【摘要】 目的:通过观察apelin及其受体APJ和一氧化氮合酶(NOS)在氧诱导的新生小鼠增生性视网膜病变中的表达,探讨apelin/APJ和一氧化氮(NO)是否参与了早产儿视网膜病变新生血管的发生。方法:36只7日龄C57BL/6J新生小鼠随机均分为高氧组和对照组。高氧组暴露在75%±2%的氧浓度下5 d后,在正常空气环境下饲养5 d;对照组小鼠在正常氧环境下饲养10 d。两组均在17日龄处死,取左眼眼球做石蜡切片,HE染色计数突破内界膜的血管内皮细胞核数,以判断造模是否成功。实时荧光定量PCR检测右眼视网膜组织apelin和APJmRNA的表达,免疫荧光组织化学法检测视网膜apelin、APJ、eNOS和iNOS蛋白的表达。结果:高氧组视网膜平均每个切面突破内界膜的血管内皮细胞核数(35.13±10.13)明显高于对照组(0.30±0.21,P<0.01),说明造模成功。高氧组apelin和APJ mRNA水平显著高于对照组,分别为对照组的32.2倍和17.6倍(均P<0.01)。组织免疫荧光结果显示高氧组apelin和APJ蛋白表达明显强于对照组,并且主要强表达于新生血管周围;高氧组eNOS和iNOS蛋白表达明显强于对照组,但主要强表达于新生血管下方视网膜组织,在突破内界膜的血管内皮细胞核周围未见表达明显增强。结论:Apelin/APJ及NOS可能与氧诱导的新生小鼠增生性视网膜病变新生血管的发生有关。更多还原

【Abstract】 AIM:To explore whether apelin and its receptor(APJ) as well as nitric oxide synthase(NOS) were involved in the neovascularization of retinopathy of prematurity by observing their expression characteristics in the retina of a mouse model of oxygen-induced retinopathy.METHODS: Thirty-six 7-day-old C57BL/6J neonatal mice were randomly divided into high-oxygen group and control group.The mice in high-oxygen group were exposed to 75%±2% oxygen for 5 d,and then changed to normoxia for the next 5 d.The control ones were kept in a normoxic environment for 10 d.The mice in both groups were sacrificed on day 17 of life.The left eyes were isolated to make paraffin sections.The number of endothelial cell nuclei extending from the retinal area into the vitreous area was counted under microscope with HE staining to judge whether the model was successfully established,and immunofluorescence was performed to detect the protein expression of apelin,APJ,eNOS and iNOS.The retinas of the right eyes were dissected for applying real-time fluorescence quantitative PCR to detect the mRNA levels of apelin and APJ.RESULTS: The number of neovascular endothelial cell nuclei extending from the retinal area into the vitreous area in high-oxygen group was much higher than that in control group(35.13±10.13 vs 0.30±0.21,P<0.01).The mRNA levels of apelin and APJ were higher than those in control group(P<0.01),by 32.2 times and 17.6 times,respectively.The expression of apelin and APJ proteins was significantly increased in high-oxygen group,mainly around the neovessels.The expression of eNOS and iNOS proteins was also significantly increased in high-oxygen group,mainly in the retinal tissue under the neovessels,but not around the neovascular endothelial cell nuclei.CONCLUSION: Apelin and its receptor as well as NOS may be involved in the neovascularization in oxygen-induced retinopathy.更多还原