Shimokawa H, Morishige K, Miyata K, Kandabashi T, Eto Y, Ikegaki I, Asano T, Kaibuchi K, Takeshita A
Department of Cardiovascular Medicine, Kyushu University Graduate School of Medical Sciences, 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Cardiovasc Res. 2001 Jul;51(1):169-77. doi: 10.1016/s0008-6363(01)00291-7.
We recently demonstrated that Rho-kinase/ROK/ROCK is functionally upregulated at the arteriosclerotic coronary lesions and plays a key role for coronary vasospastic responses in our porcine model with interleukin (IL)-1beta. In the present study, we tested our hypothesis that Rho-kinase is involved in the pathogenesis of coronary arteriosclerosis per se in our porcine model.
Segments of the left porcine coronary artery were chronically treated from the adventitia with IL-1beta. Two weeks after the procedure, coronary stenotic lesions with constrictive remodeling and vasospastic response to serotonin were noted at the IL-1beta-treated site, as previously reported. Then, animals were randomly divided into two groups; one group was treated with fasudil for 8 weeks followed by 1 or 4 weeks of washout period and another group served as a control. After oral absorption, fasudil is metabolized to hydroxyfasudil that is a specific inhibitor of Rho-kinase.
In the fasudil group, coronary stenosis and vasospastic response were progressively reduced in vivo, while the coronary hyperreactivity was abolished both in vivo and in vitro. Furthermore, Western blot analysis showed that in the fasudil group, the Rho-kinase activity (as evaluated by the extent of phosphorylation of myosin binding subunit of myosin phosphatase, one of the major substrates of Rho-kinase) was significantly reduced, while histological examination demonstrated a marked regression of the coronary constrictive remodeling.
These results indicate that Rho-kinase is substantially involved in constrictive remodeling and vasospastic activity of the arteriosclerotic coronary artery, both of which could be reversed by long-term inhibition of the molecule in vivo. Thus, Rho-kinase may be regarded as a novel therapeutic target for arteriosclerotic vascular disease.
我们最近证明，在动脉粥样硬化性冠状动脉病变中，Rho激酶/ROK/ROCK功能上调，并且在我们的白细胞介素（IL）-1β猪模型中，其在冠状动脉痉挛反应中起关键作用。在本研究中，我们检验了以下假设：在我们的猪模型中，Rho激酶参与冠状动脉粥样硬化本身的发病机制。
从外膜对猪左冠状动脉节段进行IL-1β长期处理。如先前报道，处理两周后，在IL-1β处理部位观察到具有收缩性重塑和对5-羟色胺的血管痉挛反应的冠状动脉狭窄病变。然后，将动物随机分为两组；一组用法舒地尔治疗8周，随后有1或4周的洗脱期，另一组作为对照。口服吸收后，法舒地尔代谢为羟基法舒地尔，后者是Rho激酶特异性抑制剂。
在法舒地尔组中，冠状动脉狭窄和血管痉挛反应在体内逐渐减轻，而冠状动脉高反应性在体内和体外均被消除。此外，蛋白质印迹分析显示，在法舒地尔组中，Rho激酶活性（通过肌球蛋白磷酸酶的肌球蛋白结合亚基的磷酸化程度评估，肌球蛋白磷酸酶是Rho激酶的主要底物之一）显著降低，而组织学检查显示冠状动脉收缩性重塑明显消退。
这些结果表明，Rho激酶在动脉粥样硬化性冠状动脉的收缩性重塑和血管痉挛活动中起重要作用，体内长期抑制该分子可逆转这两种情况。因此，Rho激酶可被视为动脉粥样硬化性血管疾病的新治疗靶点。
