Long-term inhibition of Rho-kinase suppresses neointimal formation after stent implantation in porcine coronary arteries: involvement of multiple mechanisms.
OBJECTIVE
We recently demonstrated that Rho-kinase, an effector of the small GTPase Rho, is substantially involved in the pathogenesis of arteriosclerosis. In this study, we examined whether Rho-kinase is also involved in in-stent restenosis and if so, what mechanism is involved.


RESULTS
Pigs underwent stent implantation in the left coronary artery with or without administration of fasudil (30 mg/kg per day orally), a specific Rho-kinase inhibitor, starting 2 days before the procedure for a duration of 4 weeks. On day 28, reductions in coronary diameter and neointimal formation associated with macrophage accumulation, collagen deposition, and transforming growth factor (TGF)-beta1 expression were noted at the stent site, and all were significantly suppressed by fasudil. On day 7, fasudil significantly increased the frequency of TUNEL-positive apoptotic cells, while it tended to reduce that of bromodeoxyuridine-positive proliferating cells in the neointima. Western blot analysis on day 7 demonstrated that phosphorylations of the ezrin/radixin/moesin family (a marker of Rho-kinase activity in vivo) and protein expression of monocyte chemoattractant protein-1and bcl-2 were upregulated at the stent site and were significantly suppressed by fasudil.


CONCLUSIONS
These results indicate that long-term inhibition of Rho-kinase suppresses in-stent neointimal formation by multiple mechanisms, including reduced vascular inflammation, enhanced apoptosis, and decreased collagen deposition.
Department of Cardiovascular Medicine, Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan.
Yasuharu
Y
Matsumoto
Toyokazu
T
Uwatoku
Keiji
K
Oi
Kohtaro
K
Abe
Tsuyoshi
T
Hattori
Kunio
K
Morishige
Yasuhiro
Y
Eto
Yoshihiro
Y
Fukumoto
Kei-ichiro
K
Nakamura
Yosaburo
Y
Shibata
Takehisa
T
Matsuda
Akira
A
Takeshita
Hiroaki
H
Shimokawa
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001798:Blood Proteins / Q000378:metabolism; D002404:Catheterization / Q000009:adverse effects; D018932:Chemokine CCL2 / Q000096:biosynthesis / Q000235:genetics; D003094:Collagen / Q000378:metabolism; D023903:Coronary Restenosis / Q000201:enzymology* / Q000473:pathology / Q000517:prevention & control; D023921:Coronary Stenosis / Q000601:surgery / Q000628:therapy; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019254:Genes, bcl-2; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D016037:Single-Blind Method; D015607:Stents; D013552:Swine; D017539:Tunica Intima / Q000187:drug effects / Q000201:enzymology / Q000473:pathology*; D054460:rho-Associated Kinases; D000099149:Ezrin
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D001798:Blood Proteins; D018932:Chemokine CCL2; D003094:Collagen; D003598:Cytoskeletal Proteins; D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D017346:Protein Serine-Threonine Kinases; D019253:Proto-Oncogene Proteins c-bcl-2; D054460:rho-Associated Kinases; D000099149:Ezrin; C071058:moesin; C071776:radixin; C049347:fasudil