Effects of rho-associated protein kinase inhibitor Y-27632 on intraocular pressure and outflow facility.
Effects of rho-associated protein kinase inhibitor Y-27632 on intraocular pressure and outflow facility.
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发表时间：
2001
影响因子：
4.4
通讯作者：
M. Honjo;H. Tanihara;M. Inatani;N. Kido;T. Sawamura;B. Yue;S. Narumiya;Yoshihito Honda
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文献类型：
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作者：
M. Honjo;H. Tanihara;M. Inatani;N. Kido;T. Sawamura;B. Yue;S. Narumiya;Yoshihito Honda
目的阐明Rho相关蛋白激酶（ROCK）在调节兔眼内压（IOP）和流出道功能中的作用。方法采用ROCK特异性抑制剂Y-27632。在兔中局部、前房内或玻璃体内给予Y-27632之前和之后，测定IOP、流出设施和瞳孔直径。蛋白质印迹分析用于鉴定人小梁网（TM）细胞和牛睫状肌（CM）组织中的特异性ROCK亚型。应用细胞生物学技术研究了TM细胞的细胞形态及肌动蛋白丝和黏着斑蛋白的分布。在灌注室中测量Y-27632给药后卡巴胆碱（Cch）诱导的离体牛CM条的收缩。结果：Y-27632能显著降低兔眼眼压，并呈剂量依赖性。在Y-27632给药眼中还观察到流出道灵活性增加和瞳孔大小扩张。Western blot分析显示p160 ROCK在人TM细胞和牛CM组织中的存在。在培养的人TM细胞中，暴露于Y-27632引起细胞体的收缩和变圆以及肌动蛋白束的破坏和粘着斑形成的损害。此外，Y-27632还可抑制Cch诱导的离体牛CM条收缩。结论：Y-27632给药引起IOP降低和流出道功能增加。体外实验表明，Y-27632的降眼压作用可能与TM细胞的细胞行为改变和CM收缩的松弛有关。这些研究表明，ROCK抑制剂可能有很大的潜力被开发用于治疗青光眼和其他眼部疾病。
PURPOSE To elucidate the roles of Rho-associated protein kinase (ROCK) in regulating intraocular pressure (IOP) and outflow facility in the rabbit eye. METHODS A specific ROCK inhibitor Y-27632 was used. The IOP, the outflow facility, and the pupil diameter were determined before and after the topical, intracameral, or intravitreal administration of Y-27632 in rabbits. Western blot analysis was used to identify specific ROCK isoform in human trabecular meshwork (TM) cells and bovine ciliary muscle (CM) tissues. The cell morphology and distribution of actin filaments and vinculin in TM cells were studied by cell biology techniques. Carbachol (Cch)-induced contraction of isolated bovine CM strips after administration of Y-27632 was measured in a perfusion chamber. RESULTS In rabbit eyes, administration of Y-27632 resulted in a significant decrease in IOP in a dose-dependent manner. An increase of the outflow facility and pupil size dilation was also observed in Y-27632-treated eyes. Western blot analysis revealed the presence of p160ROCK in human TM cells and bovine CM tissues. In cultured human TM cells, exposure to Y-27632 caused retraction and rounding of cell bodies as well as disruption of actin bundles and impairment of focal adhesion formation. Y-27632 in addition inhibited Cch-induced contraction of isolated bovine CM strips. CONCLUSIONS Administration of Y-27632 caused a reduction in IOP and an increase in the outflow facility. The in vitro experiments suggest that the IOP-lowering effects of Y-27632 may be related to the altered cellular behavior of TM cells and relaxation of CM contraction. These studies suggest that ROCK inhibitors may have great potential to be developed for treatment of glaucoma and other ocular diseases.