MMP-2和VEGF在视网膜新生血管的表达

目的检测基质金属蛋白酶-2(matrix metalloproteinases-2,MMP-2)和血管内皮生长因子(vascular endothelial growthfactor,VEGF)在视网膜新生血管中的表达,探讨两者的相互关系。方法 取7 d龄C57BL/6J小鼠40只,随机分为高氧组和对照组,每组各20只。建立高氧诱导的视网膜新生血管小鼠模型。采用ADP酶组织化学染色、HE染色和免疫组化方法分别观察2组视网膜血管的变化、计数突破视网膜内界膜的内皮细胞核数目和检测MMP-2、VEGF的表达。结果 高氧组视网膜铺片可见大量视网膜新生血管形成,对照组未见新生血管形成;高氧组突破视网膜内界膜内皮细胞核数目为14(14.230±5.388),与对照组数目0(0.110±0.386)相比差异有统计学意义(t=23.537,P<0.001);对照组和高氧组视网膜组织中VEGF的积分光密度值分别为36.81±14.60、60.85±24.55,差异有统计学意义(t=3.348,P<0.01);对照组和高氧组视网膜组织中MMP-2的积分光密度值分别为16.33±4.13、21.12±6.29,差异有统计学意义(t=3.160,P<0.01)。高氧组视网膜组织中MMP-2和VEGF的表达呈正相关(r=0.633,P<0.01)。结论 在ROP小鼠模型的视网膜组织中,VEGF、MMP-2的表达同步升高,二者的高表达可能与视网膜新生血管形成密切相关。     

MMP-2和VEGF在视网膜新生血管中的表达及意义

目的探讨基质金属蛋白酶-2（MMP-2）和血管内皮生长因子（VEGF）在视网膜新生血管中的表达及意义。方法取C57BL/6J小鼠60只,随机分为高氧组和正常组,各30只。以高浓度氧诱导小鼠建立视网膜新生血管模型。采用ADP酶视网膜铺片、苏木精-伊红染色及免疫组织化学法分别观察视网膜血管的改变、计数视网膜新生血管内皮细胞数并检测MMP-2、VEGF蛋白的表达。结果高氧组视网膜可见大量新生血管形成;突破视网膜内界膜的新生血管内皮细胞核数为（33.51±2.55）个,与对照组相比差异有统计学意义（t=9.345,P〈0.05）。高氧组与对照组比较,MMP-2、VEGF蛋白在神经节细胞层、内丛状层、内核层和突破视网膜内界膜的新生血管中高表达,且二者表达呈正相关（r=0.825,P〈0.05）。结论MMP-2、VEGF共同促进视网膜新生血管的形成,且二者可能具有协同作用。     

基质金属蛋白酶抑制剂对视网膜新生血管化抑制作用的实验研究

目的 探讨基质金属蛋白酶（PPMS）抑制剂对视网膜新生血管化抑制作用的应用价值。方法 将新生7天的C57BL／6J小鼠暴露75％高氧中5天,再置于一般空气中5天,造成视网膜新生血管化的动物模型,分成3组,其中1组为对照组。实验组中1组给予Batimastat腹腔注射,另1组注射生理盐水作对照。用免疫组化染色,并用显微镜下微血管计数方法研究视网膜新生血管及其使用MMPS抑制剂的情况。同时采用酶谱法,分析MMPS在各组中含量的变化。结果 注射Batimastat的试验组微血管的数量明显低于注射生理盐水的对照组,同时MMPS的表达也减少,具有统计学显著性差异。结论 Batimastat作为一种新的MMPS抑制可以抑制视网膜新生血管化,并减低MMPS的表达。     

早期糖尿病大鼠视网膜中MMP-3和VEGF表达的意义

目的:探讨基质金属蛋白酶-3(MMP-3)和血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)在早期糖尿病大鼠视网膜的表达及其意义。方法:制备类似人类Ⅱ型糖尿病大鼠动物模型,12wk取眼球、应用免疫组化Envision法检测VEGF和MMP-3在视网膜的表达。结果:MMP-3和VEGF在正常视网膜的染色均为阴性,而在糖尿病大鼠视网膜的内界膜均呈阳性染色反应;VEGF还在视网膜内核层有散在的弱阳性表达。结论:MMP-3和VEGF可能在糖尿病视网膜新生血管的发生过程中起重要作用,且二者之间存在协同关系。     

抗VEGF药物治疗早产儿视网膜病变的研究进展

早产儿视网膜病变(retinopathy of prematurity,ROP)是目前全世界儿童致盲的主要原因之一[1]。以往对于阈值期和高危阈值前期的ROP采用激光或冷凝治疗,因激光和冷凝术均是破坏性的,不可避免地导致周边视野永久丧失,且并发症较多,术后仍有部分患儿病情无法控制,最终视力完全丧失。因此需要寻找新的治疗方法,近年来有较多的临床数据表明,玻璃体内注射抗血管内皮生长因子(anti-vascular endothelial growth factor,anti-VEGF)治疗ROP是一种有效的治疗方法。我们将对抗VEGF药物在ROP治疗的研究进展进行近期的文献综述。     

反义VEGF对视网膜新生血管的干预研究

目的 将反义VEGF质粒用脂质体包裹注入视网膜新生血管模型小鼠眼中，观察其对新生血管的抑制作用。方法 将出生7d的C57BL／6J小鼠44只放入高氧环境中饲养，另外8只于空气环境中饲养。5d后出高氧环境，随机取36只分成3组。将质粒与脂质体以1：5(W／V)的比例混合，室温下静置30min。大剂量组玻璃体腔内注入质粒PCR3．1／Anti—VEGF121 0．085μg；小剂量组玻璃体腔内注入质粒PCR3．1／Anti—VEGF121 0．038μg；PCR3．1组玻璃体腔内注入质粒PCR3．1 0．053μg，之后空气环境饲养5d。解剖镜下视网膜铺片观察视网膜血管的分布情况；组织切片任意选取不包括视乳头的20张切片，计数突出内界膜位于视网膜表面的细胞核数；VEGF免疫组化染色阳性细胞。结果 视网膜铺片原中周部血管紊乱分布部位，血管分布均匀。而对照组(注射PCR3．1)则无变化。内皮细胞计数小剂量组、大剂量组较高氧组、对照组显著减少，在P17的VEGF在毛细血管的血管内皮细胞的表达均有显著降低，但程度上有差异。结论反义VEGF质粒对于视网膜新生血管的产生有抑制作用，在一定程度上减少了视网膜新生血管的产生。对血管内皮细胞表达VEGF有抑制作用。     

PTK787抑制早产儿视网膜病变大鼠模型新生血管的形成

目的探讨受体酪氨酸激酶亚群抑制剂PTK787对早产儿视网膜病变(retinopathy of prematurity,ROP)新生血管形成的抑制作用。方法建立波动氧(体积分数80%和10%氧浓度24h反复交替)诱导的SD大鼠ROP模型。67只新生SD大鼠随机设立对照组(22只)、模型组(22只)、治疗组(23只,腹腔注射PTK78750mg.kg-1);分别于第12天和第17天,每组随机抽取8只新生鼠,一侧眼球采用ADP酶组织化学法进行视网膜铺片,观察视网膜血管改变;另一侧眼球视网膜组织切片观察并计数突破视网膜内界膜的血管内皮细胞核数目。结果波动氧可成功诱导SD新生大鼠ROP模型,PTK787可抑制氧诱导新生鼠视网膜病变模型新生血管的形成。第12天和第17天时,模型组视网膜ADP酶组织化学铺片,均较对照组血管分布、密度改变明显;而治疗组视网膜铺片血管密度较模型组明显下降。第17天突破视网膜内界膜的血管内皮细胞核计数结果显示,给氧模型组31.360±4.543与正常对照组1.700±1.216比较,差异有显著统计学意义(t=-56.414,P<0.001)。治疗组6.800±2.107与模型组相比,血管内皮细胞核数显著减少(t=-43.869,P<0.001)。结论 PTK787可以抑制视网膜新生血管的形成,有望成为治疗ROP的有效途径。     

肝细胞生长因子与视网膜新生血管形成

视网膜新生血管形成可造成眼部多种组织成分的广泛损害,已成为世界范围的致盲性疾病。其发生及发展过程复杂,需要多种因素参与,包括多种生长因子的相互作用等。肝细胞生长因子(HGF)作为多效性生长因子,对新生血管形成的作用逐渐被认识。本文主要探讨HGF在视网膜新生血管发病机制中的作用以及与血管内皮生长因子(VEGF)的关系。     

基质金属蛋白酶与糖尿病视网膜病变新生血管生成

基质金属蛋白酶（MMPS）是新生血管产生的必须条件之一,视网膜新生血管生成是糖尿病视网膜病变增殖期的标志。本文就基质金属蛋白酶的分子生物学特性和其在糖尿病视网膜病变发病过程中的作用作一综述。     

视网膜新生血管动物模型的制备

目的制备视网膜新生血管动物模型,为今后的视网膜新生血管相关疾病的研究提供稳定的模型.方法以出生7d的C57BL/6J小鼠56只,雌雄兼有,随机将28只放入75%±2%高氧环境,控制室温23℃±2℃,日光照明,5d后返回空气环境;另一组28只置于23℃±2℃空气环境中饲养作为对照.随机于出生后12、14、17、21、22、25d取高氧组和空气组小鼠行视网膜铺片、血管内皮生长因子(vascular endothelial growth factor,VEGF)免疫组化染色,观察VEGF的表达情况,并对出生后17d小鼠的视网膜铺片、石蜡切片HE染色、VEGF免疫组化染色.结果高氧诱导组出生后17d视网膜无血管区面积,穿过视网膜内界膜细胞核计数明显高于空气组.血管内皮细胞VEGF的表达从出生后14d开始有阳性表达,阳性表达逐渐增强,出生后17d达到高峰,之后逐渐下降,持续至出生后21d.结论该模型为一种合适的视网膜新生血管动物模型.     

Inhibition of LY294002 in retinal neovascularization via down-regulation the PI3K/AKT-VEGF pathway in vivo and in vitro

AIM: To investigate the effects of the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 on retinal neovascularization (RNV) in the oxygen-induced retinopathy (OIR) mouse model and human umbilical vein endothelial cells (HUVECs). METHODS: C57BL/6J mice were randomly divided into normoxia-control, OIR-control and LY294002 treatment groups. LY294002 or phosphate-buffered solution was intraperitoneally injected daily into mouse pups from P6 to P9 in LY294002 treatment group or OIR-control group. Morphological and pathological changes in RNV, as well as expression levels of PI3K, serine-threonine kinase (AKT) and vascular endothelial growth factor (VEGF) were observed. HUVECs treating with LY294002 were exposed to hypoxia; the expression of PI3K, AKT and VEGF were examined by Western blot and RT-PCR analyses. RESULTS: Compared with the OIR-control group, LY294002 significantly inhibit RNV. Adenosine diphosphatase (ADPase) staining and hematoxylin and eosin staining indicated that the clock hour scores of neovascularization and the nuclei of pre-retinal neovascular cells in the LY294002 treatment group were clearly less than those in the OIR-control group (1.41±0.52 vs 6.20±1.21; 10.50±1.58 vs 22.25±1.82, both P<0.05). Intravitreal injection of LY294002 (in the LY294002 treatment group) markedly decreased PI3K/AKT-VEGF expression compared with the OIR-control group by immunohistochemistry, Western blotting and RT-PCR (all P<0.05). In HUVECs treated with hypoxia, expression of PI3K, AKT and VEGF were downregulated in the hypoxia-LY294002 group (all P<0.05). CONCLUSION: The PI3K inhibitor LY294002 can inhibit RNV by downregulating PI3K, AKT, and VEGF expression in vivo and in vitro. LY294002 may provide an effective method for preventing retinopathy of prematurity (ROP).     

CCR7/p-ERK1/2/VEGF signaling promotes retinal neovascularization in a mouse model of oxygen-induced retinopathy

AIM: To investigate the role of CCR7/p-ERK1/2/VEGF signaling in the mouse model of oxygen-induced retinopathy (OIR). METHODS: Neonatal C57BL/6J mice were evenly randomized into four groups: normoxia, OIR, OIR control (treated with scramble siRNA), and OIR treated (treated with CCR7 siRNA). Normoxia group was not specially handled. Postnatal day 7 (P7) mice in the OIR group were exposed to 75%±5% oxygen for 5d (P7-P12) and then maintained under normoxic conditions for 5d (P12-P17). Mice in the OIR control and OIR treated groups were given injections of scramble or CCR7 siRNA plasmid on P12 before returning to normoxic conditions for 5d (P12-P17). Retina samples were collected from all mice on P17, stained with adenosine diphosphatase (ADPase), and retinal neovascularization (RNV) was assessed. Retinas were also stained with hematoxylin and eosin (H&E) for RNV quantitation. The distribution and expression of CCR7, p-ERK1/2 and vascular endothelial growth factor (VEGF) were assessed via immunohistochemistry, Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: High oxygen promoted retinal neovascularization (P<0.05) and increased the number of endothelial nuclei in new vessels extending from the retina to the vitreous body; CCR7 promoted this process (P<0.05). CCR7 and VEGF mRNA were expressed at higher levels in the OIR and OIR control groups than in the normoxia and OIR treated groups. CCR7, p-ERK1/2, and VEGF protein were expressed in the retinas of mice in the OIR and OIR control groups. Intravitreal injection of CCR7 siRNA significantly reduced CCR7, p-ERK1/2, and VEGF expression in the OIR mouse model (all P<0.05). CCR7 significantly enhanced the neovascularization and non-perfusion areas in the OIR group (P<0.05). CCR7 siRNA significantly reduced levels of p-ERK1/2 and VEGF as compared to OIR controls (P<0.05). CONCLUSION: These results suggest that CCR7/p-ERK 1/2/VEGF signaling plays an important role in OIR. CCR7 may be a potential target for the prevention and treatment of retinopathy of prematurity.     

Matrix metalloproteinase-9 and vascular endothelial growth factor expression change in experimental retinal neovascularization

AIM: To investigate the signal transduction mechanism of matrix metalloproteinase-9 (MMP-9) mediated- vascular endothelial growth factor (VEGF) expression and retinal neovascularization (RNV) in oxygen-induced retinopathy (OIR) model. METHODS: C57BL/6J mice were divided into four groups: control group, OIR group, OIR control group (phosphate-buffered saline by intravitreal injection) and treated group [tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) by intravitreal injection]. OIR model was established in C57BL/6J mice exposed to 75%±2% oxygen for 5d. mRNA level and protein expression of MMP-9, TIMP-1 and VEGF were measured by real-time polymerase chain reaction and Western blotting, and located by immunohistochemistry. RESULTS: Levels of MMP-9 and VEGF in retina were significantly increased in animals with OIR and OIR control group. Levels of TIMP-1 in retina was significantly reduced in animals with OIR and OIR control group. Furthermore, a significant correlation was found between MMP-9 and VEGF. Intravitreal injection of TIMP-1 significantly reduced MMP-9 and VEGF expression of the OIR mouse model (all P<0.05). CONCLUSION: These results demonstrate that MMP-9-mediated up-regulation of VEGF promotes RNV in retinopathy of prematurity (ROP). TIMP-1 may be a potential target for the prevention and treatment of ROP.     

可溶性VEGF受体1在视网膜新生血管疾病中的研究进展

视网膜新生血管性疾病的共同特征是病理性新生血管形成。目前研究的内源性视网膜新生血管因子最主要的是VEGF。可溶性VEGF受体1(sFlt-1)是VEGFR-1的mRNA胞外区剪接形成的可溶性形式,只编码胞外区,缺乏细胞内酪氨酸激酶区域,所以其仅具有与配体结合的能力,而无信号转导能力,从而阻止新生血管的形成。sFlt-1作为近年来的研究热点,有可能成为治疗该类疾病的新的基因治疗方法。本文就sFlt-1在视网膜新生血管疾病治疗中的作用机制及研究进展做一综述。     

多巴胺对氧诱导视网膜病变小鼠视网膜新生血管的影响

目的 观察多巴胺(DA)及多巴胺D2受体(DRD2)在氧诱导视网膜病变(OIR)小鼠模型中对视网膜新生血管的作用。方法 随机将180只小鼠分为空白组、OIR组、NaCl组、DA组、Quinpirole(DRD2激动剂)组、Spiperone(DRD2抑制剂)组,每组30只。空白组小鼠在正常环境中饲养,其他各组小鼠在7 d龄时于高氧环境中饲养5 d,于小鼠12 d龄时返回正常环境中,OIR组不做药物干预,NaCl组、DA组、Quinpirole组分别于小鼠右眼玻璃体内注射8.5 g·L^-1 NaCl、15 mmol·L^-1的DA溶液、602μmol·L^-1的Quinpirole溶液,Spiperone组小鼠右眼玻璃体内依次注射154μmol·L^-1的Spiperone溶液和15 mmol·L^-1的DA溶液各1μL。小鼠17 d龄时处死,摘取右侧眼球进行后续实验。采用苏木精-伊红(HE)染色和视网膜铺片检查各组小鼠视网膜新生血管情况;采用Q-PCR和Western blot检测各组...     

氧诱导小鼠视网膜新生血管的实验研究

目的 制作氧诱导视网膜新生血管的小鼠动物模型并了解其视网膜血管内皮生长因子(VEGF)的变化.方法将出生后第7 d的C57BL/6J小鼠置于75%的高氧环境中5 d,再置于普通空气中5 d.在空气中饲养的小鼠为对照组.两组进行视网膜铺片,ADPase染色,组织切片染色,ELISA测定视网膜VEGF蛋白含量.结果实验组新生血管形成率为100%,对照组未见新生血管.实验组小鼠生后第17 d时突破视网膜内界膜的血管内皮细胞核数目达(46.7±11.1)个,对照组不足2个.第12 d实验组视网膜VEGF蛋白水平比对照组下降,第17 d比对照组升高. 结论该动物模型是研究早产儿视网膜病变(ROP)发病机制及治疗的合适模型.VEGF是造成视网膜新生血管发生的主要机制之一.     

玻璃体内注射贝伐单抗治疗早产儿视网膜病变的临床进展

血管内皮细胞生长因子(VEGF)在早产儿视网膜病变(ROP)的发生过程中发挥重要作用.在ROP的新生血管增生阶段,应用抗VEGF类药物抑制VEGF可以阻断视网膜血管的异常增生.本文就抗VEGF类药物贝伐单抗(Avastin)单纯玻璃体内注射、玻璃体内注射联合视网膜光凝或玻璃体切除手术等不同方式治疗ROP的临床研究进行回顾,并就贝伐单抗治疗ROP的安全性评价进行汇总.