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.     

高氧诱导SD大鼠ROP动物模型的建立

目的:探讨用SD大鼠建立高氧诱导的早产儿视网膜病变(ROP)动物模型,为研究该病的发病机制及治疗提供理论基础。方法:新生SD大鼠24只分为2组,每组12只,实验组大鼠置于IPR-2小鼠隔离包,连续吸高浓度O27d后,再置于空气中饲养7d;对照组大鼠置于空气中饲养14d。通过视网膜铺片观察视网膜血管、组织切片HE染色观察视网膜新生血管、免疫组织化学观察VEGF和CD34蛋白的表达。结果:视网膜铺片显示实验组见大量无灌注区及新生血管芽,较对照组明显多。HE染色显示实验组内皮细胞数较对照组明显多。免疫组织化学表明CD34和VEGF在突破内界膜的细胞呈阳性表达。VEGF半定量测定显示实验组平均灰度值较对照组低,面密度值较对照组高。两组鼠生长发育均未见明显异常。结论:高氧诱导SD大鼠成功产生视网膜新生血管,可作为探究ROP疾病发生机制和治疗方法的可靠动物模型。     

精氨酸-谷氨酰胺在幼鼠早产儿视网膜病变模型中的应用(英文)

目的:观察精氨酸-谷氨酰胺(Arg-Gln)对早产儿视网膜病变动物模型视网膜新生血管的抑制作用。方法:48只7日龄的C57BL/6J新生鼠暴露在750mL/L高氧环境中5d,然后回到正常空气中建立早产儿视网膜病变的动物模型。在鼠龄12d时实验组(36只)新生鼠每天两次腹腔注射Arg-Gln(剂量分别为1.0,3.0,5.0g/kg,每组12只),连续注射5d;对照组(12只)每天两次腹腔注射PBS,连续5d。所有小鼠均于17d处死,视网膜铺片,ADP酶染色观察视网膜血管情况。HE染色,在光学显微镜下观察并计数突破视网膜内界膜的血管内皮细胞细胞核数目。Real-time RT-PCR方法测量每组视网膜VEGF mRNA水平。结果:与对照组相比,实验组以剂量依赖方式无灌注区面积和新生血管团逐渐减少;实验组中最大剂量组[5.0g/(kg·d)]突破内界膜的内皮细胞细胞核数目比对照组大约减少75%(P<0.01);实验组视网膜VEGF mRNA水平与对照组相比明显下降。结论:Arg-Gln能够有效抑制早产儿视网膜病变动物模型视网膜新生血管的生成,可能为临床提供一种预防和治疗早产儿视网膜病变安全有效的新方法。     

建立可定量的视网膜新生血管小鼠模型的探索

目的:建立可对视网膜新生血管进行定量研究的动物模型,为研究视网膜新生血管发生机制及治疗提供实验基础。方法:将32只7d龄C57BL/6J幼新生鼠在浓度为(75±2)%的高氧状态下生活5d,然后回到正常氧环境下,作为氧诱导模型组;另32只同日龄新生鼠置于正常空气环境中生活,作为正常对照组。采用荧光素血管灌注法视网膜铺片及作视网膜冰冻切片GSA(griffoniasimplicifolialectinB4)染色,了解视网膜血管的改变,定量计算新生鼠视网膜新生血管面积。结果:实验组新生鼠的视网膜血管在高氧中生长5d后,视网膜血管收缩、闭塞,出现大片无灌注区;回到正常空气环境下2d后,开始出现新生血管;回到空气中5d后,新生血管达到高峰。结论:该动物模型可重复性高,并可进行定量研究,是进行视网膜新生血管发生机制和药物治疗的合适模型。     

Cyclin D1在氧诱导视网膜病变大鼠视网膜中的表达

目的研究氧诱导视网膜病变(oxygen-induced retinopathy,OIR)中cyclin D1表达的变化对视网膜新生血管增殖的影响。方法 120只鼠龄7d SD大鼠随机分为实验组和对照组2组。实验组60只大鼠置于体积分数75%高氧环境5d建立OIR模型,对照组60只大鼠置于正常空气中。2组分别于生后第12天、第14天、第17天、第22天、第26天各取12只大鼠并均取双侧眼球,左眼做视网膜铺片ADP酶染色观察视网膜新生血管,免疫组化及免疫荧光观察cyclin D1的表达。右眼视网膜行Western blot检测cyclin D1蛋白表达情况。结果视网膜铺片ADP酶染色观察到大鼠生后第14天开始出现新生血管,至第17天新生血管反应最大,随后新生血管缓慢退化,在生后第26天血管结构趋于正常。免疫组化和免疫荧光显示大鼠生后第14天、第17天、第22天实验组视网膜cyclin D1呈高表达。Western blot结果显示cyclin D1蛋白的表达于生后第12天比对照组低,第14天开始增高,第17天达到最高,第26天降低,但高于对照组。Cyclin D1表达的变化符合新生血管增殖的过程。结论 cyclin D1的异常表达可能与视网膜新生血管增殖密切相关。     

小鼠视网膜新生血管模型的建立及特征

目的:建立可靠稳定的视网膜新生血管动物模型,为今后探究视网膜新生血管疾病的发生机制和治疗方法奠定模型基础。方法:将24只新生C57BL/6J小鼠随机分为正常组和模型组,每组各12只。将模型组小鼠于生后第7d置于750mL/L氧浓度环境,生后第12d返回正常空气中;正常组小鼠始终置于正常空气环境喂养。至小鼠生后第17d进行心脏荧光素灌注造影视网膜铺片以及眼球连续切片苏木精-伊红(H-E)染色,观测视网膜新生血管生成情况。结果:模型组小鼠心脏荧光素灌注造影结果显示视网膜中央区域呈无灌注缺血,另外视网膜血管有迂曲扩张、荧光渗漏等异常表现。眼球连续切片发现模型组小鼠突出视网膜内界膜的血管内皮细胞核数为48.65±6.24个/片/眼,而正常组小鼠平均为0.38±0.21个/片/眼,两组比较差异有显著统计学意义(P<0.01)。结论:氧诱导的视网膜缺血模型可成功诱导小鼠产生视网膜新生血管,可作为探究视网膜新生血管疾病发生机制和治疗方法的可靠动物模型。     

血管内皮生长因子在氧诱导小鼠新生血管中的表达及意义

目的探讨血管内皮生长因子（VEGF）在小鼠视网膜新生血管中的表达及意义。方法对新生c57BL／6N小鼠高氧后相对低氧饲养，诱导产生视网膜新生血管。在出生后12d、17d摘除眼球，应用RT-PCR，Western Blot以及视网膜血管荧光灌注造影技术检测全视网膜VEGF mRNA、蛋白表达水平以及视网膜新生血管的发生程度。结果视网膜血管造影显示高氧造成血管发育受限，相对低氧后产生新生血管。伴随新生血管的发生，VEGF mRNA升高2．3倍；VEGF蛋白含量也升高7．3倍。结论VEGF表达改变与新生血管发生成正相关，其升高也是造成病理性视网膜新生血管发生的机制之一。减少内源性VEGF表达可能成为治疗视网膜新生血管的新方法。     

代谢性酸中毒诱导新生大鼠视网膜 新生血管的实验研究

目的了解代谢性酸中毒诱导新生大鼠视网膜病变的发展进程，探讨其发生与血管内皮生长因子（VEGF）的关系。方法实验组新生Sprague-Dawley大鼠425只。从大鼠出生后第2天开始按535 mg/kg的剂量管饲NH₄Cl(浓度为50 mg/ml)，每天2次，管饲6 d，然后进入恢复期。另选150只新生大鼠未进行管饲，作为对照组。两组大鼠分别于出生后3、5、8、10、13、20 d处死。取出双眼进行视网膜铺片和二磷酸腺苷酶染色，对视网膜血管进行评估；用酶联免疫分析法进行VEGF的测定。结果实验组鼠在出生后3、5、8、10、13、20 d的新生血管（NV）发生率分别为0%、9%、26%、55%、19%、0%。出生后第3天，实验组鼠VEGF的蛋白水平［（101.1±14.2 ） pg/mg］比对照组［（133.2±15.9） pg/mg下降（P＝0.004），出后生第8天，实验组鼠VEGF蛋白水平［(98.4±19.2) pg/mg］比对照组［（78.1±8.7）pg/mg］升高（P=0.028)；出生后第5、10、13、20天，两组差异无统计学意义（P>0.05）。结论代谢性酸中毒可能损害了正在发育的视网膜血管而引起新生血管；酸中毒引起的视网膜新生血管与VEGF有关。(中华眼底病杂志,2005,21:296-299)     

组织蛋白酶B与血管内皮生长因子影响高氧诱导小鼠视网膜新生血管形成的实验研究

目的探讨组织蛋白酶B(cathepsin B)与血管内皮生长因子(VEGF)在小鼠视网膜新生血管形成中的影响。 方法选用清洁级C57BL/6J小鼠共44只,7~8日龄,雌雄不限。应用随机数字表法将其分为正常对照组和高氧处理组。其中,正常对照组11只小鼠(22只眼),高氧处理组33只小鼠(66只眼)。正常对照组小鼠在标准环境中生长;高氧处理组小鼠为建立视网膜新生血管动物模型,其环境除氧体积分数为(75±2)%,其他均相同,氧箱内饲养5 d后,将其返回到标准环境中。以视网膜表面出现新生血管簇、无灌注区,判定为造模成功。再应用随机数字表法将成模的小鼠随机分为模型对照组、实验对照组和实验组,每组11只小鼠(22只眼)。正常对照组和模型对照组不予任何药物干预,实验对照组和实验组分别给予玻璃体腔注射二甲基亚砜和溶于二甲基亚砜的CA-074 Me(cathepsin B抑制剂),继而在标准环境中饲养5 d。采用心腔注射荧光素标记葡聚糖,视网膜铺片法观察新生血管生成的情况;分别采用实时聚合酶链反应法和蛋白质印迹法检测小鼠视网膜组织中cathepsin B、VEGF信使核糖核酸(mRNA)相对表达量及蛋白的表达量。所有数据均以均数±标准差( ±s)表示。采用单因素方差分析比较各组总体差异,当差异有统计学意义时,再用SNK-q检验进行组间多重比较分析。 结果在荧光显微镜下,实验组较模型对照组和实验对照组视网膜血管走行相对清晰,未见明显的缺血无灌注区。各组cathepsin B与VEGF mRNA及蛋白的表达量总体存在差异,差异有统计学意义(F＝25.23,22.98,43.86,67.92;P<0.05)。模型对照组和实验对照组中cathepsin B与VEGF mRNA及蛋白的表达量均增高,而实验组中cathepsin B与VEGF的mRNA及蛋白的表达量则降低。实验组与正常对照组比较、实验对照组与模型对照组比较,差异均无统计学意义(P>0.05)。 结论CA-074 Me可抑制cathepsin B和VEGF的mRNA及蛋白的表达。cathepsin B和VEGF表达减少可抑制小鼠视网膜新生血管的形成,且cathepsin B与VEGF之间可能存在着双向调节的关系。     

PEDF抑制鼠视网膜新生血管的实验研究(英文)

目的:观察PEDF对氧诱导的血管增生性视网膜病变动物模型视网膜新生血管的抑制作用。方法:40只7日龄的C57BL/6J新生鼠暴露在750mL/L高氧环境中,然后回到正常空气中建立氧诱导的血管增生性视网膜病变的动物模型。随机取1眼为实验眼,在出氧箱时(12d)玻璃体腔注射PEDF,另1眼为对照组,玻璃体腔注射PBS。所有小鼠均于17d处死,视网膜铺片,ADP酶染色观察视网膜血管情况。HE染色,在光学显微镜下观察并计数突破视网膜内界膜的血管内皮细胞细胞核数目。结果:与对照组相比,实验组视网膜血管分布规则,未见明显的无灌注区形成;实验组突破内界膜的内皮细胞细胞核数目(10.18±1.74)比对照组(38.89±2.98)明显减少(P<0.01) ;组织切片未见视网膜毒性及炎症反应。结论:PEDF能够有效抑制视网膜新生血管的生成。     

Pathogenesis of Purtscher's retinopathy and Purtscher-like retinopathy

PURPOSE: The pathogenesis of Purtscher's retinopathy (PR) or Purtscher-like retinopathy (PIR) is illustrated on two case reports. MATERIAL AND METHODS: Five patients with PR or PIR were examined ophthalmologically. Fluorescein angiography, fundus photography, visual field testing, and electroretinography were also performed. RESULTS: In three cases, the PIR was observed after acute pancreatitis, in one case it arosed from cryoglobulinemy, because of hepatitis C, and in one case it was due to a classic PR after the thorax trauma. In the case of a slow resolution of retinal edema, atrophy of the retinal pigment epithelium and optic nerve, occurred. The therapy has been based on the internal medicine treatment of the causal disease and the administration of corticosteroids, to reduce retinal edema. CONCLUSIONS: PR and PIR are interdisciplinary diseases caused by microembolization of retinal vessels. If changes are intensive and long lasting, visual prognosis is poor.     

Radiation retinopathy: a mistaken diagnosis of hypertensive retinopathy

Radiation retinopathy is a vision‐threatening complication following therapeutic irradiation of ocular, orbital, facial, nasopharyngeal and cranial structures. It is characterised by a delayed onset, slowly progressive, occlusive retinal microangiopathy that develops several years after initial radiotherapy. We present the case of a 44‐year‐old man who developed radiation retinopathy, initially diagnosed as a case of hypertensive retinopathy, following irradiation of a nasopharyngeal carcinoma. A careful history along with classical clinical features and fundus fluorescein angiography helped establish the diagnosis.     

Solar retinopathy

319 patients with a solar retinopathy were seen in an eye clinic in Nepal within 20 months. All patients had either a positive history of sun-gazing or typical circumscribed scars in the foveal area. In more than 80% of the patients the visual acuity was 6/12 or better and did not deteriorate over time. 126 (40%) patients had a history of gazing at the sun during an eclipse, 33 (10%) were sun worshipers and 4 (1%) were in both categories. Three years later 29 patients were re-examined in a follow-up study. Only 16 had had visual disturbances directly after they had gazed into the sun. No colour vision defects were seen in any of the 44 affected eyes, when tested with Panel D 15, while four patients (6 eyes) had some uncertainty with the tritan plates of the Ishihara test charts. Metamorphopsia were recorded in 11 eyes. Five German patients with solar retinopathy were examined in more detail. Colour contrast sensitivity (CCS) was tested for the central and the peripheral visual field. CCS for tritan axis was raised in all patients for the central visual field, while it was normal for the peripheral visual field. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cartwheel retinopathy

A case of Valsalva retinopathy is presented, with a discussion of treatment options.     

Talc retinopathy

A 45 year old man presented with talc retinopathy that had been present for decades. The static nature of this condition indicates that, in the absence of ongoing intravenous drug use, ophthalmological follow up is not necessary     

Melanoma-associated retinopathy

A 63-year-old Taiwanese man with a history of cutaneous melanoma presented with a rapid onset of bilateral shimmering light and blurred vision. A fundoscopic examination was normal. However, visual field examination indicated generalized depression in both eyes. Scotopic rod-specific electroretinography (ERG) was undetectable and scotopic maximal combined-cone and rod-specific ERG showed the characteristics of negative ERG (a normal a-wave and a diminished b-wave, with the b-wave smaller than the a-wave), indicating dysfunction of the bipolar cells. Melanoma-associated retinopathy (MAR) was suspected and a systemic work-up gave a diagnosis of metastatic melanoma. This case shows the typical presentation of MAR. Greater awareness of MAR in patients with unexplained visual loss may help to identify an occult focus of metastatic melanoma.