Technetium-99 Conjugated with Methylene Diphosphonate (99Tc-MDP) Inhibits Experimental Choroidal Neovascularization In Vivo and VEGF-Induced Cell Migration and Tube Formation In Vitro
Technetium-99 Conjugated with Methylene Diphosphonate (99Tc-MDP) Inhibits Experimental Choroidal Neovascularization In Vivo and VEGF-Induced Cell Migration and Tube Formation In Vitro
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DOI：
10.1167/iovs.10-6370
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Ye, Haiyun
中科院分区：
文献类型：
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作者：
Lai, Kunbei;Xu, Li;Ye, Haiyun
目的.目的探讨99 mTc-MDP的衰变产物Tc-99-MDP对脉络膜新生血管（CNV）形成的影响及其机制。C57 BL/6 J小鼠通过激光光凝诱导CNV。在光凝后，每天腹腔内注射0.5 × 10（1）、1 × 10（-1）和2 × 10（-1）μ g/kg剂量的Tc-99-MDP或相同体积的PBS，直至实验结束。激光损伤后7天，用荧光素标记的葡聚糖灌注小鼠，并测量CNV的面积。激光光凝后3 d检测RPE-脉络膜复合体中浸润的巨噬细胞数量、VEGF蛋白水平以及炎症相关分子包括细胞间粘附分子（ICAM）-1、肿瘤坏死因子（TNF）-α和基质金属蛋白酶（MMPs）。还研究了Tc-99-MDP对VEGF诱导的内皮细胞迁移和管形成的影响。对Tc-99-MDP进行了体内和体外毒性评价。与PBS处理相比，Tc-99-MDP处理以剂量依赖性方式显著抑制CNV面积，对视网膜无毒性：0.5 x 10（-1）μ g/kg Tc-99-MDP处理（5698.60 +/- 1037.70 mu m（2）），1 x 10（-1）mu g/kg（3678.34 +/- 1328.18 mu m（2））和2 x 10（-1）mu g/kg 2365.78 ± 923.80 μ m2）对CNV的抑制率分别为36.12%、58.76%和73.48（8920.36 ± 1097.29 μ m（2）; P < 0.001）。Tc-99-MDP治疗导致巨噬细胞浸润到CNV的显著抑制以及VEGF、ICAM-1、TNF-α和MMP-2的蛋白表达下调。Tc-99-MDP对VEGF诱导的内皮细胞增殖、迁移及毛细血管样管形成有抑制作用。用Tc-99-MDP进行抗炎治疗对CNV相关疾病具有治疗潜力。（Invest Ophthal mol维斯科学。2011;52：5702-5712）DOI：10.1167/iovs.10-6370
PURPOSE. To investigate the effects of Tc-99-MDP, a decay product of (99)mTc-MDP, on the development of choroidal neovascularization (CNV), together with its underlying mechanisms.METHODS. C57BL/6J mice were used to induce CNV by laser photocoagulation. Tc-99-MDP at the doses of 0.5 x 10 (1), 1 x 10(-1), and 2 x 10(-1) mu g/kg or the same volume of PBS was intraperitoneally injected daily after photocoagulation until the end of the experiment. Seven days after laser injury, mice were perfused with fluorescein-labeled dextran, and areas of CNV were measured. Numbers of infiltrating macrophages, protein levels of VEGF, and inflammation-related molecules including intercellular adhesion molecule (ICAM)-1, tumor necrosis factor (TNF)-alpha, and matrix metalloproteinases (MMPs) in the RPE-choroid complex were detected 3 days after laser photocoagulation. Effects of Tc-99-MDP on VEGF-induced endothelial cell migration and tube formation were also studied. Toxicity of Tc-99-MDP was evaluated in vivo and in vitro.RESULTS. Areas of CNV were significantly suppressed by Tc-99-MDP treatment without toxicity to the retina compared with PBS treatment in a dose-dependent manner: Tc-99-MDP treatment of 0.5 x 10(-1) mu g/kg (5698.60 +/- 1037.70 mu m(2)), 1 x 10(-1) mu g/kg (3678.34 +/- 1328.18 mu m(2)), and 2 x 10(-1) mu g/kg (2365.78 +/- 923.80 mu m(2)) suppressed the development of CNV by 36.12%, 58.76%, and 73.48%, respectively, compared with that in the PBS treatment group (8920.36 +/- 1097.29 mu m(2); P < 0.001). Tc-99-MDP treatment led to significant inhibition of macrophages infiltrating to CNV together with downregulated protein expressions of VEGF, ICAM-1, TNF-alpha, and MMP-2. Tc-99-MDP also showed an inhibitive effect on cell proliferation and VEGF-induced migration and capillary-like tube formation of endothelial cells.CONCLUSIONS. Anti-inflammatory treatment with Tc-99-MDP has therapeutic potential for CNV-related diseases. (Invest Ophthal mol Vis Sci. 2011;52:5702-5712) DOI:10.1167/iovs.10-6370