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。     

黄酮对视网膜色素上皮细胞氧化损伤的保护作用

目的:观察黄酮对氧化剂所诱导的视网膜色素上皮细胞(RPE)的保护作用。方法:在体研究中,预先给予5g/L黄酮滴眼液(3次/d),1wk后舌下静脉注射NaIO3诱导大鼠RPE变性,在2和4wk末,采用视网膜电图(ERG)测量C波。离体研究中,采用缺氧、H2O2、NaN3和t-BHP诱导RPE细胞损伤,并用MTT法检测细胞的存活率。结果:ERG的C波结果表明,第4wk末,黄酮抑制了由NaIO3诱导的大鼠RPE变性。离体研究结果表明,黄酮对多种氧化剂所诱导的RPE细胞损伤具有保护作用。结论:黄酮对氧化诱导的在体和离体视网膜色素上皮细胞均具有保护作用。     

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。     

色素家兔脉络膜新生血管中结缔组织生长因子表达

目的:通过激光诱导色素家兔脉络膜新生血管(choroidalneovascularization,CNV)模型,探讨结缔组织生长因子(connectivetissuegrowthfactor,CTGF)在CNV形成中的作用。方法:用高强度半导体泵浦的倍频Nd:YAG532nm绿激光光凝兔视网膜后极部,诱发CNV。术后1,3,4wk用原位杂交方法检测CTGFmRNA在CNV中的表达。结果:光凝后1wk,CNV膜开始形成,CTGFmRNA主要表达于CNV及其附近;光凝后3,4wk,CTGFmRNA仍明显表达,并有增强的趋势。结论:结缔组织生长因子可能参与了实验性CNV的发生发展。     

血管内皮生长因子在实验性脉络膜新生血管生成中表达的时相变化

目的:探讨血管内皮生长因子(VEGF)在激光诱导的大鼠脉络膜新生血管(CNV)生成中表达的时相变化。方法:应用532nm倍频激光对30只有色大鼠的实验眼进行视网膜光凝,建立CNV模型,分别于光凝后3d;1,2,3和4wk行荧光素眼底血管造影(FFA)后处死大鼠,摘除眼球,制作组织病理学标本,原位杂交法检测VEGFmRNA表达,免疫组织化学法和免疫荧光染色法检测VEGF蛋白的表达。结果:正常大鼠视网膜神经节细胞层、内核层、色素上皮层、视网膜及脉络膜血管内皮细胞中均有VEGFmRNA和蛋白表达。光凝后3d,Ⅷ因子阳性的内皮细胞由破裂的Bruch膜长入视网膜下,但尚未形成CNV;光凝后7d,Ⅷ因子阳性的内皮细胞围成的血管样结构,FFA检查可见相应区域有圆盘状荧光素渗漏,表明CNV生成。同时发现,光凝后3d,在光凝区外核层缺损处,增生和移行的色素上皮细胞、色素性巨噬细胞、成纤维细胞和增生的内皮细胞中VEGFmRNA和蛋白的表达呈强阳性;CNV中VEGFmRNA和蛋白在1wk时表达最强(P<0.01),2wk后其表达迅速下降。结论:VEGF的表达早于CNV的生成,提示其潜在的重要作用。     

TGF-β在氩激光诱导大鼠脉络膜新生血管中的表达

目的:评价血管内皮转化生长因子-β(transforming growth factor-beta,TGF-β)在氩激光诱导RWH大鼠脉络膜新生血管(CNV)形成过程中的表达及其意义。方法:RWH大鼠21只,随机分成3组,每组7只。单眼视网膜氩激光光凝诱导CNV形成后,分别在光凝后1,2,3wk摘除双眼球,应用免疫组织化学技术对光凝区进行因子Ⅷ相关抗原(FⅧR∶Ag)的检测以观察CNV形成,并检测CNV形成过程中TGF-β的表达,分析其变化趋势。结果:FⅧR∶Ag免疫组织化学检查结果显示,光凝后1wk开始形成CNV,3wk达到高峰。正常RWH大鼠视网膜中,TGF-β在视网膜神经节细胞层、内核层、色素上皮层、视网膜和脉络膜血管内皮细胞表达。视网膜光凝后,除上述部位外,TGF-β在外核层和脉络膜的光凝损伤区阳性表达。光凝后1～3wk,光斑区内FⅧR∶Ag和TGF-β阳性染色密度逐渐增加(P<0.05),FⅧR∶Ag与TGF-β阳性染色密度呈正相关(r=0.83,P<0.05)。结论:CNV形成与TGF-β表达正相关,TGF-β表达增多可能是CNV形成和增殖的主要原因之一。     

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.     

外源性VEGF及Ang-1对大鼠CNV的作用及机制探讨

目的:研究细胞因子VEGF,Ang-1对大鼠脉络膜新生血管的形成、分化以及成熟过程的影响及机制。方法:采用激光光凝诱导建立大鼠CNV动物模型后,分别在不同时期玻璃体腔注射VEGF,Ang-1,通过其眼底荧光造影与病理切片的变化观察上述因子的作用。结果:血管生成素-1组比对照组晚期渗漏明显减少,VEGF组较对照组晚期渗漏明显增加;组织病理切片显示实验组瘢痕形成,VEGF组和对照组可见肉芽组织。结论:血管生成素-1能有效抑制脉络膜新生血管的生成,促进新生的脉络膜新生血管成熟,促进瘢痕形成。     

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。     

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.     

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.     

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.     

Effects of flavone on the oxidation-induced injury of retinal pigment epithelium cells

AIM: To investigate the effect of flavone on oxidation- induced injury in retinal pigment epithelium cells. METHODS: In in vivo studies, NaIO3-induced RPE degene- ration in rat eyes was treated with 0.5% flavone eye drops 3 times a day for 1 week before and 4 weeks after NaIO3 injection. At the end of 2 and 4 weeks, all rats were measured c-wave by electroretinogram (ERG). In in vitro studies, ARPE-19 cells were treated with hypoxia, H2O2, NaN3 and t-BHP to induce cell damages. MTT assay was used to measure the viable cells. RESULTS: The ERG c-wave results showed that flavone reversed NaIO3-induced injury at the end of 4 weeks. In vitro results showed flavone reversed the various oxidants-induced injuries in RPE cells. CONCLUSION: Flavone could prevent the RPE from oxidation- induced injury both in vivo and in vitro.     

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).     

Targeted delivery of anti-angiogenic agent TNP-470 using water-soluble polymer in the treatment of choroidal neovascularization.

PURPOSE: The conjugation of drugs with water-soluble polymers such as poly(vinyl alcohol) (PVA) tends to prolong the half-life of drugs and facilitate the accumulation of drugs in tissues involving neovascularization. The purpose of this study was to evaluate the effect of TNP-470-PVA conjugate on the proliferation of endothelial cells in vitro and on experimental choroidal neovascularization (CNV) in vivo. METHODS: TNP-470 was conjugated in PVA by a dimethylaminopyridine-catalyzed reaction. The effects of TNP-470-PVA and free TNP-470 on the proliferation of human umbilical vein endothelial cells (HUVECs) and bovine retinal pigment epithelial cells (BRPECs) were evaluated by the tetrazolium-based colorimetric assay (XTT assay). Experimental CNV was induced by subretinal injection of gelatin microspheres containing basic fibroblast growth factor, into rabbits. Thirty rabbits were intravenously treated either with TNP-470-PVA (n = 8), free TNP470 (n = 5), free PVA (n = 5), or saline (n = 12) daily for 3 days, 2 weeks after implantation of gelatin microspheres. Fluorescein angiography was performed to detect the area with CNV, and the evaluation was made by computerized measurement of digital images. These eyes were also examined histologically. To observe the accumulation of conjugate, 3 rabbits with CNV received rhodamine B isothiocyanate-binding PVA (RITC-PVA), and the lesion was studied 24 hours later by fluorescein microscopy. RESULTS: The TNP-470-PVA inhibited the growth of HUVECs, similar to that of free TNP-470. The BRPECs were less sensitive to TNP-470-PVA than were the HUVECs. TNP-470-PVA significantly inhibited the progression of CNV in rabbits (P = 0.001). Histologic studies at 4 weeks after treatment demonstrated that the degree of vascular formation and the number of vascular endothelial cells in the subretinal membrane of the eyes treated with TNP-470-PVA were less than those of the control eyes. RITC-PVA remained in the area with CNV 24 hours after administration. CONCLUSIONS: These results suggest that TNP-470-PVA inhibited the proliferation of HUVECs more sensitively than that of BRPECs, and the targeted delivery of TNP-470-PVA may have potential as a treatment modality for CNV.     

Effect of focal X-ray irradiation on experimental choroidal neovascularization.

PURPOSE: Radiation therapy has been used to treat choroidal neovascularization (CNV) in patients with age-related macular degeneration. The in vivo effect of applying focal x-ray irradiation to the eye of rabbits with experimental CNV was investigated. METHODS: CNV was induced in the rabbit eyes by subretinal implantation of gelatin hydrogel microspheres impregnated with basic fibroblast growth factor. Three weeks after implantation, 17 of 34 eyes with CNV lesions accompanied by fluorescein leakage were irradiated with a single dose of 20 Gy; the other 17 eyes were not irradiated and served as the controls. The eyes were examined before irradiation and 1, 2, and 4 weeks after irradiation, by indirect ophthalmoscopy and fluorescein angiography. The degree of a decreasing amount of fluorescein leakage from the CNV lesions after irradiation was graded using a computerized image analysis system and was compared in the irradiated and nonirradiated eyes. These eyes were also examined histologically and immunohistochemically. RESULTS: Fluorescein leakage from the CNV lesions had significantly decreased in the eyes irradiated with 20 Gy compared with the control eyes, throughout the study period (P < 0.05). Histologic and immunohistochemical studies at 4 weeks after irradiation demonstrated that the degree of vascular formation and the number of vascular endothelial cells in the subretinal membrane of the irradiated eyes were less than those of the control eyes. CONCLUSIONS: Focal x-ray irradiation at the ocular region effectively reduced experimental CNV activity. These results support the possibility that radiation therapy may be beneficial in treating CNV.