玻璃体腔注射VAS2870对小鼠氧诱导视网膜病变的保护作用

Protection of VAS2870 to oxygen-induced retinopathy in mice

目的:观察玻璃体腔注射VAS2870对C57小鼠氧诱导视网膜病变的影响。方法:将新生C57BL/6J小鼠随机分为3组,分别为正常对照组、VAS2870注射OIR组和无菌PBS缓冲液注射OIR组。将后两组小鼠在出生后第7d(P7)至P12置于体积分数为75%±2%的恒定高氧氧箱中以构建OIR模型,在P12时给予幼鼠双眼玻璃体腔注射VAS2870(0.5μL),另一组幼鼠双眼注射同等剂量的无菌PBS缓冲液。三组小鼠均在P17时取右眼行视网膜铺片和Lectin染色,观察视网膜中央无血管区及病理性新生血管的情况;取右眼行视网膜组织定量检测ROS/RNS含量;取左眼应用RT-PCR检测Nox4 m RNA含量,并应用Western-blot测定视网膜组织中VEGF的表达。结果:VAS2870注射OIR组小鼠视网膜中央无血管区面积较无菌PBS缓冲液注射OIR组明显减少(P<0.05),病理性新生血管数目明显减少(P<0.05);VAS2870注射OIR组小鼠视网膜组织Nox4 m RNA的表达量明显低于无菌PBS缓冲液注射组;VAS2870注射OIR组小鼠视网膜组织ROS含量较无菌PBS缓冲液注射组明显降低(P<0.05);VAS2870注射OIR组小鼠视网膜组织VEGF的表达明显低于无菌PBS缓冲液注射组(P<0.05)。结论:在小鼠OIR模型中,VAS2870可抑制Nox4 m RNA的表达,减少ROS/RNS,下调VEGF的生成,在视网膜病变进程中具有保护作用。

AIM：To investigate the protective effect of VAS2870 on mice oxygen- induced retinopathy（ OIR） and the possible underlying mechanisms. METHODS： Neonatal C57 BL /6J mice were divided randomly into three groups： normoxic control group,PBS injection of OIR group and VAS2870 injection of OIR group. The mice of the latter two groups were exposed to 75% oxygen fromthe postnatal 7d（ P7） to the postnatal 12d（ P12） to induced OIR. VAS2870 was administered by intravitreal injection（ 0. 5μ L） in a mice model of OIR in P12. Another set of mice model of OIR were received a similar treatment with PBS. All eyes were collected at P17. The right eyes were whole mounted and stained with Lectin to observe the growth of retinal vessels; The eyes were enucleated to assess the levels of reactive oxygen species ROS / RNS. The expression of vascular endothelial growth factor（ VEGF）and Nox4 mRNA were detected by western blot and RTPCR,respectively.RESULTS： In the retina of OIR,VAS2870 reduced the retinal avascular area and neovascularization, the hypoxia- induced increase in ROS levels, the protein expression of VEGF and gene expression of Nox4 mRNA.CONCLUSION： NOX4 enzyme inhibition with VAS2870has potent anti- oxidative stress effects in the retina,indicating its potential as a treatment for retinopathy of prematurity.