Dong Zhenyu, Noda Kousuke, Kanda Atsuhiro, Fukuhara Junichi, Ando Ryo, Murata Miyuki, Saito Wataru, Hagiwara Masatoshi, Ishida Susumu
Department of Ophthalmology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Mol Vis. 2013;19:536-43. Epub 2013 Mar 5.
To investigate the applicability of serine/arginine-rich protein kinase (SRPK)-specific inhibitor, SRPIN340, for attenuation of choroidal neovascularization (CNV) formation using a mouse model.
Laser photocoagulation was performed to induce CNV in C57BL/6J mice, followed by intravitreal injection of SRPIN340 or vehicle. Seven days after the treatment, the CNV size was evaluated using a flatmount technique. Protein levels of vascular endothelial growth factor (VEGF) and inflammation-associated molecules, such as monocyte chemoattractant protein (MCP)-1 and intercellular adhesion molecule (ICAM)-1, in the retinal pigment epithelium-choroid complex were measured with enzyme-linked immunosorbent assay. Expression levels of total Vegf, exon 8a-containing Vegf isoforms, and F4/80 (a specific marker for macrophage) were assessed using real-time PCR.
SRPIN340 inhibited CNV formation in a dose-dependent manner. Compared with the vehicle, SRPIN340 significantly decreased the protein levels of VEGF, MCP-1, ICAM-1, and consequently inhibited macrophage infiltration. Furthermore, SRPIN340 suppressed the gene expression levels of total Vegf and exon 8a-containing Vegf isoforms.
SRPIN340, a specific inhibitor of SRPK, suppressed Vegf expression and attenuated CNV formation. Our data suggest the possibility that SRPIN340 is applicable for neovascular age-related macular degeneration as a novel chemical therapeutics.
利用小鼠模型研究富含丝氨酸/精氨酸蛋白激酶（SRPK）特异性抑制剂SRPIN340在减轻脉络膜新生血管（CNV）形成方面的适用性。
对C57BL/6J小鼠进行激光光凝诱导CNV形成，随后玻璃体内注射SRPIN340或赋形剂。治疗7天后，采用平铺技术评估CNV大小。用酶联免疫吸附测定法测量视网膜色素上皮-脉络膜复合体中血管内皮生长因子（VEGF）以及炎症相关分子如单核细胞趋化蛋白（MCP）-1和细胞间黏附分子（ICAM）-1的蛋白水平。使用实时PCR评估Vegf总量、含外显子8a的Vegf异构体以及F4/80（巨噬细胞特异性标志物）的表达水平。
SRPIN340以剂量依赖性方式抑制CNV形成。与赋形剂相比，SRPIN340显著降低了VEGF、MCP-1、ICAM-1的蛋白水平，从而抑制了巨噬细胞浸润。此外，SRPIN340抑制了Vegf总量和含外显子8a的Vegf异构体的基因表达水平。
SRPK的特异性抑制剂SRPIN340抑制了Vegf表达并减轻了CNV形成。我们的数据表明SRPIN340作为一种新型化学治疗药物适用于新生血管性年龄相关性黄斑变性的可能性。
