VEGF receptor blockade markedly reduces retinal microglia/macrophage infiltration into laser-induced CNV.
VEGF receptor blockade markedly reduces retinal microglia/macrophage infiltration into laser-induced CNV.
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DOI：
10.1371/journal.pone.0071808
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Vinores SA
中科院分区：
文献类型：
--
作者：
Huang H;Parlier R;Shen JK;Lutty GA;Vinores SA
尽管阻断血管内皮生长因子在湿性老年性黄斑变性(AMD)中有积极作用，但阻断其受体的作用尚不清楚。本实验观察了血管内皮生长因子受体(VEGFR)1和/或2受体拮抗剂对激光诱导的湿性AMD脉络膜新生血管(CNV)小胶质/巨噬细胞浸润的影响。激光后3d、7d、14d用激光捕获显微切割法分离CNV病变，分别用RT-PCR和免疫荧光染色检测CNV的mRNA和蛋白表达。用VEGFR1或R2的中和抗体和小胶质细胞抑制剂米诺环素腹腔注射(Ip)。用抗CD11b、CD45和Iba1抗体确认视网膜小胶质细胞/巨噬细胞在RPE/脉络膜平台或视网膜横切面的细胞特性。计数CD11b(+)、CD45(+)和Iba1(+)细胞。VEGFR1mRNA及其三种配体PlGF、VEGF-A和VEGF-B在各期均有表达，而VEGFR2仅在晚期表达。激光后3d和7d可见PlGF和VEGF蛋白表达。抗VEGFR1(MF1)可抑制视网膜小胶质细胞/巨噬细胞对CNV的浸润，而抗VEGFR2(DC101)对CNV无作用。激光后14天，MF1和DC101抗体均可明显抑制视网膜小胶质细胞/巨噬细胞对CNV的侵袭。因此，VEGFR1和R2在CNV的发病机制中起着不同的作用：在激光后3d，VEGFR1起主导作用；而在激光后14d，两种受体都起着关键作用。体内成像显示，在CX3CR1gfp/GFP和CX3CR1gfp/+小鼠中，表达GFP的小胶质细胞积聚到CNV中。与对照组相比，米诺环素治疗导致CNV前视网膜下间隙凝集素+细胞显著增加，而葡聚糖灌流的新生血管减少。靶向调节视网膜小胶质细胞/巨噬细胞运输的化学吸引分子似乎是控制CNV和治疗湿性AMD的一种引人注目的治疗策略。
Although blocking VEGF has a positive effect in wet age-related macular degeneration (AMD), the effect of blocking its receptors remains unclear. This was an investigation of the effect of VEGF receptor (VEGFR) 1 and/or 2 blockade on retinal microglia/macrophage infiltration in laser-induced choroidal neovascularization (CNV), a model of wet AMD. CNV lesions were isolated by laser capture microdissection at 3, 7, and 14 days after laser and analyzed by RT-PCR and immunofluorescence staining for mRNA and protein expression, respectively. Neutralizing antibodies for VEGFR1 or R2 and the microglia inhibitor minocycline were injected intraperitoneally (IP). Anti-CD11b, CD45 and Iba1 antibodies were used to confirm the cell identity of retinal microglia/macrophage, in the RPE/choroidal flat mounts or retinal cross sections. CD11b(+), CD45(+) or Iba1(+) cells were counted. mRNA of VEGFR1 and its three ligands, PlGF, VEGF-A (VEGF) and VEGF–B, were expressed at all stages, but VEGFR2 were detected only in the late stage. PlGF and VEGF proteins were expressed at 3 and 7 days after laser. Anti-VEGFR1 (MF1) delivered IP 3 days after laser inhibited infiltration of leukocyte populations, largely retinal microglia/macrophage to CNV, while anti-VEGFR2 (DC101) had no effect. At 14 days after laser, both MF1 and DC101 antibodies markedly inhibited retinal microglia/macrophage infiltration into CNV. Therefore, VEGFR1 and R2 play differential roles in the pathogenesis of CNV: VEGFR1 plays a dominant role at 3 days after laser; but both receptors play pivotal roles at 14 days after laser. In vivo imaging demonstrated accumulation of GFP-expressing microglia into CNV in both CX3CR1gfp/gfp and CX3CR1gfp/+ mice. Minocycline treatment caused a significant increase in lectin+ cells in the sub-retinal space anterior to CNV and a decrease in dextran-perfused neovessels compared to controls. Targeting the chemoattractant molecules that regulate trafficking of retinal microglia/macrophage appears to be a compelling therapeutic strategy to control CNV and treat wet AMD.
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影响因子：
3.7
作者：
Huang H;Shen J;Vinores SA
通讯作者：
Vinores SA
影响因子：
4.4
作者：
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