摘要
目的建立氧诱导新生小鼠视网膜病变(oxygen-induced retinopathy,OIR)模型,观察视网膜血管新生情况,检测OIR病变过程中调控血管新生和氧应激相关基因表达的变化。方法新生小鼠出生后d 7,置75.5%高氧环境中饲养5 d,再返回正常氧环境继续饲养5 d,建立OIR模型。运用视网膜免疫荧光染色检测OIR小鼠视网膜中血管新生情况;运用Real-time PCR技术检测基因表达情况。结果视网膜免疫荧光染色结果显示,OIR小鼠视网膜在高氧诱导视网膜微血管消退的基础上,相对缺氧状态诱导出现明显的新生血管。Real-time PCR结果显示OIR模型小鼠视网膜中:血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)及其受体(VEGFR)家族中:VEGFA、VEGFD、VEGFR1、VEGFR2的基因表达分别上调1.73、1.71、1.48、1.80倍;血小板衍生生长因子(platelet-derived growth factor,PDGF)及其受体(PDGFR)家族中:PDGFA、PDGFB、PDGFRa、PDGFRb的基因表达分别上调1.29、1.71、1.42、1.42倍;基质金属蛋白酶(matrix metalloproteinases,MMPs)中的MMP2基因表达上调1.48倍;参与调控氧化应激的核转录因子Nrf2[nuclear factor(erythroid-derived 2)-like 2]及受其调控的下游基因包括谷氨酸-半胱氨酸连接酶(glutamatecysteine ligase,GCL)的催化(GCLC)和调节(GCLM)亚单位的基因表达均下调,表达量分别为正常的0.28、0.76、0.49。结论 OIR小鼠视网膜病变过程中,视网膜中促进血管新生相关基因表达上调,而抗氧化相关基因表达下调。
Aim To observe the retinal angiogenesis and detect the altered expression of genes related with angiogenesis and oxidative stress during the develop-ment of oxygen-induced retinopathy ( OIR) in newborn mice. Methods OIR was established in newborn mice according to the protocol of Smith et al. Newborn mice at 7 days old were placed into 75 . 5% oxygen for up to 5 days, and then they were put in room air for another 5 days. Retinal neovascularization was ob-served by immunofluorescence staining with cluster of differentiation 31 ( CD31 ) . Gene expression was de-tected using Real-time PCR analysis. Retinal CD31 immunofluorescence staining assay showed that relative hypoxia induced retinal neovascularization in OIR mice after hyperoxia-induced subside of retinal microvascu-lar. Results Real-time PCR analysis showed that vas-cular endothelial growth factor ( VEGF) and its recep-tor ( VEGFR) such as VEGFA, VEGFD, VEGFR1, VEGFR2 gene expression were increased in OIR mouseas compared to control. Platelet-derived growth factor ( PDGF) and its receptor ( PDGFR) such as PDGFA, PDGFB, PDGFRa, PDGFRb gene expression was also increased in OIR mouse as compared to control. Matrix metalloproteinases ( MMPs ) such as MMP2 gene ex-pression were increased in OIR mouse as compared to control. Gene expressions of nuclear factor-related fac-tor ( Nrf2 ) and its downstream genes such as the two subunits of glutamate-cysteine ligase ( GCL):the cata-lytic subunit ( GCLC) and regulatory subunit ( GCLM) were both decreased in OIR mouse as compared to con-trol. Conclusion Our research demonstrates that the expression of genes related with angiogenesis is in-creased in retinas in the development of OIR in mice, whereas the expression of Nrf2 and its downstream genes is all decreased.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2014年第10期1397-1401,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81173517)
上海市卫生局科研基金资助项目(No 2012-4140)