Inhibition of cGMP phosphodiesterase 5 suppresses matrix metalloproteinase-2 production in pulmonary artery smooth muscles cells.
1. It has been shown that the beneficial effects of phosphodiesterase (PDE) 5 inhibition on pulmonary hypertension (PH) are associated with the induction of vascular relaxation and suppression of the proliferation of pulmonary artery smooth muscle cells (PASMC). In the present study, we investigated whether PDE5 inhibition affects the production and/or secretion of matrix metalloproteinases (MMPs) in PASMC, resulting in extracellular matrix remodelling in the pulmonary vasculature and, thus, the development of PH. 2. Primary cultured PASMC were stimulated with endothelin (ET)-1 and MMP-2 production and RhoA activation were then determinded using gelatin zymography and a GTP-bound RhoA assay, respectively. The effects of the selective PDE5 inhibitor sildenafil and subsequent protein kinase G-specific inhibitor Rp-8Br-cGMPs on MMP-2 production and RhoA activation were further exmamined. 3. Endothelin-1 (1-1000 nmol/L) concentration-dependently stimulated MMP-2 production and/or secretion in primary cultured PASMC, with 100 nmol/L ET-1 causing a 2.41-fold increase in MMP-2 production compared with control (P < 0.01). This increase in MMP-2 production was accompanied by RhoA activation, which was abolished by preincubation of cells with 10 micromol/L Y27632, an inhibitor of Rho-associated kinase (ROCK). Furthermore, 10 micromol/L Y27632 abolished the ET-1-induced production of MMP-2. 4. The selective PDE5 inhibitor sildenafil (0.1-1 micromol/L) concentration-dependently reduced the increased MMP-2 production induced by 100 nmol/L ET-1. Specifically, in the presence of 1 micromol/L sildenafil, the 100 nmol/L ET-1-induced increase in MMP-2 production was only increased 1.3-fold over that of the control (P < 0.01 vs 100 nmol/L ET-1-stimulated cells). 5. Suppression of RhoA activation was found to mediate the inhibitory effect of sildenafil on ET-1-induced increases in MMP-2 production. Furthermore, the protein kinase G-specific inhibitor Rp-8Br-cGMPs reversed the inhibitory effects of sildenafil on RhoA activation and MMP-2 production. 6. The results of the present study indicate that PDE5 inhibition suppresses RhoA/ROCK-mediated MMP-2 production by PASMC, which may contribute to the regulation of pulmonary vascular remodelling. Thus, PDE5 inhibition may benefit patients with PH through multiple mechanisms of action.
Sun
Xiu-Zhen
XZ
Department of Respiratory Medicine, The Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Li
Zong-Fang
ZF
Liu
Yun
Y
Fang
Ping
P
Li
Man-Xiang
MX
D000818:Animals; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000037:antagonists & inhibitors* / Q000378:metabolism; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5 / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis; D061965:Matrix Metalloproteinase Inhibitors; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000201:enzymology*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000201:enzymology*; D058986:Phosphodiesterase 5 Inhibitors / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D061965:Matrix Metalloproteinase Inhibitors; D058986:Phosphodiesterase 5 Inhibitors; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; D020778:Matrix Metalloproteinase 2; D006152:Cyclic GMP