Macrophages promote vasculogenesis of retinal neovascularization in an oxygen-induced retinopathy model in mice
Macrophages promote vasculogenesis of retinal neovascularization in an oxygen-induced retinopathy model in mice
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巨噬细胞促进小鼠氧诱导视网膜病变模型中视网膜新生血管的发生
DOI：
10.1007/s00441-015-2353-y
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发表时间：
2016-06-01
影响因子：
3.6
通讯作者：
Zhang, Jian
中科院分区：
文献类型：
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作者：
Gao, Xiang;Wang, Yu-Sheng;Zhang, Jian
为探讨巨噬细胞在氧诱导的小鼠视网膜新生血管（NV）中的作用，特别是骨髓源性细胞（BMC）的参与及其机制，将绿色荧光蛋白（GFP）转基因小鼠的BMC移植到出生后1天（P）照射的小鼠体内。从P7至P12将小鼠暴露于75%的氧气以引发氧诱导的视网膜病变（OIR）。通过腹腔注射氯膦酸盐脂质体（lip）消耗巨噬细胞。在P12和P17时采集眼睛。进行视网膜平片和组织病理学横截面以分析视网膜NV和BMC募集的严重程度。使用共聚焦显微镜检测CD 31（PECAM-1;内皮细胞标志物）和α-SMA（平滑肌细胞标志物）抗原免疫阳性的BMCs。RT-PCR检测血管内皮生长因子（VEGF）和基质细胞衍生因子-1（SDF-1）mRNA的表达。Western blot检测VEGF、SDF-1、CXCR 4和CD 45蛋白表达。在使用氯膦酸盐-唇进行两次巨噬细胞耗竭（减少38.27 ± 1.92%）后，P12时OIR小鼠的视网膜无血管面积未受影响。在巨噬细胞耗竭4次后，P17时的视网膜无血管面积和NV面积减少（减少79.53 ± 1.02%）;这些发现得到了视网膜平片和组织病理学横截面的支持。在P17时，巨噬细胞耗竭导致BMC募集到NV簇中的显著抑制，视网膜VEGF、SDF-1、CXCR 4和CD 45的表达降低。募集的BMCs主要分化为CD 31阳性的内皮细胞（ECs）和α-SMA阳性的平滑肌细胞（SMCs）。这项研究表明，巨噬细胞促进视网膜NV的血管生成，特别是在小鼠OIR模型中的BMC的贡献，这可能是由VEGF和SDF-1的产生触发的。
To investigate the role of macrophages in oxygen-induced retinal neovascularization (NV) in mice, particularly the involvement of bone marrow-derived cells (BMCs) and the underlying mechanisms, BMCs from green fluorescent protein (GFP) transgenic mice were transplanted into postnatal day (P) 1 mice after irradiation. The mice were exposed to 75 % oxygen from P7 to P12 to initiate oxygen-induced retinopathy (OIR). The macrophages were depleted by injection of clodronate-liposomes (lip) intraperitoneally. The eyes were collected at P12 and P17. Retinal flatmounts and histopathological cross-sections were performed to analyze the severity of retinal NV and BMC recruitment. BMCs immunopositive for CD31 (PECAM-1; endothelial cell marker) and α-SMA (smooth muscle cell marker) antigens were detected using a confocal microscope. Expression of vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1) mRNA was detected by RT-PCR. The VEGF, SDF-1, CXCR4 and CD45 protein expression was detected by western blot examination. The retinal avascular area in OIR mice at P12 was unaffected after macrophage depletion carried out twice (38.27 ± 1.92 % reduction) using clodronate-lip. The retinal avascular area and the NV area at P17 were reduced after macrophage depletion four times (79.53 ± 1.02 % reduction); these findings were supported by retinal flatmounts and histopathological cross-sections. Macrophage depletion led to significant inhibition of BMC recruitment into the NV tufts at P17, with decreased expression of retinal VEGF, SDF-1, CXCR4 and CD45. The recruited BMCs differentiated primarily into CD31-positive endothelial cells (ECs) and α-SMA-positive smooth muscle cells (SMCs). This study suggested that macrophages promoted the vasculogenesis of retinal NV, particularly the contribution of BMCs in the mouse OIR model, which might be triggered by VEGF and SDF-1 production.