The Urokinase Receptor-Derived Peptide UPARANT Mitigates Angiogenesis in a Mouse Model of Laser-Induced Choroidal Neovascularization
The Urokinase Receptor-Derived Peptide UPARANT Mitigates Angiogenesis in a Mouse Model of Laser-Induced Choroidal Neovascularization
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DOI：
10.1167/iovs.15-18758
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发表时间：
2016-05-01
影响因子：
4.4
通讯作者：
Bagnoli, Paola
中科院分区：
文献类型：
--
作者：
Cammalleri, Maurizio;Dal Monte, Massimo;Bagnoli, Paola
目的.采用年龄相关性黄斑变性（AMD）小鼠模型，观察UPRANT对激光诱导的脉络膜新生血管（CNV）的抑制作用和抗炎作用。通过激光光凝诱导脉络膜新生血管形成，玻璃体内注射UPARANT。还在玻璃体内注射抗VEGF药物或全身给予UPRANT后进行了一些实验。免疫组化CD 31抗体用于评估CNV面积。定量评估伊文思蓝染料外渗。定量RT-PCR检测血视网膜外屏障标志物的转录，并用于评估血管生成和炎症标志物。蛋白质印迹法用于确定参与血管生成和炎症的基因编码转录因子的水平。尿激酶型纤溶酶原激活物（uPA）及其受体（uPAR）和甲酰肽受体（FPRs）的水平在转录和蛋白水平上进行测定。玻璃体内注射UPARANT可减少CNV面积和脉络膜渗漏。uPA/uPAR/FPR系统在CNV中上调，但不受UPARANT的影响。UPRANT恢复了激光诱导的编码血管生成和炎症标志物的转录因子的上调。因此，血管生成和炎症因子也减少。与抗VEGF药物相比，UPARANT对CNV面积的作用相似。UPRANT通过对编码血管生成和炎症基因的转录因子的作用抑制血管生成和炎症，从而减轻激光诱导的CNV。UPARANT对CNV有效的发现可能有助于将uPAR及其膜伴侣确立为治疗AMD的假定靶点。
PURPOSE. A mouse model of age-related macular degeneration (AMD) was used to investigate the anti-angiogenic and anti-inflammatory role of UPARANT in laser-induced choroidal neovascularization (CNV).METHODS. Choroidal neovascularization was induced by laser photocoagulation, and UPARANT was intravitreally injected. Some experiments were also performed after either intravitreal injection of anti-VEGF drugs or systemic administration of UPARANT. Immunohistochemistry using CD31 antibodies was used to evaluate the area of CNV. Evans blue dye extravasation was quantitatively assessed. Transcripts of markers of outer blood retinal barrier were measured by quantitative RT-PCR, also used to evaluate angiogenesis and inflammation markers. Western blot was used to determine levels of transcription factors encoding genes involved in angiogenesis and inflammation. Levels of urokinase-type plasminogen activator (uPA), its receptor (uPAR), and formyl peptide receptors (FPRs) were determined at the transcript and the protein level.RESULTS. Intravitreal UPARANT reduced the CNV area and the leakage from the choroid. The uPA/uPAR/FPR system was upregulated in CNV, but was not influenced by UPARANT. UPARANT recovered laser-induced upregulation of transcription factors encoding angiogenic and inflammatory markers. Accordingly, angiogenic and inflammatory factors were also reduced. UPARANT as compared to anti-VEGF drugs displayed similar effects on CNV area.CONCLUSIONS. UPARANT mitigates laser-induced CNV by inhibiting angiogenesis and inflammation through an action on transcription factors encoding angiogenesis and inflammatory genes. The finding that UPARANT is effective against CNV may help to establish uPAR and its membrane partners as putative targets in the treatment of AMD.