Tat PTD-Endostatin-RGD: A novel protein with anti-angiogenesis effect in retina via eye drops
Tat PTD-Endostatin-RGD: A novel protein with anti-angiogenesis effect in retina via eye drops
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Tat PTD-Endostatin-RGD：一种通过滴眼剂在视网膜中具有抗血管生成作用的新型蛋白质
DOI：
10.1016/j.bbagen.2016.05.031
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发表时间：
2016-10-01
影响因子：
3
通讯作者：
Wang, Fengshan
中科院分区：
文献类型：
--
作者：
Li, Yan;Li, Lian;Wang, Fengshan
背景：糖尿病视网膜病变是导致失明的主要原因。目的是设计一种新的融合蛋白Tat PTD-Endostatin-RGD，通过滴眼液治疗视网膜新生血管，而不是传统的玻璃体内注射治疗方法。方法：采用鸡胚绒毛尿囊膜实验、伤口愈合实验和试管形成实验，评价其体外抗血管生成能力。体外构建角膜屏障和血视网膜屏障，观察Tat - PTD-Endostatin-RGD的渗透能力。Western blot法检测s -亚硝基-n -乙酰青霉胺刺激大鼠视网膜微血管内皮细胞中整合素α (v) β(3)的表达水平。通过评价s -亚硝基-n -乙酰青霉胺对血视网膜屏障的穿透能力，研究Tat PTD-Endostatin-RGD与整合素α的结合亲和力。在体内氧致视网膜病变模型中进一步进行药效学和疗效分析。通过滴眼液对C57BL/6小鼠模型进行药代动力学研究。结果：ptd - endostin - rgd具有较强的抗血管生成活性和穿透这两种屏障的能力。Western blot结果显示，s -亚硝基-n -乙酰青霉胺以剂量依赖的方式上调整合素α (v) β(3)的表达水平。结果表明，PTD-Endostatin-RGD对s -亚硝基-n -乙酰青霉胺处理的大鼠视网膜微血管内皮细胞具有较高的亲和力。结果表明ptd -内皮抑素- rgd可通过滴眼液抑制视网膜异常血管生成。结论：PTD-Endostatin-RGD具有高穿透眼屏障的能力，与整合素α (v) β(3)特异性结合，能有效抑制异常血管生成。一般意义：ptd -内皮抑素rgd是一种通过滴眼液治疗眼底新生血管疾病的有效新药。(C) 2016 Elsevier B.V.版权所有
Background: Diabetic retinopathy is a leading cause of blindness. The objective was to design a novel fusion protein, Tat PTD-Endostatin-RGD, to treat retinal neovascularization via eye drops instead of traditional intravitreal injection trepapeutical methods.Method: The anti-angiogenesis ability was evaluated in vitro by chick embryo chorioallantoic membrane assay, wound healing assay and tube formation assay. Corneal barrier and blood-retina barrier were constructed in vitro to investigate the penetration ability of Tat PTD-Endostatin-RGD. Western blot was used to detect the integrin alpha(v)beta(3), expression level in rat retina microvascular endothelial cells which was stimulated by S-nitroso-N-acetylpenicillamine. The binding affinity of Tat PTD-Endostatin-RGD to integrin alpha was investigated by evaluating the penetration ability on blood-retina barriers treated with S-nitroso-N-acetylpenicillamine. The pharmacodynamics and efficacy analysis were further carried out in the oxygen-induced retinopathy model in vivo. In addition, the pharmacokinetic profile via eye drops was studied on a C57BL/6 mice model.Result: Tat PTD-Endostatin-RGD showed high anti-angiogenesis activity and high ability to penetrate these two barriers in vitro. The Western blot results indicated S-nitroso-N-acetylpenicillamine upregulated the expression level of integrin alpha(v)beta(3) in a dose-dependent manner. Tat PTD-Endostatin-RGD showed a high affinity to rat retina microvascular endothelial cells treated with S-nitroso-N-acetylpenicillamine. The results showed that Tat PTD-Endostatin-RGD could inhibit abnormal angiogenesis in retina via eye drops.Conclusion: Tat PTD-Endostatin-RGD showed high penetration ability through ocular barriers, bound specifically to integrin alpha(v)beta(3) and effectively inhibited the abnormal angiogenesis.General significance: Tat PTD-Endostatin-RGD represents a potent novel drug applied via eye drops for fundus oculi neovascularization diseases. (C) 2016 Elsevier B.V. All rights reserved.