树枝状大分子N-乙酰半胱氨酸纳米聚合物对视网膜血管重建的作用  

Effects of nano-polymer dendrimer-N-acetyl cysteine on retinal revascularization

作　　者：杨静 齐赟 YANG Jing;QI Yun(Department of Health Medicine,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,China;Department of Ophthalmology,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,China)

机构地区：[1]西安交通大学第一附属医院健康医学科,陕西省西安市710061 [2]西安交通大学第一附属医院眼科,陕西省西安市710061

出　　处：《眼科新进展》2024年第7期526-530,共5页Recent Advances in Ophthalmology

基　　金：陕西省自然科学基础研究计划(编号:2022JQ-937)。

摘　　要：目的通过对氧诱导的视网膜病变(OIR)小鼠模型抑制活性氧(ROS)的表达,研究ROS抑制剂纳米试剂树枝状大分子(Dendrimer)和N-乙酰半胱氨酸(NAC)的聚合物(D-NAC)对视网膜血管重建的作用及其机制。方法C57BL/6J小鼠随机分为3组:正常对照组、OIR+Dendrimer组(OIR模型+玻璃体内注射Dendrimer)和OIR+D\|NAC组(OIR模型+玻璃体内注射D-NAC),通过从出生后第7天(P7)到P12暴露在含体积分数75%O 2的环境,从P12到P17返回到含体积分数21%O 2的环境,建立小鼠OIR模型。谷胱甘肽(GSH)检测试剂盒检测各组小鼠视网膜GSH浓度。视网膜荧光染色定量对比两OIR组小鼠间视网膜血管闭塞区(VO)和病理性新生血管(NV)面积,qRT-PCR检测两OIR组小鼠之间视网膜NV相关因子的mRNA水平差异,ELISA法检测两OIR组小鼠之间视网膜血管内皮生长因子(VEGF)蛋白表达的差异,qRT-PCR检测两OIR组小鼠之间视网膜Delta样4配体(DLL4)/Notch通路相关因子的mRNA水平差异。结果GSH检测结果显示,P12、P13、P14、P16、P17时,正常对照组与OIR+Dendrimer组小鼠视网膜GSH水平相比,差异均无统计学意义(均为P>0.05);在P16和P17,与OIR+Dendrimer组相比,OIR+D-NAC组小鼠视网膜GSH水平升高(P<0.05)。视网膜铺片染色结果显示,在P17,与OIR+Dendrimer组相比,OIR+D-NAC组VO及病理性NV面积均减少(均为P<0.05)。qRT-PCR结果显示,在P16,与OIR+Dendrimer组相比,OIR+D-NAC组小鼠视网膜成纤维细胞生长因子、促血管生成素、VEGF、VEGF受体2、促红细胞生成素mRNA水平均降低(均为P<0.05);ELISA检测结果显示,与OIR+Dendrimer组相比,OIR+D-NAC组小鼠视网膜VEGF蛋白表达水平降低(P<0.01)。qRT-PCR检测结果显示,在P16,与OIR+Dendrimer组相比,OIR+D-NAC组小鼠视网膜DLL4/Notch通路相关因子Hey1、NRARP、DLL4、Notch1、Hes1和Hey2 mRNA水平均降低(均为P<0.01)。结论ROS抑制剂D-NAC通过抑制DLL4/Notch信号通路促进OIR的生理性NV形成,抑制病理性NV形成的过程。Objective To explore the effects of the reactive oxygen species(ROS)inhibitor Dendrimer and N-acetyl cysteine(NAC)polymer(D-NAC)on retinal revascularization and its mechanism by inhibiting the expression of ROS in mice with oxygen-induced retinopathy(OIR).Methods The C57BL/6J mice were randomly divided into three groups:normal control group(NC group),OIR+Dendrimer group(OIR models+intravitreal injection of Dendrimer),and OIR+D-NAC group(OIR models+intravitreal injection of D-NAC).The mice were exposed to 75%oxygen from postnatal day 7(P7)to P12 and then to 21%oxygen from P12 to P17 to establish OIR models.The concentration of glutathione(GSH)was detected by GSH assay.The areas of retinal vessel occlusion(VO)and pathological neovascularization(NV)were quantitatively compared between the two OIR groups after fluorescence staining of the retina.The mRNA levels of retinal NV-related factors in mice between the two OIR groups were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR).The protein expression of retinal vascular endothelial growth factor(VEGF)in mice between the two OIR groups was detected by enzyme-linked immunosorbent assay(ELISA).The mRNA levels of Delta-like ligand 4(DLL4)/Notch pathway-associated factors in mice between the two OIR groups were detected by qRT-PCR.Results GSH assay results showed that there was no statistically significant difference in the retinal GSH level of mice between the NC group and the OIR+Dendrimer group on P12,P13,P14,P16,and P17(all P>0.05).Compared with the OIR+Dendrimer group,the retinal GSH level of mice in the OIR+D-NAC group increased on P16 and P17(both P<0.05).The retinal flat-mount staining results showed that compared with the OIR+Dendrimer group,the areas of VO and pathological NV in the OIR+D-NAC group decreased on P17(both P<0.05).qRT-PCR results showed that on P16,compared with the OIR+Dendrimer group,the mRNA levels of fibroblast growth factor,angiogenin,VEGF,VEGF receptor 2 and erythropoietin decreased in the OIR+D-NAC group(all

关 键 词：树枝状大分子 N-乙酰半胱氨酸 氧诱导的视网膜病变 血管重建 新生血管 DLL4/Notch信号通路 

分 类 号：R774[医药卫生—眼科]