The molecular mechanism of actinomycin D in preventing neointimal formation in rat carotid arteries after balloon injury.
The pathological mechanism of restenosis is primarily attributed to excessive proliferation of vascular smooth muscle cells (SMC). Actinomycin D has been regarded as a potential candidate to prevent balloon injury-induced neointimal formation. To explore its molecular mechanism in regulating cell proliferation, we first showed that actinomycin D markedly reduced the SMC proliferation via the inhibition of BrdU incorporation at 80 nM. This was further supported by the G1-phase arrest using a flowcytometric analysis. Actinomycin D was extremely potent with an inhibitory concentration IC50 at 0.4 nM, whereas the lethal dose LD50 was at 260 microM. In an in vivo study, the pluronic gel containing 80 nM and 80 microM actinomycin D was applied topically to surround the rat carotid adventitia; the thickness of neointima was substantially reduced (45 and 55%, respectively). The protein expression levels of proliferating cell nuclear antigen (PCNA), focal adhesion kinase (FAK), and Raf were all suppressed by actinomycin D. Extracellular signal-regulated kinases (Erk) involved in cell-cycle arrest were found to increase by actinomycin D. These observations provide a detailed mechanism of actinomycin D in preventing cell proliferation thus as a potential intervention for restenosis.
Wu
C H
CH
Department of Pharmacology, School of Medicine, China Medical University, 91 Hsueh-Shieh Road, Taichung, 404, Taiwan. chhswu@mail.cmu.edu.tw
Pan
J S
JS
Chang
W C
WC
Hung
J S
JS
Mao
Simon J T
SJ
D000818:Animals; D002339:Carotid Arteries / Q000187:drug effects / Q000378:metabolism*; D002404:Catheterization / Q000009:adverse effects*; D002453:Cell Cycle; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003609:Dactinomycin / Q000494:pharmacology*; D007928:Lethal Dose 50; D009131:Muscle, Smooth, Vascular / Q000187:drug effects; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D017539:Tunica Intima / Q000187:drug effects* / Q000473:pathology
D018809:Proliferating Cell Nuclear Antigen; D003609:Dactinomycin