年龄相关性黄斑变性患者黄斑区脉络膜新生血管特征分析

目的:探讨年龄相关性黄斑变性黄斑区脉络膜新生血管(CNV)的特征。方法:对41例(41只眼)年龄相关性黄斑变性患者CNV进行荧光素眼底血管造影(FFA)和相干光断层扫描(OCT)检查,并对两种不同的图像检查结果进行对比分析。结果:FFA发现典型CNV29眼,FFA早期黄斑区可见边界清晰的高荧光区,FFA晚期荧光扩大并增强;隐匿型CNV10眼,FFA早期可见边界欠清的强荧光,FFA晚期持续为荧光素渗漏或染色,边界多不清晰。OCT发现典型CNV33眼,显示CNV多位于视网膜神经上皮层下,呈边界清晰的纺锤形或圆形强反射,隐匿型CNV8眼,表现为视网膜色素上皮层平面不规则增强紊乱的反射信号。两种检查对黄斑区CNV的检出率对比差异无统计学意义(P0.05)。结论:结合FFA和OCT的图像特征对年龄相关性黄斑变性黄斑区脉络膜新生血管形态学改变进行对比分析,有助于明确病变的性质,为临床诊疗提供帮助。     

Amyloid beta在湿性年龄相关性黄斑变性患者中的表达及意义

目的:探讨Amyloid beta(Aβ)在湿性年龄相关性黄斑变性患者中的表达及意义。方法:2017年9月到2019年5月选择在本院诊治的湿性年龄相关性黄斑变性患者72例(72眼),检测血清Aβ含量,调查患者的一般资料、临床特征并进行相关性分析。结果:在72例患者中,平均血清Aβ含量为203.29±14.29 ng/mL,其中Aβ含量≥180 ng/mL 40例。Aβ阳性组的脉络膜厚度显著高于Aβ阴性组(P0.05)。Aβ阳性组的患眼动脉舒张末期血流速度和收缩期峰值血流速度都显著低于Aβ阴性组(P0.05)。Aβ阳性组的的性别、年龄、眼压、血压、体重指数、病程等与Aβ阴性组对比差异无统计学意义(P0.05)。直线相关性分析显示血清Aβ含量与脉络膜厚度、患眼动脉舒张末期血流速度和收缩期峰值血流速度都有显著相关性(P0.05),其中Aβ含量与脉络膜厚度呈正相关(r=0.566,P=0.003),Aβ含量与患眼动脉舒张末期血流速度和收缩期峰值血流速度呈负相关(r=-0.673,P=0.000;r=-0.455,P=0.010)。结论:Aβ在湿性年龄相关性黄斑变性患者中多呈现高表达状况,血清Aβ含量与脉络膜厚度、患眼动脉舒张末期血流速度和收缩期峰值血流速度相关,从而影响患者病情的进展。     

PDE2A通过影响脉络膜血管生成和NADPH氧化酶/ROS/NF-κB通路参与年龄相关性黄斑变性

摘要 目的：探究磷酸二酯酶2A（Phosphodiesterase 2A,PDE2A）在年龄相关性黄斑变性（Age-related macular degeneration,AMD）中的可能作用及其对脉络膜新生血管（Choroidal neovascularization,CNV）形成的影响。方法：通过qRT-PCR法检测36例湿性AMD患者（AMD组）和36例健康体检者（Healthy组）的血清PDE2A水平。将恒河猴脉络膜血管内皮细胞系（RF/6A）分为Normoxia组、Hypoxia组、Hypoxia+NC组、Hypoxia+PDE2A组。使用Lipofectamine2000分别将NC-shRNA和PDE2A-shRNA慢病毒转染入Hypoxia+NC组和Hypoxia+PDE2A组,转染后,根据分组情况,在RPMI 1640培养基中添加200 mM CoCl₂来对RF/6A细胞进行低氧处理。建立了激光诱导的CNV小鼠模型后,将36只建模成功的小鼠随机分为Model组、NC-shRNA组和PDE2A-shRNA组,每组12只。选取12只未建模的小鼠作为Control组。分别对NC-shRNA组和PDE2A-shRNA组小鼠玻璃体腔注射相应慢病毒,共治疗7 d。然后对小鼠进行眼底荧光血管造影（fundus fluorescein angiography,FFA）检查和眼球HE染色。使用相应试剂盒检测小鼠脉络膜组织中ROS、SOD、MDA的含量。通过qRT-PCR或Western blot检测RF/6A细胞或脉络膜组织中PDE2A、VEGFA、VEGFR2、HIF-1α、NOX2、NOX4和NF-κB p65的表达。结果：与Healthy组相比,AMD组患者的血清PDE2A水平显著升高（1.00±0.23 vs 3.09±1.46, P<0.001）。与Hypoxia组相比,Hypoxia+PDE2A组RF/6A细胞的闭合管腔数量减少（P<0.05）,PDE2A、VEGFA、VEGFR2和HIF-1?琢的mRNA和蛋白表达水平均降低（P<0.05）。与Model组相比,PDE2A-shRNA组CNV小鼠脉络膜病变明显减轻,血管生成和脉络膜增明显减少,CNV相对荧光强度降低（P<0.05）,脉络膜组织中的PDE2A、VEGFA、VEGFR2和HIF-1α的mRNA和蛋白表达水平均降低（P<0.05）。与Model组相比,PDE2A-shRNA组小鼠脉络膜组织中的ROS和MDA含量均降低,SOD含量升高（P<0.05）。与Model组相比,PDE2A-shRNA组小鼠脉络膜组织中的NOX2、NOX4和细胞核NF-κB p65蛋白相对表达量均降低（P<0.05）。结论：PDE2A通过影响脉络膜血管生成和NADPH氧化酶/ROS/NF-κB通路参与年龄相关性黄斑变性。     

新生血管性年龄相关性黄斑变性对抗VEGF治疗的耐药性危险因素分析

摘要 目的：探究新生血管性年龄相关性黄斑变性对抗血管内皮生长因子（VEGF）治疗的耐药性危险因素。方法：选取2019年5月至2021年5月于我院接受治疗的90例nAMD患者作为研究对象,进行VEGF治疗,根据患者的耐药情况将其分为耐药组（n=19）和非耐药组（n=71）,统计患者资料,对比分析产生耐药性的危险因素。结果：危险因素与AMD类型、PED、椭圆体带完整性、积液、出血、BCVA、眼压、CRT异常有关（P<0.05）,危险因素与性别和平均年龄无关（P>0.05）;Ⅰ型CNV患者在VEGF治疗前后CRT水平未出现显著变化（P>0.05）,II型CNV患者和混合型CNV患者治疗前后CRT水平均显著降低（P<0.05）。Logistic回归分析显示：最终进入主效应模型的因素即抗VEGF治疗的耐药性发生的多因素为积液、PED、出血、AMD类型。结论：新生血管性年龄相关性黄斑变性（nAMD）对抗VEGF治疗出现耐药性与积液、出血和AMD类型有关。     

姜黄素抑制小鼠脉络膜新生血管的实验研究

目的：观察姜黄素对激光诱导的小鼠脉络膜新生血管（choroidalneovascularization,CNV）形成的影响。方法：60只雄性C57BL／6小鼠,随机分为对照组、10mg／kg姜黄素治疗组、30mg／kg姜黄素治疗组,每组20只。采用激光诱导产生小鼠CNV模型。由光凝前3天开始,至光凝后14天,两个治疗组每天分别给予腹腔注射相应剂量的姜黄素,对照组腹腔注射二甲亚砜溶液（溶剂）。光凝后第3天通过免疫组化和ELISA检测血管内皮生长因子（vesselendothelialgrowthfactor,VEGF）的表达;第14天通过组织学检查以及荧光素标记的葡聚糖的血管灌注检测CNV的面积,荧光血管造影评价CNV的渗漏程度。结果：光凝后第14天,组织学检查显示姜黄素能够有效缩小激光诱导的CNV;荧光素标记的葡聚糖血管灌注后测量色素上皮-脉络膜铺片上CNV的面积,和对照相比,姜黄素能显著减小激光诱导的CNV的面积（P〈0．05）;荧光血管造影显示姜黄素能有效抑制CNV的渗漏（P〈O．05）。和10mg／kg姜黄素治疗组相比,30mg／kg姜黄素治疗组小鼠CNV面积缩小和渗漏程度减弱（P〈0．05）。光凝后第3天,VEGF免疫组化和ELISA结果显示姜黄素显著抑制色素上皮一脉络膜复合体中VEGF（P〈0．01）的表达,高刺量组有更强的抑制作用（P〈0．01）。结论：姜黄素可以有效地抑制小鼠CNV的形成,下调VEGF的表达可能是姜黄素抑制CNV的作用机制之一。因此我们推测姜黄素对并发CNV的AMD患者可能具有治疗作用。     

康柏西普与雷珠单抗治疗年龄相关性黄斑变性的临床研究

目的:探讨康柏西普联合雷珠单抗治疗年龄相关性黄斑变性(AMD)的临床疗效。方法:选择2015年6月到2016年10月我院收治的60例AMD患者,按随机数字表法分为对照组和治疗组。对照组患者给予康柏西普治疗,治疗组患者给予康柏西普联合雷珠单抗治疗,两组患者均治疗3个月。评价并比较两组患者临床疗效。统计并比较两组患者治疗后的视网膜渗漏总改善率。观察并比较两组患者治疗前后最佳矫正视力和黄斑视网膜厚度。结果:治疗后,治疗组患者的视力提高率为80.00%,明显高于对照组的55.00%,差异具有统计学意义(X~2=4.104,P=0.043)。治疗后,治疗组患者的视网膜渗漏总改善率为92.50%,明显高于对照组的70.00%,差异具有统计学意义(X~2=5.294,P=0.021)。治疗前,两组患者最佳矫正视力、黄斑视网膜厚度比较差异无统计学意义(P0.05);治疗后,两组患者最佳矫正视力均明显大于治疗前,黄斑视网膜厚度均明显小于治疗前,并且治疗组均明显优于对照组,差异均具有统计学意义(P0.05)。结论:康柏西普联合雷珠单抗治疗AMD的临床疗效显著,能够明显提高视力,减轻视网膜渗漏,值得在临床上推广应用。     

Wnt信号在年龄相关性黄斑变性组织和小鼠模型中的表达及作用

为了考察Wnt信号在年龄相关性黄斑变性组织和小鼠模型中的表达及作用,本研究采用免疫组织化学染色检测了10例干性年龄相关性黄斑变性病例和10例年龄相匹配的正常眼睛石蜡切片中LRP6和β-catenin的表达.通过氢醌诱导小鼠千性年龄相关性黄斑变性模型,然后应用Wnt信号通路抑制剂ICG001治疗模型小鼠4周.采用苏木精和...     

康柏西普与雷珠单抗对年龄相关性黄斑变性患者血清CRP、VEGF、眼压及视力的影响

目的:探讨康柏西普与雷珠单抗对年龄相关性黄斑变性(AMD)患者血清中C反应蛋白(CRP)、血管内皮生长因子(VEGF)水平、眼压(IOP)及视力的影响。方法:选取2015年4月到2016年5月在我院接受治疗的AMD患者70例作为研究对象,采用随机数字表法将所有患者分为观察组和对照组各35例,所有患者给予内眼手术标准玻璃体注射治疗,观察组给予康柏西普注射液1.5mg腔内注射,对照组给予雷珠单抗注射液0.5 mL腔内注射,对比两组患者治疗前、治疗后3d血清CRP、VEGF水平;记录并对比两组患者视力、IOP水平及并发症情况。结果:两组患者治疗后3d血清CRP、VEGF水平较治疗前下降,且观察组患者CRP、VEGF水平低于对照组(P0.05);两组患者治疗后3d视力最小视角对数(logMAR)及IOP较治疗前降低,且观察组患者视力logMAR及IOP水平明显低于对照组,差异均有统计学意义(P0.05)。治疗后3d,两组各出现1例玻璃体出血,并发症发生率比较差异无统计学意义(P0.05)。结论:相对于雷珠单抗,康柏西普更能有效降低AMD患者血清CRP、VEGF水平和IOP,提高患者视力,改善病情,值得临床推广应用。     

雷珠单抗与阿柏西普玻璃体腔注射治疗渗出性年龄相关性黄斑变性的疗效观察

目的:探究雷珠单抗与阿柏西普玻璃体腔注射治疗渗出性年龄相关性黄斑变性的临床疗效与安全性。方法:选取2015年5月～2018年5月我院收治的渗出性年龄相关性黄斑变性患者98例(98眼),采用随机数字表法将患者分为两组,A组玻璃体腔内缓慢注射0.05 m L(0.5 mg)雷珠单抗注射液,B组玻璃体腔内缓慢注射0.05 m L(2 mg)阿柏西普注射液。比较两组患者的视力改善情况、眼动脉血流动力学、黄斑中心凹视网膜厚度及不良反应的发生情况。结果:治疗前两组患者的视力比较无统计学差异(P0.05),治疗后,B组视力显著高于A组(P0.05);。两组患者治疗前后眼动脉血流动力学相关指标比较均无统计学差异(P0.05)。;两组患者治疗后的黄斑中心凹视网膜厚度均显著降低,且治疗3个月B组治疗3个月显著低于A组(P0.05);B组患者注射药物后结膜下大出血发生率显著低于A组(P0.05)。结论:阿柏西普可显著改善渗出性年龄相关性黄斑变性患者的视力,且安全性高,可能与其可显著改善患者的黄斑中心凹视网膜厚度有关,且不良反应发生率低,安全性好。     

PCL/β-TCP调控巨噬细胞极化对成血管效应的影响

目的:探究生物可降解材料聚己内酯/β-磷酸三钙(PCL/β-TCP)通过调控巨噬细胞极化对骨组织内血管生成的作用,为其临床应用提供依据.方法:采用3D打印技术制备试样并加以表征.体内实验采用模型为SD大鼠股骨远端植入模型.双侧植入PCL/β-TCP支架后采取免疫荧光染色观察支架内部成血管标记物CD31的表达差异,并采用...     

A new Chlorin formulation promotes efficient photodynamic action in choriocapillaris of rabbit’s eyes

Age-related macular degeneration (AMD) as well as other choroidal diseases, demand novel therapeutic methods. Photodynamic therapy (PDT), which uses light and photosensitizer (PS) to cause specific vascular occlusion in the macula, is an interesting alternative. The only drug approved for the PDT treatment of AMD (Verteporfin) has a natural tendency to aggregate, demanding an expensive separation procedure during purification. We report a novel and affordable PS that is intrinsically protected against aggregation, the Monomeric Chlorin at High Concentration (MCHC-Chlorin), whose liposomal formulation was developed to provoke effective photodynamic action on the choroidal vasculature. Our report starts by stablishing the conditions to allow the efficient synthesis of MCHC-Chlorin in high yields (92%). We then tested the light stimulated occlusion of choriocapillary vessels in rabbit’s eyes induced by the two MCHC-Chlorin isomers, which are directly obtained from the synthetic route. The PS formulation was infused in the rabbit’s ear vein and eyes were immediately irradiated at 650?nm. Indirect ophthalmoscopy, fundus photography, fluorescein angiography and histopathological evaluations were used to evaluate levels of photo-thrombosis and collateral damage. Choriocapillary occlusion was achieved in all treated rabbits’ eyes, while retina and sclera were completely preserved. There was no photochemical reaction in none of the eyes that received LASER without PS. Both MCHC-Chlorin isomers were separately tested and exhibited similar positive results with no systemic toxicity. Therefore, PDT occurred equally well in all treated eyes and none of the controls showed any effect in the ophthalmological exams. MCHC-Chlorin offers great potential and should be further studied as an alternative drug for choroidal diseases.     

Short interfering RNA (siRNA), a novel therapeutic tool acting on angiogenesis

The formation of new blood vessels, uncontrolled cell expansions and invasions are the common feature of cancer, neovascular inflammatory and ocular diseases, such as age-related macular degeneration (AMD). Short interfering RNA (siRNA) and short-hairpin RNA (shRNA) have recently helped extend our understanding of the mechanisms regulating angiogenesis and tumor developments. Moreover, the early success of these tools has reinforced the therapeutic hopes of preventing endogenous or exogenous gene translation. In vivo experiments using several animal tumor models and human pre-clinical trials augured many benefits to control protein expression and cell signaling. The high specificity of siRNA and shRNA to target a protein is crucial to design a new generation of therapeutic agents. At the present, several investigations are focused on the understanding of both gene function and the proof-of-concept for siRNA-mediated anti-angiogenesis. Taken together, in vitro and in vivo studies shed light on the efficiency of siRNA as a new alternative therapeutic agent.     

Pegaptanib in the treatment of wet, age-related macular degeneration

Age-related macular degeneration (AMD) is a major cause of severe visual loss worldwide. Neovascular (wet) AMD accounts for 90% of the visual loss associated with the disorder and vascular endothelial growth factor (VEGF) has been shown to play a major role in neovascularization and vascular permeability, the major causes of visual loss in AMD, making it an ideal target for therapeutic intervention. To utilize this strategy, pegaptanib, an aptamer that specifically binds to and blocks VEGF165, the VEGF isoform primarily responsible for abnormal vascular growth and permeability in AMD, was developed. Following encouraging preclinical trials, clinical trials showed that pegaptanib stabilized vision and reduced the risk of severe visual loss in the majority of patients with AMD, with some patients showing visual improvement. Pegaptanib has maintained a good safety profile with only occasional adverse effects. Even greater success was achieved when pegaptanib was used in combination with another therapeutic strategy, such as photodynamic therapy or bevacizumab, a pan isoform VEGF inhibitor. Further investigation of pegaptanib for the therapy of wet AMD, particularly in combination with other modes of therapy, should be encouraged.     

Genetic analysis of typical wet-type age-related macular degeneration and polypoidal choroidal vasculopathy in Japanese population

Age-related macular degeneration (AMD) is a common cause of blindness in the elderly. Caucasian patients are predominantly affected by the dry form of AMD, whereas Japanese patients have predominantly the wet form of AMD and/or polypoidal choroidal vasculopathy (PCV). Although genetic association in the 10q26 (ARMS2/HTRA1) region has been established in many ethnic groups for dry-type AMD, typical wet-type AMD, and PCV, the contribution of the 1q32 (CFH) region seem to differ among these groups. Here we show a single nucleotide polymorphism (SNP) in the ARMS2/HTRA1 locus is associated in the whole genome for Japanese typical wet-type AMD (rs10490924: , OR = 4.16) and PCV (rs10490924: , OR = 2.72) followed by CFH (rs800292: , OR = 2.08; , OR = 2.00), which differs from previous studies in Caucasian populations. Moreover, a SNP (rs2241394) in complement component C3 gene showed significant association with PCV (, OR = 3.47). We conclude that dry-type AMD, typical wet-type AMD, and PCV have both common and distinct genetic risks that become apparent when comparing Japanese versus Caucasian populations.

Electronic supplementary material

The online version of this article (doi:10.1007/s12177-009-9047-1) contains supplementary material, which is available to authorized users.     

Synthesis and structural characterization of carboxyethylpyrrole-modified proteins: mediators of age-related macular degeneration

Protein modifications in which the ε-amino group of lysyl residues is incorporated into a 2-(ω-carboxyethyl)pyrrole (CEP) are mediators of age-related macular degeneration (AMD). They promote both angiogenesis into the retina (‘wet AMD’) and geographic retinal atrophy (‘dry AMD’). Blood levels of CEPs are biomarkers for clinical prognosis of the disease. To enable mechanistic studies of their role in promoting AMD, for example, through the activation of B- and T-cells, interaction with receptors, or binding with complement proteins, we developed an efficient synthesis of CEP derivatives, that is especially effective for proteins. The structures of tryptic peptides derived from CEP-modified proteins were also determined. A key finding is that 4,7-dioxoheptanoic acid 9-fluorenylmethyl ester reacts with primary amines to provide 9-fluorenylmethyl esters of CEP-modified proteins that can be deprotected in situ with 1,8-diazabicyclo[5.4.0]undec-7-ene without causing protein denaturation. The introduction of multiple CEP-modifications with a wide variety of CEP:protein ratios is readily achieved using this strategy.