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。     

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。     

D-Timolol和L-Timolol对大鼠脉络膜新生血管及内皮细胞的作用

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Brown Norway大鼠,麻醉下行Nd:YAG激光击穿Bruch膜.激光后,予D-Timo lo l和L-Timolol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1 5mg/(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-Timolol 300mg/L可抑制内皮细胞的生长.L-Timolol在1 000mg/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-Timolol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.     

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。     

实验性脉络膜新生血管的定量评价

目的 评价氪激光诱导下大鼠脉络膜新生血管(choroidal neovascularization,CNV)面积计算的可行性.方法 采用氪激光击破大鼠右眼 Bruch 膜的方法诱导24只大鼠产生实验性 CNV,另一眼作正常对照眼.眼底彩照及眼底血管荧光造影检查(fundus fluorescein angiography,FFA)分别于光凝后1周、2周、3周、4周时观察大鼠眼底CNV渗漏情况,脉络膜铺片技术测量CNV面积.结果 氪激光大鼠CNV模型成模率3周、4周时稳定(75.00%、78.75%).从FFA、脉络膜血管铺片技术可见大鼠CNV面积随造模后时间延长呈明显递增生长(P＜0.05),3周、4周时差异不明显,病理切片亦可见CNV中新生血管数量的增多.结论 FFA检查、脉络膜血管铺片技术、病理切片均可作为评价CNV生长的指标,其中脉络膜血管铺片技术是CNV定量评价的可靠方法.     

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。     

新型光敏剂酞菁锌在兔眼组织分布的研究

目的探讨新型光敏剂酞菁锌在色素兔脉络膜和视网膜分布情况及其浓度的高峰期,为用金属酞菁介导的光动力疗法(PDT)提供相关的参考方案。设计实验研究。研究对象健康有色家兔28只。方法6只兔静脉注射不同剂量酞菁锌药后进行眼底血管造影观察。对照组(2只)经耳缘静脉注入等量生理盐水,实验组(20只)经耳缘静脉注入酞菁锌20mg/kg,在给药后5、10、15、30和60min摘除眼球作冰冻切片(每组8眼),荧光显微镜下观察眼球的荧光分布。主要指标眼底血管造影荧光充盈情况及冰冻切片荧光程度。结果注射20mg/kg酞菁锌后10～20s,荧光造影显示脉络膜渐呈弥漫高荧光,随时间推移荧光强度逐渐减弱。荧光显微镜下观察给药后10～20min为酞菁锌在眼组织分布的高峰期,30min后药物荧光有所下降,视网膜层仍有少量荧光,60min药物荧光基本已消退。结论酞菁锌主要迅速分布于脉络膜和视网膜色素上皮层,而其他组织如内层视网膜、巩膜等药物荧光分布少。提示酞菁锌介导的PDT治疗脉络膜新生血管最佳时间是给药后10～20min进行光照。     

小鼠的脉络膜新生血管模型

目的建立激光诱导的小鼠的脉络膜新生血管模型,观察其自然演变过程.方法通过氩激光光凝诱导C57BL/6J小鼠产生脉络膜新生血管,光凝后1 wk、2wk、4wk、2 mo、3 mo、4mo、5mo、6 mo行荧光素眼底血管造影检查,并且在光凝后1 wk、2wk、4wk、2mo、3 mo、6 mo处死小鼠(n=2),摘除眼球,行组织学检查.结果光凝后1 wk、2 wk、4 wk、2 mo、3 mo、4mo、5mo、6 mo荧光素眼底血管造影显示有荧光渗漏的光凝斑的百分率分别为59.0%(46/78)、62.1%(41/66)、44.4%(24/54)、42.9%(18/42)、46.7%(14/30)、44.5%(8/18)、38.9%(7/18)、22.2%(4/18),组织学检查显示光凝斑内有新生血管形成,随着时间的延长,视网膜色素上皮细胞呈不同程度增生,可完全包绕脉络膜新生血管(choroidal neovascularization,CNV).结论氩激光光凝可成功诱导小鼠的CNV模型,CNV在光凝后2wk达到高峰.     

Naringenin在碘酸钠诱导的视网膜色素上皮细胞变性和激光诱导的脉络膜新生血管鼠模型中的作用

目的：研究naringenin滴眼对碘酸钠诱导的大鼠视网模色素上皮（retinal pigment epithelium,RPE）变性以及对激光诱导的脉络膜新生血管（choroidal neovascularization,CNV）的作用。方法：10g/L naringenin滴眼液预先处理1wk（3次／d）,1wk后予35mg／kg碘酸钠舌下静脉注射诱导大鼠RPE变性,在2wk和4wk末,视网膜电图（electroretinogram,ERG）测量C波。另预处理1wk（3次／d）,在2wk和4wk末用荧光素血管造影（fluoreseein angiography,FA）和荧光显微镜测量CNV面积。结果：碘酸钠注射后2wk,碘酸钠组ERG的C波下降至对照组的53％（P〈0．01）。而naringenin＋碘酸钠组则无明显变化。4wk后,碘酸钠组下降至对照组的37％（P〈0．01）,而naringenin＋碘酸钠组下降至对照组的57％（P〈0．01）。与碘酸钠组比较,naringenin＋碘酸钠组控制了66％的C波下降（P〈0．05）。35mg／kg naringenin组FA测量的CNV面积2,4wk末分别是对照组的53％和49％（P〈0．01）。4wk后naringenin组荧光显微镜测量的CNV面积是对照组的47％（P〈0．01）。结论：10g/L naringenin可以显著保护碘酸钠诱导的RPE变性,也能减小CNV的形成。     

Naringenin在碘酸钠诱导的视网膜色素上皮细胞变性和激光诱导的脉络膜新生血管鼠模型中的作用

目的:研究naringenin滴眼对碘酸钠诱导的大鼠视网膜色素上皮(retinal pigment epithelium,RPE)变性以及对激光诱导的脉络膜新生血管(choroidal neovascularization,CNV)的作用。方法:10g/Lnaringenin滴眼液预先处理1wk(3次/d),1wk后予35mg/kg碘酸钠舌下静脉注射诱导大鼠RPE变性,在2wk和4wk末,视网膜电图(electroretinogram,ERG)测量C波。另预处理1wk(3次/d),在2wk和4wk末用荧光素血管造影(fluorescein angiography,FA)和荧光显微镜测量CNV面积。结果:碘酸钠注射后2wk,碘酸钠组ERG的C波下降至对照组的53%(P<0.01)。而naringenin+碘酸钠组则无明显变化。4wk后,碘酸钠组下降至对照组的37%(P<0.01),而naringenin+碘酸钠组下降至对照组的57%(P<0.01)。与碘酸钠组比较,naringenin+碘酸钠组控制了66%的C波下降(P<0.05)。35mg/kgnaringenin组FA测量的CNV面积2,4wk末分别是对照组的53%和49%(P<0.01)。4wk后naringenin组荧光显微镜测量的CNV面积是对照组的47%(P<0.01)。结论:10g/Lnaringenin可以显著保护碘酸钠诱导的RPE变性,也能减小CNV的形成。     

Effect of Z,E-butylidedephthalide on experimental choroidal neovascularization in rat and ocular blood flow in rabbits

To investigate the effect of Z,E-butylidedephthalide (Bdph)on laser-induced experimental choroidal neovasculari- zation (CNV) in rat model and choroid blood flow in rabbits' eyes. · METHODS: Male Brown Norway rats were treated with Nd:YAG laser to break Bruch's membrane. 30mg/kg and 15mg/kg Bdph were given daily through intraperitoneal injection for 4 weeks after laser treatment. Fluorescein angiography (FA) and choroidal flat mount were used to measure the development of CNV. Female New Zealand white rabbits' eyes were instilled with 10g/L Z,E-BdPh solution, and ocular blood flow was measured with colored microsphere technique. · RESULTS: The intensity of fluorescein leakage, indicating the ocular lesion, decreased significantly in group Bdph 30mg/kg and 15mg/kg, as compared to the control at P <0.01. The area of neovascularization checked by FA in both groups of Bdph, at 30mg/kg and 15mg/kg decreased significantly compared to the control group at P <0.05. On the choroid flat mount, the areas of CNV were also smaller in both Bdph groups than that in control group. One percent drug solution instilled into rabbits' eyes could improve the choroid blood flow at 30 and 60 minutes after drug instillation (P <0.05). · CONCLUSION: Z,E-butylidedephthalide can inhibit the development of CNV in the rat eyes and increase the choroid blood flow in the rabbit eyes. These results suggest that Z,E-butylidedephthalide may be a good agent for the treatment of age-related macular degeneration (AMD).     

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.     

黄酮对眼部血流量和脉络膜新生血管的作用

目的:观察黄酮对兔眼部血流量和大鼠脉络膜新生血管(CNV)的作用。方法:在体研究中,我们采用彩色微球技术测定黄酮对兔眼部血流量的影响。预先给予5g/L黄酮滴眼液(3次/d),1wk后用激光诱导大鼠CNV的生成,在2和4wk末,采用荧光血管造影(FA)测量CNV面积。离体研究中,采用MTT法检测黄酮对人脐静脉内皮细胞(HUVECs)存活率的影响。结果:黄酮显著的增加了兔眼部血流量,抑制了由激光诱导的CNV的生成。离体研究结果表明,黄酮抑制了HU-VEC的增生。结论:黄酮能够增加眼部血流量,并且抑制CNV的生成。     

Effect of flavone on the ocular blood flow and formation of choroidal neovascularization

AIM: To investigate the effect of flavone on ocular blood flow in rabbit eyes and the formation of choroidal neovascularization (CNV) in rat model of age-related macular degeneration (AMD). METHODS: In in vivo studies, colored microsphere technique was used to determine the ocular blood flow in ocular hypertensive rabbit eyes. The rat eyes were treated with 0.5% flavone eye drops 3 times a day for 1 week before and 4 weeks after laser-induced injury of Bruch's membrane. The development of CNV was determined by fluorescein angiography (FA) performed on the 2nd and 4th after injury. In in vitro studies, the effect of flavone on the viability of human umbilical vein endothelial cells HUVECs was measured by MTT assay. RESULTS: The ocular blood flow in rabbit eyes was significantly increased after flavone instillation. Flavone significantly inhibited the formation of laser induced CNV. In vitro results showed that flavone inhibited the proliferation of HUVECs. CONCLUSION: Flavone could increase ocular blood flow and inhibit the formation of CNV.     

Effects of naringenin on ocular blood flow and choroidal neovascularization in experimental animals

AIM: To investigate the effects of naringenin on laser- induced experimental choroidal neovascularization (CNV) in rat models, ocular blood flow and retinal function recovery after ischemic insults in rat eyes. METHODS: Male Brown Norway rats were treated to break the Bruch's membrane. Naringenin 10g/L (20mg/kg) was given once per day through intraperitoneal injection for 4 weeks after laser treatment. The development of CNV was determined by fluorescein angiography (FA) performed on week 2 and 4. The colored microsphere technique and electroretinography method were used for the study of ocular blood flow and retinal function recovery, respectively. RESULTS: The choroidal blood flow in elevated intraocular pressure (IOP) rabbit eyes was significantly increased by 10g/L naringenin solution as compared to control group (P < 0.05). The retinal function recovery after ischemic insults in rat eyes indicated significant increase of b-wave recovery in treated group, as compared to control group (P <0.05). The intensity of fluorescein leakage from the photocoagulated lesions significantly decreased in treated group, compared to the control group (75.8%-95.0%, P <0.01). CONCLUSION: Naringenin could prevent the development of CNV on laser-induced experimental rat models, increase the choroidal blood flow in elevated IOP rabbit eyes and be beneficial on retinal function recovery in ischemic rat eyes.     

Effect of naringenin on NaIO3-induced retinal pigment epithelium degeneration and laser-induced choroidal neovascularization in rats

AIM: To study the effects of naringenin eye drops on NaIO3-induced retinal pigment epithelium (RPE) degeneration and laser-induced choroidal neovascularization (CNV) in rat eyes. · METHODS: The 35mg/kg NaIO3-induced RPE degeneration was prevented by 10g/L naringenin eye drops 3 times a day for 7 days in advance of NaIO3 injection, and then 2 to 4 weeks thereafter, RPE function was measured with C-wave of electroretinogram (ERG). The laser-induced CNV rats were treated with laser to break the Bruch's membrane and the CNV formation was prevented by 10g/L naringenin eye drops instilled 3 times a day for 2 to 4 weeks. The CNV formation was measured with fluorescein angiography (FA) and flat mount. · RESULTS: Two weeks after NaIO3 injection, the amplitude of ERG C-wave fell markedly in NaIO3 group to 53% of normal group (P <0.01). No apparent difference was observed in naringenin+NaIO3 group. Four weeks later, the NaIO3 group fell to 37% of normal group (P <0.01), while the naringenin+ NaIO3 group fell to only 57% of normal group (P <0.01). There was a 52% reversal of the ERG C-wave by naringenin as compared to NaIO3 treated group (P <0.05). Two weeks and four weeks after laser treatment, naringenin reduced the CNV formation to 53% and 49% of control group (100%) measured by FA (P <0.01). Four weeks after laser treatment, naringenin reduced the CNV formation by 47% as compared to control group measured with flat mount(P <0.01). · CONCLUSION: Naringenin can significantly protect RPE from NaIO3 induced degeneration and also prevent CNV formation.     

Effect of Quercetin on Formation of Choroidal Neovascularization (CNV) in Age-related Macular Degeneration(AMD)

Purpose: Age-related macular degeneration (AMD) as a disease entity is "dry" at early stage and made up of two main components at late stage:atrophic AMD and exudative AMD. Quercetin acts as an anti-oxidant to protect retinal pigment epithelial cells (RPE) from damaged by oxidative stress, but its effect on formation of choroidal neovascularization (CNV)in AMD is unclear. The aim of this study is to investigate the effect of quercetin on the formation of CNV in AMD. Methods:The development of CNV induced by laser was detected by fluorescein angiography (FA). Colored microsphere technique was used to determine the choroidal blood flow in ocular hypertensive rabbit eyes.In in vitro studies, HUVECs were treated with NaIO3, H2O2 and NaN3 to induce oxidative cell damages. The effect of quercetin on various oxidationsinduced injuries in HUVECs was measured by MTT assay. HUVECs migration was assessed using a wound healing assay. Results:Quercetin significantly inhibited the formation of laser-induced CNV.The choroidal blood flow in rabbit eyes was significantly increased after quercetin instillation. In vitro results showed quercetin enhanced various oxidations-induced injuries in HUVECs and inhibited migration of HUVECs during wound healing. Conclusion: Quercetin inhibited the formation of CNV both in vivo and in vitro and increased choroidal blood flow.It could become a promising candidate for the treatment of AMD.