[1]杨静,齐赟.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[J].眼科新进展,2024,44(7):526-530.[doi:10.13389/j.cnki.rao.2024.0101]
 YANG Jing,QI Yun.Effects of nano-polymer dendrimer-N-acetyl cysteine on retinal revascularization[J].Recent Advances in Ophthalmology,2024,44(7):526-530.[doi:10.13389/j.cnki.rao.2024.0101]
点击复制

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/HTML
分享到:

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
44卷
期数:
2024年7期
页码:
526-530
栏目:
实验研究
出版日期:
2024-07-01

文章信息/Info

Title:
Effects of nano-polymer dendrimer-N-acetyl cysteine on retinal revascularization
作者:
杨静齐赟
710061 陕西省西安市,西安交通大学第一附属医院健康医学科(杨静);710061 陕西省西安市,西安交通大学第一附属医院眼科(齐赟)
Author(s):
YANG Jing1QI Yun2
1.Department of Health Medicine,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,China
2.Department of Ophthalmology,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,Shaanxi Province,China
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
Keywords:
dendrimer N-acetyl cysteine oxygen-induced retinopathy revascularization neovascularization Delta-like ligand 4/Notch signaling pathway
分类号:
R774
DOI:
10.13389/j.cnki.rao.2024.0101
文献标志码:
A
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% O2的环境,从P12到P17返回到含体积分数21% O2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。
Abstract:
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 P<0.05); ELISA results showed that compared with the OIR+Dendrimer group, the protein expression of VEGF decreased in the OIR+D-NAC group (P<0.01). qRT-PCR results showed that compared with the OIR+Dendrimer group, the mRNA levels of DLL4/Notch pathway-associated factors such as Hey1, NRARP, DLL4, Notch1, Hes1 and Hey2 decreased on P16 in the OIR+D-NAC group (all P<0.01).
Conclusion ROS inhibitor D-NAC promotes physiological NV formation in OIR by inhibiting the DLL4/Notch signaling pathway, and restrains the pathological NV formation.

参考文献/References:

[1] CHAN T C,WILKINSON BERKA J L,DELIYANTI D,HUNTER D,FUNG A,LIEW G,et al.The role of reactive oxygen species in the pathogenesis and treatment of retinal diseases[J].Exp Eye Res,2020,201:108255.
[2] FENG Y,GROSS S,CHATTERJEE A,WANG Y,LIN J,HAMMES H P.Transcription of inflammatory cytokine TNF-α is upregulated in retinal angiogenesis under hyperoxia[J].Cell Physiol Biochem,2016,39(2):573-583.
[3] RIVERA J C,SITARAS N,NOUEIHED B,HAMEL D,MADAAN A,ZHOU T,et al.Microglia and interleukin-1β in ischemic retinopathy elicit microvascular degeneration through neuronal semaphorin-3A[J].Arterioscler Thromb Vasc Biol,2013,33(8):1881-1891.
[4] MANEA S A,CONSTANTIN A,MANDA G,SASSON S,MANEA A.Regulation of Nox enzymes expression in vascular pathophysiology:focusing on transcription factors and epigenetic mechanisms[J].Redox Biol,2015,5:358-366.
[5] AL-SHABRAWEY M,BARTOLI M,EL-REMESSY A B,MA G,MATRAGOON S,LEMTALSI T,et al.Role of NADPH oxidase and Stat3 in statin-mediated protection against diabetic retinopathy[J].Invest Ophthalmol Vis Sci,2008,49(7):3231-3238.
[6] WANG J,LI M,GENG Z,KHATTAK S,JI X,WU D,et al.Role of oxidative stress in retinal disease and the early intervention strategies:a review[J].Oxid Med Cell Longev,2022,2022:7836828.
[7] WILKINSON-BERKA J L,RANA I,ARMANI R,AGROTIS A.Reactive oxygen species,Nox and angiotensin II in angiogenesis:implications for retinopathy[J].Clin Sci (Lond),2013,124(10):597-615.
[8] PARAD R B,ALLRED E N,ROSENFELD W N,DAVIS J M.Reduction of retinopathy of prematurity in extremely low gestational age newborns treated with recombinant human Cu/Zn superoxide dismutase[J].Neonatology,2012,102(2):139-144.
[9] PENN J S,TOLMAN B L,BULLARD L E.Effect of a water-soluble vitamin E analog,trolox C,on retinal vascular development in an animal model of retinopathy of prematurity[J].Free Radic Biol Med,1997,22(6):977-984.
[10] ALDINI G,ALTOMARE A,BARON G,VISTOLI G,CARINI M,BORSANI L,et al.N-Acetylcysteine as an antioxidant and disulphide breaking agent:the reasons why[J].Free Radic Res,2018,52(7):751-762.
[11] SHARMA R,SHARMA A,KAMBHAMPATI S P,REDDY R R,ZHANG Z,CLELAND J L,et al.Scalable synthesis and validation of PAMAM dendrimer-N-acetyl cysteine conjugate for potential translation[J].Bioeng Transl Med,2018,3(2):87-101.
[12] ZHOU B,LIN W,LONG Y,YANG Y,ZHANG H,WU K,et al.Notch signaling pathway:architecture,disease,and therapeutics[J].Signal Transduct Target Ther,2022,7(1):95.
[13] LOBOV I,MIKHAILOVA N.The role of Dll4/Notch signaling in normal and pathological ocular angiogenesis:Dll4 controls blood vessel sprouting and vessel remodeling in normal and pathological conditions[J].J Ophthalmol,2018,2018:3565292.
[14] CONNOR K M,KRAH N M,DENNISON R J,ADERMAN C M,CHEN J,GUERIN K I,et al.Quantification of oxygen-induced retinopathy in the mouse:a model of vessel loss,vessel regrowth and pathological angiogenesis[J].Nat Protoc,2009,4(11):1565-1573.
[15] PITHA I,KAMBHAMPATI S,SHARMA A,SHARMA R,MCCREA L,MOZZER A,et al.Targeted microglial attenuation through dendrimer-drug conjugates improves glaucoma neuroprotection[J].Biomacromolecules,2023,24(3):1355-1365.
[16] SAITO Y,GEISEN P,UPPAL A,HARTNETT M E.Inhibition of NAD(P)H oxidase reduces apoptosis and avascular retina in an animal model of retinopathy of prematurity[J].Mol Vis,2007,13:840-853.
[17] AIELLO L P.Angiogenic pathways in diabetic retinopathy[J].N Engl J Med,2005,353(8):839-841.
[18] CHAN E C,VAN WIJNGAARDEN P,LIU G S,JIANG F,PESHAVARIYA H,DUSTING G J.Involvement of Nox2 NADPH oxidase in retinal neovascularization[J].Invest Ophthalmol Vis Sci,2013,54(10):7061-7067.
[19] HELLSTRM M,PHNG L K,HOFMANN J J,WALLGARD E,COULTAS L,LINDBLOM P,et al.Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis[J].Nature,2007,445(7129):776-780.
[20] PITULESCU M E,SCHMIDT I,GIAIMO B D,ANTOINE T,BERKENFELD F,FERRANTE F,et al.Dll4 and Notch signalling couples sprouting angiogenesis and artery formation[J].Nat Cell Biol,2017,19(8):915-927.

相似文献/References:

[1]郝瑞霖,杨炜,张奕霞,等.乙酰半胱氨酸对角膜新生血管形成过程中瘦素和PEDF表达的调节[J].眼科新进展,2014,34(8):718.[doi:10.13389/j.cnki.rao.2014.0196]
 HAO Rui-Lin,YANG Wei,ZHANG Yi-Xia,et al.Regulative effects of acetylcysteine on leptin and PEDF in corneal neovascularization[J].Recent Advances in Ophthalmology,2014,34(7):718.[doi:10.13389/j.cnki.rao.2014.0196]

备注/Memo

备注/Memo:
陕西省自然科学基础研究计划(编号: 2022JQ-937)
更新日期/Last Update: 2024-07-05