title: Inhibition of RhoA/Rho kinase pathway is involved in the beneficial effect of sildenafil on pulmonary hypertension.
abstract: Inhibition of the type 5 phosphodiesterase and inhibition of Rho kinase are both effective in reducing pulmonary hypertension (PH). Here we investigate whether Rho kinase inhibition is involved in the beneficial effect of the type 5 phosphodiesterase inhibitor sildenafil on PH. Chronic hypoxia-induced PH in rats is associated with an increase in RhoA activity in pulmonary artery that was maximal after 2 days (10.7+/-0.9-fold increase, n=6, P<0.001). The activity of Rho kinase assessed by measuring the level of myosin phosphatase target subunit 1 (MYPT1) phosphorylation was also increased (5.7+/-0.8-fold over control, n=8). Chronic fasudil (30 mg kg(-1) day(-1); 14 days) and sildenafil (25 mg kg(-1) day(-1); 14 days) treatments reduced PH and pulmonary cardiovascular remodelling, and inhibited the MYPT1 phosphorylation in pulmonary artery from hypoxic rats by 82.3+/-3% (n=4) and by 76.6+/-2% (n=4), respectively. The inhibitory effect of sildenafil (10 microM) on MYPT1 phosphorylation was demonstrated by the loss of actin stress fibres in vascular smooth muscle cells. However, in vitro kinase assays indicated that sildenafil had no direct inhibitory action on Rho kinase activity. Sildenafil treatment induced increased RhoA phosphorylation and association to its cytosolic inhibitory protein, guanine dissociation inhibitor (GDI) in pulmonary artery.We propose that sildenafil inhibits RhoA/Rho kinase-dependent functions in pulmonary artery through enhanced RhoA phosphorylation and cytosolic sequestration by GDI. The inhibition of intracellular events downstream of RhoA thus participates in the beneficial effect of sildenafil on PH.
authors.affiliation: INSERM U533, Université de Nantes, Institut du Thorax, Faculté des Sciences, 2 rue de la Houssiniere, BP 92208, 44322 Nantes cedex 3, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Guilluy
authors.forename: Vincent
authors.initials: V
authors.lastname: Sauzeau
authors.forename: Malvyne
authors.initials: M
authors.lastname: Rolli-Derkinderen
authors.forename: Patrice
authors.initials: P
authors.lastname: Guérin
authors.forename: Christine
authors.initials: C
authors.lastname: Sagan
authors.forename: Pierre
authors.initials: P
authors.lastname: Pacaud
authors.forename: Gervaise
authors.initials: G
authors.lastname: Loirand
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000627:therapeutic use; D000199:Actins / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002908:Chronic Disease; D003599:Cytoskeleton / Q000187:drug effects; D020730:Guanine Nucleotide Dissociation Inhibitors / Q000502:physiology; D006976:Hypertension, Pulmonary / Q000188:drug therapy* / Q000201:enzymology / Q000209:etiology; D000860:Hypoxia / Q000150:complications; D008297:Male; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology* / Q000627:therapeutic use; D054645:Protein Phosphatase 1; D011687:Purines; D051381:Rats; D017208:Rats, Wistar; D000068677:Sildenafil Citrate; D013450:Sulfones; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors*
chemical_list: D000199:Actins; D002352:Carrier Proteins; D020730:Guanine Nucleotide Dissociation Inhibitors; D010726:Phosphodiesterase Inhibitors; D010879:Piperazines; D011687:Purines; D013450:Sulfones; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000068677:Sildenafil Citrate; D010749:Phosphoprotein Phosphatases; C499850:Ppp1r12a protein, rat; D054645:Protein Phosphatase 1; D020742:rhoA GTP-Binding Protein; C049347:fasudil