D-Timolol和L-Timolol对大鼠脉络膜新生血管及内皮细胞的作用(英文)

目的:老年黄斑变性患者存在脉络膜血流灌注障碍。据此,我们推测血管活性药物,由于能减小脉络膜血流的阻力,可能会防止脉络膜新生血管的发展。D-Timolol和L-timolol是应用于心血管和青光眼治疗的降血压药物。本文旨在评价二者对激光诱发的大鼠脉络膜新生血管模型和人脐静脉内皮细胞(HUVEC)的作用。方法:雄性BrownNorway大鼠,麻醉下行Nd:YAG激光击穿Bruch膜。激光后,予D-Timolol和L-Tim-olol每日一次腹腔注射4wk。2wk末及4wk末时行荧光造影检查。观察不同浓度D-Timolol和L-Timolol对体外培养的HUVEC细胞增殖和黏附的影响。结果:在激光诱发的大鼠脉络膜新生血管模型中,予D-Timolol腹腔注射15mg/(Kg·d),可减少荧光渗漏的位点,至对照组的83%。而L-Timolol对脉络膜新生血管的形成没有影响,即使注射更高的治疗剂量20mg/(kg·d)。D-Timolol300mg/L可抑制内皮细胞的生长。L-Timolol在1000mg/L可以显著抑制细胞的增殖,但在相对低的浓度300mg/L,则没有发现明显的抑制作用。在HUVEC细胞黏附的观察中,两者均未有显著的影响。结论:D-Timolol可减少激光诱发的脉络膜新生血管的形成,也能抑制血管内皮细胞的增殖。而L-Tim-olol在同样的浓度不影响内皮细胞的增殖,也不能影响大鼠脉络膜新生血管的形成。这两种同分异构体对动物模型和培养细胞的不同作用,可能对探索脉络膜新生血管的治疗有新的意义。     

Z,E-butlidedephthalide对大鼠脉络膜新生血管和兔眼血流的影响

目的:研究Z,E-butylidedephthalide(Bdph)对激光诱导的大鼠脉络膜新生血管(CNV)和兔眼脉络膜血流的影响.方法:雄性Brown Norway大鼠行Nd:YAG激光诱导眼底Bruch膜破裂,而后分别予30 mg/kg和15 mg/kg Bdph 1 次/d腹腔注射,连续4 wk.2 wk及4 wk末行眼底荧光血管造影检查.4 wk末制作脉络膜平片测量新生血管的面积.用1 g/L Z,E-butylidenephthalide给雌性新西兰大白兔点眼,采用彩色微球技术测量大白兔眼部血流的变化.结果:和对照组相比,Bdph 30 mg/kg和15 mg/kg组大鼠眼底血管荧光渗漏明显减少,P＜0.01;在此两组中,眼底荧光血管造影测定的新生血管面积以及脉络膜平片测定的新生血管面积都比对照组减少.10 g/L Z,E-butylideneph-thalide滴入兔眼30和60 min后脉络膜血流均比对照组增加,P＜0.05.结论:Z,E-butylidedephthalide可以抑制大鼠脉络膜新生血管的生长,增强兔眼脉络膜血流,可能成为治疗年龄相关性黄斑变性的新药.     

柚皮素对大鼠眼血流和脉络膜新生血管的作用(英文)

目的:研究柚皮素对激光诱发大鼠脉络膜新生血管形成(CNV)、高眼压兔眼血流和缺血大鼠眼视网膜功能恢复的作用。方法:选择雄性棕色挪威大鼠,采用激光诱发Bruch膜破裂后,予10g/L(20mg/kg)柚皮素,1次/d,持续4wk;光凝后2,4wk分别做眼底荧光血管造影,评估CNV的形成。采用彩色微球技术、眼电生理技术检测兔眼血流和大鼠视网膜功能恢复。结果:与对照组比较,10g/L柚皮素能明显增加高眼压兔眼脉络膜血流(P<0.05),能明显增加缺血大鼠眼视网膜功能恢复(P<0.05),能明显减轻光凝点荧光素渗漏(75.8%～95.0%,P<0.01)。结论:柚皮素能抑制激光诱发大鼠络膜新生血管形成;增加高眼压兔眼脉络膜血流;增加缺血大鼠眼视网膜功能恢复。     

氪激光诱发大鼠脉络膜新生血管的基因表达谱

目的研究氪激光诱发大鼠脉络膜新生血管的基因表达谱。方法对氪激光诱发BN大鼠脉络膜新生血管的脉络膜组织和正常BN大鼠的脉络膜组织进行总RNA的提取,将纯化后的mRNA进行逆转录制备杂交探针,应用含有5705条Oligo DNA的表达谱芯片对氪激光诱发BN大鼠脉络膜新生血管的脉络膜组织和正常BN大鼠的脉络膜组织进行差异表达谱分析。结果在氪激光诱发BN大鼠脉络膜新生血管的基因表达谱中差异表达基因共有46条,其中上调的基因3条,下调的基因43条。它们分别是物质转运、信号传导、代谢、发育、分化、细胞黏附相关基因。结论氪激光诱发脉络膜新生血管的发生发展涉及多基因改变。（中华眼科杂志,2005,41：317-320）     

胼酞嗪对眼血流和激光诱导的脉络膜新生血管及体外培养的内皮细胞的影响

目的：研究胼酞嗪对兔脉络膜血流和激光诱导的鼠脉络膜新生血管及人脐静脉内皮细胞管状结构形成的影响。方法：雌性新西兰白兔左眼眼内压升高至40mmHg后，滴入10g／L胼酞嗪滴眼液，采用彩色微球技术测量眼血流变化。用Nd：YAG激光诱导雄性BrownNorway大鼠至Bruch膜破裂，而后分别予生理盐水或5，10和20g/L胼酞嗪滴眼液点眼，3次／d，连续4wk。采用眼底荧光血管造影和脉络膜平片测量新生血管的面积。此外亦用不同浓度的胼酞嗪作用于培养之人脐静脉内皮细胞并探讨对其管状结构形成的影响。结果：10g／L胼酞嗪滴眼液滴入眼内压为40mmHg的兔眼30，60min后，脉络膜血流明显增加。眼底荧光血管造影和脉络膜平片测量均显示，经过4wk药物治疗，5，10，20g／L胼酞嗪滴眼液均明显抑制了鼠眼CNV的形成。3—30mg／L胼酞嗪对在基质凝胶中培养了48h的人脐静脉内皮细胞的管状结构的形成有抑制作用。结论：胼酞嗪可以抑制体内脉络膜新生血管及体外培养的人脐静脉内皮细胞管状结构的形成，并增强缺血后的兔脉络膜血流。胼酞嗪有望成为治疗年龄相关性黄斑变性的药物。     

ZA-76对大鼠脉络膜新生血管形成、兔眼血流及血管内皮细胞的影响

目的观察ZA-76对激光诱发大鼠脉络膜新生血管(choroidal neovascularization,CNV)形成、高眼压兔眼血流及人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)增殖的影响。方法氪激光诱发雄性棕色挪威大鼠CNV。30只(60眼)模型大鼠随机分为治疗组和对照组,每组15只(30眼)。治疗组在光凝后第2天开始腹腔注射20g·L-1ZA-76(20mg·kg-1),对照组给予等体积的溶媒二甲基亚砜,每天1次,时间均为4周。光凝后2周、4周分别行眼底照相和眼底荧光血管造影,评估CNV的形成率。光凝后4周处死大鼠,取材作病理切片,用于HE染色及免疫组织化学检测VEGF、COX-2的表达;用彩色微球技术检测20g·L-1ZA-76(50μL)对高眼压兔眼血流的影响;MTT法检测不同浓度ZA-76(100mg·L-1、200mg·L-1、400mg·L-1)对HUVEC细胞增殖的影响。结果治疗组光凝后2周、4周CNV形成率分别为80.83%、79.58%,明显低于对照组的94.58%、95.00%(均为P<0.01);治疗组VEGF、COX-2阳性染色密度分别为(51.13±6.15)mm-2、(37.22±5·18)mm-2,低于对照组[分别为(71.55±9.89)mm-2、(58.78±7.02)mm-2(均为P<0.01)];治疗组脉络膜血流在30min、60min、120min均明显高于对照组;不同浓度ZA-76干预48h,各浓度组ZA-76对HUVEC细胞增殖抑制作用均强于对照组,以400mg·L-1ZA-76作用更显著。结论 ZA-76能抑制激光诱导大鼠CNV形成,抑制VEGF、COX-2的表达;抑制血管内皮细胞增生;改善脉络膜血流。     

丙烯醛诱导大鼠脉络膜新生血管的形成

目的：：探讨丙烯醛( AC)诱导大鼠脉络膜新生血管( CNV)的形成。方法：选取远交群大鼠( SD)12只,随机将大鼠分为三组,即空白组,AC诱导4wk组和AC诱导8wk组,其中空白对照组每日200μL新鲜自来水灌胃,实验组为200μL AC溶液(2.5 mg/kg/d)每日灌胃,分为诱导4wk组和8wk组,取材后视网膜组织石蜡包埋,切片及HE染色。结果：空白对照组及AC诱导4 wk组均见RPE-Bruch膜连续性完整,未见明显异常;AC 诱导8 wk 组发现 RPE-Bruch膜连续性缺失,脉络膜新生血管长入视网膜神经上皮层内。结论：长期使用AC可以诱导大鼠脉络膜新生血管形成。     

柚皮铜配合物对激光诱发大鼠脉络膜新生血管形成的抑制作用

目的：观察柚皮铜配合物对激光诱发大鼠脉络膜新生血管形成的抑制作用。

方法：选取雄性Brown-Norway大鼠24只,其中18只建立大鼠脉络膜新生血管模型,6只12眼作正常对照。动物模型随机分为模型对照组和2个不同给药实验组,每组6只。模型建立后,实验组大鼠腹腔注射同等剂量的柚皮素、柚皮铜配合物\〖20mg/(kg·d)\〗,对照组不给药。于造模后30d取各组视网膜-脉络膜-巩膜复合体标本,在荧光显微镜下观察脉络膜新生血管的面积,采集各组大鼠的脉络膜标本,采用RT-PCR及Western blot方法检测各组大鼠脉络膜组织中VEGF、COX-2、PI3K、MAPK、MMP-2、MMP-9mRNA与蛋白的表达。

结果：与模型对照组相比,各实验组大鼠新生血管面积均减少,但柚皮铜配合物组较柚皮素组更明显,差异均具有显著统计学意义(P<0.01); 模型组基因和蛋白表达水平均高于正常对照组,差异均有统计学意义(P<0.01); 各实验组基因及蛋白表达水平与模型组相比均有所下降,除PI3K,MMP-9外,柚皮铜配合物组下降幅度大于柚皮素组,差异均有统计学意义(P<0.05)。

结论：相较于柚皮素,柚皮铜配合物有更好的生物活性,对激光诱发大鼠脉络膜新生血管的形成有更显著的抑制作用。     

Tetramethylpyrazine在视网膜色素上皮细胞变性及脉络膜血流和RPE细胞氧化应激中的作用

目的:研究1%Tetramethylpyrazine(TMP)在视网膜色素上皮细胞(RPE)变性,脉络膜血流和RPE细胞氧化应激中的作用。方法:在碘酸钠诱导的大鼠RPE变性研究中,1%TMP滴眼液预先处理1wk,3次/d,1wk后予碘酸钠舌下静脉注射,在2wk和4wk末,视网膜电图(ERG)测量c波。色素微球体技术分析高眼压状态下TMP对脉络膜血流的影响。Methylthiazoltetrazolium(MTT)分析TMP在各种氧化应激中对RPE的保护作用。结果:碘酸钠注射后2wk,碘酸钠组ERG的c波下降至对照组的36%(P<0.01)。4wk后,碘酸钠组下降至对照组的46%(P<0.01),而1%TMP+碘酸钠组下降至对照组的77%(P<0.01)。与碘酸钠组比较,1%TMP+碘酸钠组控制了67%的c波下降(P<0.05)。在脉络膜血流的测量中,30,60,和120min的结果显示,TMP显著增加脉络膜血流。在氧化应激部分,不同浓度的TMP在各种氧化应激损伤中,对RPE都有各种程度的保护作用。结论:浓度为1%Tetramethylpyrazine可以显著保护碘酸钠和氧化应激诱导的RPE变性,增加脉络膜血流,并可能在AMD的治疗中发挥作用。     

Tetramethylpyrazine在视网膜色素上皮细胞变性及脉络膜血流和RPE细胞氧化应激中的作用

目的：研究1% Tetramethylpyrazine （TMP）在视网膜色素上皮细胞（RPE）变性,脉络膜血流和RPE细胞氧化应激中的作用.方法：在碘酸钠诱导的大鼠RPE变性研究中,1%TMP滴眼液预先处理1wk,3次/d,1wk后予碘酸钠舌下静脉注射,在2wk和4wk末,视网膜电图（ERG）测量c波.色素微球体技术分析高眼压状态下TMP对脉络膜血流的影响.Methylthiazoltetrazolium （MTT）分析TMP在各种氧化应激中对RPE的保护作用.结果：碘酸钠注射后2wk,碘酸钠组ERG的c波下降至对照组的36%（P〈0.01）.4wk后,碘酸钠组下降至对照组的46%（P〈0.01）,而1% TMP ＋碘酸钠组下降至对照组的77%（P〈0.01）.与碘酸钠组比较,1%TMP ＋碘酸钠组控制了67%的c波下降（P〈0.05）.在脉络膜血流的测量中,30,60,和120min的结果显示,TMP显著增加脉络膜血流.在氧化应激部分,不同浓度的TMP在各种氧化应激损伤中,对RPE都有各种程度的保护作用.结论：浓度为1% Tetramethylpyrazine可以显著保护碘酸钠和氧化应激诱导的RPE变性,增加脉络膜血流,并可能在AMD的治疗中发挥作用.     

Effectof d-tiｍolol and l-tiｍololon rat experiｍental choroidal neovasculariｚation in vivo and endothelial cells in vitro

ImPairmentof choroidal Perfusion was found in age-related macular degeneration(AMD) Patients. We Postulatedthat vasoactive agents, which can reduce choroidal blood flow resistance, might Preventthe develoPmentof choroidal neovascularization (CNV). D-Timolol and L-Timolol are hyPotensive agents used in cardiovascular and glaucomatheraPy.their effectson laser-induced exPerimental CNV rat model and human umbilical vein endothelial cells (HUVEC) werethus evaluated. ·METHODS: Male Brown Norway rats were anesthetizedto receive Nd:YAG laserto breakthe Bruch's membrane. D-Timolol and L-Timolol were givenonce dailythrough intraPeritoneal injection after lasertreatment for 4 weeks. Fluorescein angiograPhy(FA) was Performedon 2 weeks and 4 weeks. HUVEC weretested by Proliferation assay and adhesion assay with D-Timolol and L-Timolol at different concentrations. ·RESULTS: D-Timolol reducedthe fluorescein leakageto 83%ofthe control grouP in laser-induced rat's CNV model at a dosageof 15mg/(kg·d). L-Timolol had no effecton CNV formation even at a higher dosageof 20mg/(kg·d). D-Timolol inhibitedthe endothelial cells Proliferation significantly by 300mg/L. L-Timolol also significantly inhibitedthe cell Proliferation at 1 000mg/L. But at a lower dose such as 300mg/L, no significant inhibitory effect was found. Both drugs showed no effecton cell adhesion function in cell culture exPeriments. ·CONCLUSION: D-Timolol was foundto Prevent CNV develoPment in laser-induced model in vivo and inhibit vascular endothelial cells Proliferation in vitro . L-Timolol had no effecton cell Proliferation atthe same dose, and neitheron rat CNV model.the results indicatethesetwo isomers have different functionson rat's CNV Prevention andon HUVEC cell Proliferation.     

Apigenin inhibits laser-induced choroidal neovascularization and regulates endothelial cell function.

PURPOSE: The aim of this study is to evaluate the effect of apigenin on laser-induced experimental choroidal neovascularization (CNV) rat model and endothelial cell proliferation and migration. METHODS: Male Brown Norway rats were anesthetized to receive Nd:YAG laser to break the Bruch's membrane. Apigenin at 5, 15, or 30 mg/kg was given once-daily through intraperitoneal injection after laser treatment for 4 weeks. The development of CNV was determined by fluorescein angiography performed on weeks 2 and 4. Endothelial cell function was evaluated with proliferation assay and migration assay. RESULTS: The intensity of fluorescein leakage from the photocoagulated lesions decreased significantly, compared to the control group, following apigenin treatment. Four (4) weeks after administration, apigenin, at 15 and 30 mg/kg, inhibited CNV development to 84.5% and 83.6% of the control group, respectively (P<0.05). Apigenin also interfered with the endothelial cells' proliferation and migration. At 3 and 10 microg/ml, apigenin inhibited the growth of human umbilical vein endothelial cells (HUVEC) to 54% and 46% of the control group, respectively (P<0.01); inhibited the growth of choroidal endothelial cells to 47% and 8% of the control group, respectively (P<0.01); and inhibited HUVEC migration over 50%, compared with the control (P<0.01). CONCLUSIONS: Apigenin exerts an inhibitory effect on choroidal angiogenesis in vitro and in vivo. It should be further evaluated for its potential as a novel therapy for CNV.     

Effects of hydralazine on ocular blood flow and laser-induced choroidal neovascularization

AIM: To investigate the effect of hydralazine on choroidal blood flow in rabbits and laser-induced choroidal neovascularization (CNV) in rats and on tube formation of human umbilical vein endothelial cells (HUVEC). METHODS: Female New Zealand white rabbits were used with raised intraocular pressure (IOP) of the left eye to 40mmHg. Hydralazine (10g/L) eye drops were instilled and ocular blood flow was measured with colored microspheres technique. Male Brown Norway rats were treated with Nd:YAG laser to break Bruch's membrane. Hydralazine (5, 10, 20g/L) eye drops or saline alone was instilled three times a day for 4 weeks after laser treatment. Fluorescein angiography (FA) and choroidal flat mount were used to measure the area of CNV. Tube formation of HUVEC was studied at different concentrations of hydralazine. RESULTS: With raised IOP to 40mmHg on rabbits, 10g/L hydralazine eye drops enhanced the choroidal blood flow significantly at 30 and 60 minutes after drug instillation. After 4 weeks of drug treatment, 5, 10 and 20g/L hydralazine eye drops all reduced the CNV formation dramatically measured by fluorescein angiography and choroidal flat mount. When HUVEC was cultured on matrix gel for 48 hours, the tube formation of HUVEC were prevented.by hydralazine at 3-30mg /L. CONCLUSION: Hydralazine prevents CNV formation in vivo and HUVEC tube formation in vitro, and enhances rabbits' choroidal blood flow after ischemia. It is hoped that hydralazine could be used to treat age-related macular degeneration in the future.     

Effect of interleukin-1 blockers, CK112, and CK116 on rat experimental choroidal neovascularization in vivo and endothelial cell cultures in vitro.

PURPOSE: The aim of this study was to investigate the effect of interleukin-1 blockers, CK112 and CK116, on laser-induced experimental choroidal neovascularization (CNV) in rat models in vivo and endothelial cell proliferation in vitro. METHODS: Male Brown Norway rats were anesthetized to receive Nd:YAG laser to break the Bruch's membrane. CK112, CK116, and prednisolone were given once-daily through intraperitoneal (i.p.) injection after laser treatment for 4 weeks. The development of CNV was determined by fluorescein angiography performed on weeks 2 and 4. Human umbilical vein endothelial cells (HUVEC) were tested with proliferation assay with CK112, CK116, and prednisolone at different concentrations. RESULTS: The intensity of fluorescein leakage from the photocoagulated lesions decreased significantly, compared to the control group (treated with dimethyl sulfoxide [DMSO] only), following CK112, CK116, and prednisolone treatment. Four (4) weeks after administration, CK112, at 10 mg/kg and 30 mg/kg, inhibited CNV development to 75% and 77% of the control group, respectively (P < 0.01). Both CK116, 10 mg/kg, and prednisolone, 5 mg/kg, inhibited the CNV development to 85% of the control group (P < 0.05). All three compounds interfered with the endothelial cell proliferation significantly. The reduction of the endothelial cells was 50.5% (P < 0.01), 28.5% (P < 0.05), and 23.1% (P < 0.05), respectively, in 500 microg/mL, 300 microg/mL, and 100 microg/mL of the CK112-treated group. CK116 inhibited the cell proliferation significantly to 77.2% of the control group at 500 microg/mL (P < 0.05). CONCLUSIONS: CK112 and CK116 inhibited the development of CNV in the rat model and interfered with vascular endothelial cell proliferation in vitro. Our results suggest that CK112 and CK116 may be good candidates to inhibit ocular neovascularization related to age-related macular degeneration (ARMD).     

Effect of tetramethylpyrazine on rat experimental choroidal neovascularization in vivo and endothelial cell cultures in vitro

PURPOSE: To evaluate the effect of tetramethylpyrazine (TMP) on laser-induced experimental choroidal neovascularization (CNV) in rat model in vivo and on endothelial cell proliferation in vitro. METHODS: Male Brown Norway rats were anesthetized to receive Nd:YAG laser to break the Bruch membrane. TMP was given once daily through intraperitoneal injection after laser treatment for 4 weeks. The development of CNV was determined by angiography performed on week 2 and week 4 using sodium fluorescein (FA) or fluorescein isothiocyanate-dextran (FD70-FA). Human umbilical vein endothelial cells (HUVECs) were tested with proliferation assay with TMP at different concentrations. RESULTS: According to the angiograms of FA, intensity of fluorescein leakage from the photocoagulated lesions decreased significantly after TMP treatment. The number of rats with less leaky points (相似文献   

Inhibition of choroidal neovascularization by blocking vascular endothelial growth factor receptor tyrosine kinase

Purpose To investigate the role played by receptors of vascular endothelial growth factors, Flt-1 and KDR/Flk-1, on an experimental model of choroidal neovascularization (CNV). Methods The vascular endothelial growth factor-A (VEGF-A) receptor-specific tyrosine kinase inhibitor SU5416 was administered to a laser-induced mouse model of CNV. The formation of CNV and the degree of vascular permeability in Flt-1 tyrosine kinase domain-deficient mice were also investigated. Results SU5416 reduced vascularity and vascular endothelial cell proliferation, and promoted endothelial cell apoptosis within CNV. Furthermore, the formation of CNV and the degree of vascular permeability were significantly reduced in Flt-1 tyrosine kinase domain-deficient mice, and this effect was enhanced by the administration of SU5416. Conclusions Both Flt-1 and KDR/Flk-1 have a significant role in CNV formation. Suppression of apoptosis may be involved in the process.     

Soluble EphB4 regulates choroidal endothelial cell function and inhibits laser-induced choroidal neovascularization

PURPOSE: The purpose of this study was to evaluate the effect of a soluble monomeric form of the EphB4 extracellular domain (sEphB4) on choroidal endothelial cell (CEC) migration and tube formation and on experimental laser-induced choroidal neovascularization (CNV). METHODS: EphrinB2 and EphB4 expression in CECs was investigated by Western blot analysis and immunohistochemistry. Effects of sEphB4 (0.5-3 microg/mL) on CEC migration were evaluated with a modified Boyden chamber assay. Tube formation was assayed in CEC cultures in collagen gel. CNV was induced in rats by laser photocoagulation. The effects of intravitreal injection of sEphB4 on CNV development were evaluated at day 14 by fluorescein angiography (FA), confocal volumetric analysis of isolectin-B4 labeled flatmounts, and histologic examination of CNV membranes. RESULTS: CEC cells express both EphB4 and EphrinB2, according to Western blot analysis. Immunohistochemical sections of rat eye showed immunoreactivity for both EphB4 and EphrinB2 in the choroidal endothelium. sEphB4 reduced CEC migration in response to vascular endothelial growth factor (P < 0.01). Similarly, sEphB4 inhibited CEC tube formation in a dose-dependent manner. EphB4, and to a lesser extent EphrinB2, were detected on vascular channels within laser-induced CNV membranes. Intravitreal injection of sEphB4 inhibited laser-induced CNV formation. CNV membranes showed a reduction in leakage score (P < 0.05), and membrane volumes were reduced in size (P < 0.05). Histologic analysis revealed that vascularity was reduced in sEphB4-treated membranes. CONCLUSIONS: Recombinant soluble monomeric EphB4 exerts an inhibitory effect on choroidal angiogenesis in vitro and in vivo. It should be further evaluated for its potential as a novel therapy for CNV.     

脉络膜新生血管动物模型的建立与评估

目的:评价氪激光诱导棕色挪威(brown norway,BN)大鼠脉络膜新生血管(choroidal neovascularization,CNV)模型的可行性。方法:BN大鼠32只一眼行氪激光(659nm)眼底光凝,另一眼作空白对照。激光功率、光凝斑直径和曝光时间分别为360mW、50μm及0.05s。分别在光凝后7,14,21,28d随机选取7只大鼠行眼底照像、荧光素眼底血管造影(FFA)检查。随机选取1只大鼠行脉络膜血管铺片检查。眼球标本行组织病理切片光镜、免疫组织化学染色观察。结果:光凝后7d,CNV开始形成,21d达到高峰,14~28dCNV无明显变化。7~21d,FFA显示造影晚期荧光素渗漏及光镜下CNV厚度逐渐增加,28d时可能因瘢痕开始形成,故FFA渗漏和CNV厚度开始减少。结论:氪激光诱导BN大鼠的CNV模型是一种较为理想的模型,FFA及CNV厚度分析是CNV定量研究的有效手段。     

Anti-angiogenic effects of non-peptide integrin αvβ3 specific antagonist on laser-induced choroidal neovascularization in mice

Background  To evaluate the anti-angiogenic effects of integrin αvβ3 specific antagonist BS-1417 on laser-induced choroidal neovascularization (CNV) in mice. Methods  Male C57BL/6 mice were treated with daily intraperitoneal injections of BS-1417 or saline starting at the onset (day 0) of experiments. CNV was induced by laser photocoagulation the next day. Fluorescein angiograms (FA) and choroidal flatmount FITC-dextran perfusion were performed on experimental day 8. Histological and immunohistochemical examinations were performed with consecutive cryosections. Sub-confluent human vascular endothelial cells (HUVEC) were grown in vitro under various concentrations (0–10 μg/ml) of BS-1417 and the numbers of cell were measured at 48 hours of incubation. After fixation, immunocytochemistry was performed. Results  BS-1417 significantly decreased the area of dye leakage determined by FA (40% of control, p = 0.0008) and reduced the CNV size found on choroidal flatmount (18% of control, p = 0.007). In histological findings, BS-1417 apparently suppressed the size of laser-induced CNV. Immunoreactivities for VEGF and integrin αv were remarkably attenuated with BS-1417 compared to control. BS-1417 inhibited the growth and VEGF expression of HUVEC in vitro. Conclusions  The integrin αvβ3 may play a key role in the induction of laser-induced CNV. The antagonists for integrin αvβ3 may have therapeutic effects in CNV associated diseases such as age-related macular degenerations.