玻璃体腔注射VAS2870对小鼠氧诱导视网膜病变的保护作用

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C57 BL/6 J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7 d ( 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 mRNA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 mRNA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 mRNA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。

Protection of VAS2870 to oxygen-induced retinopathy in mice

?AIM:To investigate the protective effect of VAS2870 on mice oxygen - induced retinopathy ( OIR ) and the possible underlying mechanisms.? METHODS: Neonatal C57BL/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 from the postnatal 7d ( P7 ) to the postnatal 12d ( P12 ) to induced OIR. VAS2870 was administered by intravitreal injection (0. 5μL) 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 RT-PCR, 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 VAS2870
has potent anti-oxidative stress effects in the retina, indicating its potential as a treatment for retinopathy of prematurity.