1):C257-69. DOI: 10.1152/ajpcell.2001.281.1.C257. View

Rees R, Ralph D, Royle M, Moncada S, Cellek S . Y-27632, an inhibitor of Rho-kinase, antagonizes noradrenergic contractions in the rabbit and human penile corpus cavernosum. Br J Pharmacol. 2001; 133(4):455-8. PMC: 1572821. DOI: 10.1038/sj.bjp.0704124. View

Akingba A, Burnett A . Endothelial nitric oxide synthase protein expression, localization, and activity in the penis of the alloxan-induced diabetic rat. Mol Urol. 2002; 5(4):189-97. DOI: 10.1089/10915360152745885. View

Shimokawa H . Rho-kinase as a novel therapeutic target in treatment of cardiovascular diseases. J Cardiovasc Pharmacol. 2002; 39(3):319-27. DOI: 10.1097/00005344-200203000-00001. View

Rees R, Ziessen T, Ralph D, Kell P, Moncada S, Cellek S . Human and rabbit cavernosal smooth muscle cells express Rho-kinase. Int J Impot Res. 2002; 14(1):1-7. DOI: 10.1038/sj.ijir.3900814. View

Hurt K, Musicki B, Palese M, Crone J, Becker R, Moriarity J . Akt-dependent phosphorylation of endothelial nitric-oxide synthase mediates penile erection. Proc Natl Acad Sci U S A. 2002; 99(6):4061-6. PMC: 122648. DOI: 10.1073/pnas.052712499. View

Wang H, Eto M, Steers W, Somlyo A, Somlyo A . RhoA-mediated Ca2+ sensitization in erectile function. J Biol Chem. 2002; 277(34):30614-21. DOI: 10.1074/jbc.M204262200. View

Chitaley K, Bivalacqua T, Champion H, Usta M, Hellstrom W, Mills T . Adeno-associated viral gene transfer of dominant negative RhoA enhances erectile function in rats. Biochem Biophys Res Commun. 2002; 298(3):427-32. DOI: 10.1016/s0006-291x(02)02458-0. View

Ming X, Viswambharan H, Barandier C, Ruffieux J, Kaibuchi K, Rusconi S . Rho GTPase/Rho kinase negatively regulates endothelial nitric oxide synthase phosphorylation through the inhibition of protein kinase B/Akt in human endothelial cells. Mol Cell Biol. 2002; 22(24):8467-77. PMC: 139860. DOI: 10.1128/MCB.22.24.8467-8477.2002. View

10.

Chang S, Hypolite J, Changolkar A, Wein A, Chacko S, DiSanto M . Increased contractility of diabetic rabbit corpora smooth muscle in response to endothelin is mediated via Rho-kinase beta. Int J Impot Res. 2003; 15(1):53-62. DOI: 10.1038/sj.ijir.3900947. View

11.

Bivalacqua T, Armstrong J, Biggerstaff J, Abdel-Mageed A, Kadowitz P, Hellstrom W . Gene transfer of extracellular SOD to the penis reduces O2-* and improves erectile function in aged rats. Am J Physiol Heart Circ Physiol. 2002; 284(4):H1408-21. DOI: 10.1152/ajpheart.00770.2002. View

12.

Bivalacqua T, Usta M, Champion H, Adams D, Namara D, Abdel-Mageed A . Gene transfer of endothelial nitric oxide synthase partially restores nitric oxide synthesis and erectile function in streptozotocin diabetic rats. J Urol. 2003; 169(5):1911-7. DOI: 10.1097/01.ju.0000051881.14239.4a. View

13.

Cirino G, Sorrentino R, dEmmanuele di Villa Bianca R, Popolo A, Palmieri A, Imbimbo C . Involvement of beta 3-adrenergic receptor activation via cyclic GMP- but not NO-dependent mechanisms in human corpus cavernosum function. Proc Natl Acad Sci U S A. 2003; 100(9):5531-6. PMC: 154379. DOI: 10.1073/pnas.0931347100. View

14.

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(3):235-8. DOI: 10.1016/s0014-2999(03)01905-8. View

15.

Cellek S, Foxwell N, Moncada S . Two phases of nitrergic neuropathy in streptozotocin-induced diabetic rats. Diabetes. 2003; 52(9):2353-62. DOI: 10.2337/diabetes.52.9.2353. View

16.

Somlyo A, Somlyo A . Ca2+ sensitivity of smooth muscle and nonmuscle myosin II: modulated by G proteins, kinases, and myosin phosphatase. Physiol Rev. 2003; 83(4):1325-58. DOI: 10.1152/physrev.00023.2003. View

17.

Bivalacqua T, Usta M, Champion H, Kadowitz P, Hellstrom W . Endothelial dysfunction in erectile dysfunction: role of the endothelium in erectile physiology and disease. J Androl. 2003; 24(6 Suppl):S17-37. DOI: 10.1002/j.1939-4640.2003.tb02743.x. View

18.

Angulo J, Cuevas P, Fernandez A, Gabancho S, Allona A, Martin-Morales A . Diabetes impairs endothelium-dependent relaxation of human penile vascular tissues mediated by NO and EDHF. Biochem Biophys Res Commun. 2003; 312(4):1202-8. DOI: 10.1016/j.bbrc.2003.11.034. View

19.

Jesmin S, Sakuma I, Salah-Eldin A, Nonomura K, Hattori Y, Kitabatake A . Diminished penile expression of vascular endothelial growth factor and its receptors at the insulin-resistant stage of a type II diabetic rat model: a possible cause for erectile dysfunction in diabetes. J Mol Endocrinol. 2003; 31(3):401-18. DOI: 10.1677/jme.0.0310401. View

20.

Chacko S, Conti M, Adelstein R . Effect of phosphorylation of smooth muscle myosin on actin activation and Ca2+ regulation. Proc Natl Acad Sci U S A. 1977; 74(1):129-33. PMC: 393211. DOI: 10.1073/pnas.74.1.129. View