单核细胞趋化蛋白-1在早期糖尿病大鼠视网膜中的表达及意义

目的　探讨单核细胞趋化蛋白 1 (MCP 1 )在早期糖尿病大鼠视网膜中的表达及其与糖尿病视网膜病变(DR)的关系。 方法　Wistar大鼠随机分为正常对照组 (NC组 )和糖尿病模型组 (DM组 ),每组 12只。大鼠腹腔注射链脲佐菌素(STZ)诱发糖尿病模型。8周时全部处死,采用Western免疫印迹法和免疫组织化学法,分析MCP 1在早期DM大鼠视网膜中的表达,对免疫印迹结果采用计算机图像分析系统处理。 结果　8周时,免疫印迹结果显示视网膜中MCP 1的表达DM组明显高于NC组(P<0 01),免疫组织化学法显示 8周时DM大鼠视网膜表面血管、内核层均可见MCP 1阳性着染细胞。 结论　MCP 1在视网膜组织中的表达与DR的发生有密切关系。     

单核细胞趋化蛋白-1在实验性糖尿病大鼠视网膜中的表达及意义

目的 研究单核细胞趋化蛋白（monocyte chemoattractant protem-1,MCP-1）在不同病程实验性糖尿病大鼠视网膜的表达及探讨MCP-1与糖尿病视网膜病变发生发展的关系.方法 Wistar大鼠64只随机分为正常对照组（NC组）16只和糖尿病造模组（DM组）48只,DM组腹腔注射链尿佐菌素诱发糖尿病,将造模成功DM组糖尿病大鼠随机分为2周、4周、12周和20周组,每组各12只.到各时间点后处死,采用Western Blotting和免疫组织化学法,分析MCP-1在不同病程糖尿病造模大鼠视网膜中的表达,对免疫印迹结果采用计算机图像分析系统处理.结果 免疫印迹结果显示：正常大鼠视网膜中MCP-1的表达非常微弱,糖尿病诱导成功后2周MCP-1蛋白表达比正常组明显增加（P＜0.05）,并随时间的延长表达逐渐增加,到12周时达最高.免疫组织化学结果显示：正常大鼠视网膜各层均无MCP-1表达,大鼠糖尿病诱导成功后2周,视网膜表面血管即有MCP-1阳性细胞表达,以后随病程延长而表达增多.结论 MCP-1在视网膜的表达与糖尿病视网膜病变的发生密切相关.     

白细胞介素-1β诱导大鼠视网膜单核细胞趋化蛋白-1mRNA表达的研究

目的探讨白细胞介素-1β(IL-1β)对大鼠视网膜单核细胞趋化蛋白(MCP)-1mRNA表达的诱导作用. 方法 24只SD大鼠,右眼为实验眼,左眼为对照眼.实验眼玻璃体内注入IL-1β 250U/5μl,对照眼注入等量生理盐水(PSS).注射后4,24h取视网膜组织,提取总RNA应用反转录-聚合酶链反应(RT-PCR)检测视网膜MCP-1mRNA表达. 结果眼内IL-1β注射后4h实验眼视网膜MCP-1mRNA表达水平较对照眼明显增加;24h实验眼视网膜MCP-1mRNA水平与对照眼相近. 结论视网膜MCP-1mRNA可被眼内IL-1β诱导表达,MCP-1可能在IL-1β诱发的眼内炎症反应中发挥作用.     

单核细胞趋化蛋白-1在氧诱导视网膜病变新生小鼠模型中的表达

背景 单核细胞趋化蛋白-1( MCP-1)是趋化因子家族中的主要成员之一,在肿瘤、炎症和糖尿病视网膜病变(DR)等新生血管性疾病中起着重要的作用,但关于MCP-1在氧诱导视网膜病变(OIR)发生过程中的表达及其作用的研究鲜有报道. 目的 观察OIR小鼠视网膜中MCP-1的表达,探讨MCP-1在视网膜血管发育和视网膜新生血管形成过程中的作用. 方法 SPF级C57BL/6J小鼠120只,按随机数字表法随机分为OIR组和正常对照组,每组60只.OIR组将出生后7d的新生小鼠在体积分数75％氧环境下饲养5d,然后返回正常大气环境中;正常对照组小鼠始终置于正常大气环境中饲养.OIR组和正常对照组分别在出生后5、7、12、14、17、21 d随机抽取10只小鼠,摘除右眼球,采用免疫组织化学法检测MCP-1蛋白在小鼠视网膜中的表达情况,采用逆转录-聚合酶链反应(RT-PCR)法检测MCP-1 mRNA在小鼠视网膜中的表达. 结果 正常对照组小鼠在出生后的第5天即有MCP-1在视网膜的内核层和节细胞层表达,12d时表达MCP-1的阳性细胞数达到高峰,其他日龄时呈基线表达.OIR组12d时表达MCP-1的阳性细胞数轻度增多,14d时阳性细胞数显著增多,之后迅速下降至正常水平.正常对照组在5d时可检测到MCP-1 mRNA表达,12d时显著上调,之后随小鼠日龄的增加,MCP-1 mRNA表达迅速下降,14、17、21 d表达基本呈基线水平.OIR组12 d时,MCP-1 mRNA较正常对照组下降,在新生血管形成关键期,即14 d时表达显著上调,之后迅速下降至正常水平.OIR组和正常对照组间比较采用完全随机分组的两因素方差分析,F分组=5.230,P=0.028;F日龄=6.898,P=0.001.结论 小鼠视网膜发育过程始终伴随着MCP-1的表达,MCP-1的表达上调可能与小鼠视网膜血管发育和OIR模型中视网膜新生血管的形成密切相关.     

白细胞介素-1β诱导大鼠视网膜单核细胞趋化蛋白-1mRNA表达的研究

目的 探讨白细胞介素－１β（ＩＬ－１β）对大鼠视网膜单核细胞趋化蛋白（ＭＣＰ）－１ｍＲＮＡ表达的诱导作用。方法 ２４只ＳＤ大鼠,右眼为实验眼,左眼为对照眼。实验眼玻璃体内注入ＩＬ－１β ２５０ Ｕ／５μｌ,对照眼注入等量生理盐水（ＰＳＳ）。注射后４,２４ｈ取视网膜组织,提取总ＲＮＡ应用反转录－聚合酶链反应（ＲＴ－ＰＣＲ）检测视网膜ＭＣＰ－１ｍＲＮＡ表达。结果 眼内ＩＬ－１β注射后４ｈ实验眼视网膜ＭＣＰ－１ｍＲＮＡ表达水平较对照眼明显增加;２４ｈ实验眼视网膜ＭＣＰ－１ｍＲＮＡ水平与对照眼相近。结论 视网膜ＭＣＰ－１ｍＲＮＡ可被眼内ＩＬ－１β诱导表达,ＭＣＰ－１可能在ＩＬ－１β诱发的眼内炎症反应中发挥作用。     

房水和血清中单核细胞趋化蛋白-1和巨噬细胞游走抑制因子变化与2型糖尿病视网膜病变的关系

背景研究发现,巨噬细胞和白细胞等炎性细胞参与糖尿病视网膜病变（DR）的发生发展,有多种细胞因子在DR的发生发展过程中发挥促进作用,但DR患者的房水及血清中单核细胞趋化蛋白-1（MCP-1）和巨噬细胞游走抑制因子（MIF）水平的变化与DR病程的关系尚不完全清楚。目的探讨2型糖尿病患者房水中MCP-1和MIF的水平与DR病程的关系。方法纳入2010年9月至2011年6月在北京世纪坛医院眼科和内分泌科确诊的2型糖尿病合并白内障并已行白内障超声乳化手术或超声乳化联合玻璃体切割术患者80例,根据眼底情况分为无DR（NDR）组20例、非增生型DR（NPDR）组38例和增生型DR（PDR）组22例,并收集同期的非糖尿病白内障手术患者即对照组26例,各组患者年龄、性别分布的差异均无统计学意义。所有患者于术中抽取未稀释的房水0．1ml,分别采用ELISA法检测房水中MCP-1和MIF的质量浓度并进行组间比较。结果PDR组、NPDR组、NDR组、对照组房水中平均MCP-1的质量浓度分别为（1660．78±562．98）、（1463．26±623．41）、（686．76±186．16）、（494．35±148．59）ng／L,总体比较差异有统计学意义（F=37．968,P=0．000）,对照组与NDR组以及NPDR组与PDR组房水中MCP-1的质量浓度比较差异均无统计学意义（P=0．169、0．117）;NDR组以及NPDR组与PDR组房水中MCP-1的质量浓度均明显高于对照组,差异均有统计学意义（P=0．000）。PDR组、NPDR组、NDR组、对照组患者房水中平均MIF的质量浓度分别为（6．85±1．99）、（3．56±0．90）、（1．10±0．48）、（0．86±0．46）μg／L,总体比较差异有统计学意义（F=144．502,P=0．000）;对照组与NDR组房水中MIF的质量浓度比较差异无统计学意义（P=0．475）;其余各组间两两比较差异均有统计学意义（P=0．000）。所有受检者房水中MCP-1与MIF质量浓度变化呈正相关（r=0．564,P=0．000）。PDR组、NPDR组、NDR组血清中MCP-1和MIF质量浓度较对照组均有所增加,但4个组间MCP-1和MIF质量浓度的总体比较差异均无统计学意义（F=2．158、0．813,P〉0．05）。结论糖尿病患者房水中MCP一1和MIF与DR的严重程度密切相关,MCP一1和MIF在DR的损伤机制中有协同作用。     

白细胞介素—1β诱导大鼠视网膜单核细胞趋化蛋白—1mRNA表达的研究

目的 探讨白细胞介素-1β（IL-1β）对大鼠视网膜单核细胞趋化蛋白（MCP）-1mRNA表达的诱导作用。方法 24只SD大鼠,右眼为实验眼,左眼为对照眼,实验眼玻璃体内注入IL-1β250U/5μl,对照眼注入等量生理盐水（PSS）。注射后4,24h取视网膜组织,提取总RNA应用反转录-聚合酶链反应（RT-PCR）检测视网膜MCP-1mRNA表达。结果 眼内IL-1β注射后4h实验眼视网膜MCP-1mRNA表达水平较对照h实验眼视网膜MCP-1mRNA水平与对照眼相近。结论 视网膜MCP-1mRNA可视眼内IL-1β诱导表达,MCP-1可能在IL-1β诱发的眼内炎症反应中发挥作用。     

单核细胞趋化蛋白-1和基质金属蛋白酶-2在增生性玻璃体视网膜病变增生膜中的表达

目的观察单核细胞趋化蛋白-1(monocyte chemotac ticprotein-1，MCP-1)和基质金属蛋白酶-2(matrix metalloproteinase-2，MMP-2)在增生性玻璃体视网膜病变(PVR)增生膜中的表达。方法 玻璃体手术取出的PVR增生膜24例，其中C级膜11例，D级膜13例，用免疫组织化学方法检测MCP-1和MMP-2的表达。结果在全部24例增生膜中，MCP-1和MMP-2均有表达，MCP-1阳性表达8例，强阳性表达16例；MMP-2阳性表达7例，强阳性表达17例。结论MCP-1和MMP-2均存在于PVR增生膜中，提示2者在PVR增生膜的形成和PVR发生发展过程中起重要作用。     

单核细胞趋化蛋白-1在增生型糖尿病视网膜病变患者玻璃体中的表达

研究表明,炎症反应与糖尿病视网膜病变(DR)的发病机制密切相关[1].趋化因子是炎症反应的重要因子,可以诱导炎性细胞发生迁移聚集,在炎症性疾病的发生发展中起重要作用.单核细胞趋化蛋白-1(MCP-1)是单核细胞及巨噬细胞的强激活剂,能使大约30％的单核细胞发生迁移[2,3].为了进一步探讨MCP-1与DR发生发展的关系,我们观察了一组增生型DR(PDR)患者玻璃体中MCP-1的表达.现将结果报道如下.     

单核细胞趋化蛋白-1在大鼠脉络膜新生血管中的表达

目的观察单核细胞趋化蛋白-1（MCP-1）在氪激光诱导的BN大鼠脉络膜新生血管（CNV）中的表达。方法用氪激光对48只健康雄性BN大鼠的一侧眼进行视网膜光凝以建立CNV动物模型,光凝后连续8周每周对模型鼠行荧光素眼底血管造影（FFA）检查,每周检查后各摘除6只模型眼制备眼球壁切片。依据苏木精-伊红染色测量CNV面积,应用免疫组织化学法测定MCP-1、Ⅷ因子相关抗原（FⅧ-Rag）蛋白在CNV中的表达。结果视网膜光凝后1～8周,CNV面积、荧光素渗漏、FⅧ-RAg和MCP-1在CNV中的表达量随着时间的延长逐渐增加,差异均有统计学意义（F=194.43,P〈0.01;F=178.84,P〈0.01;F=965.15,P〈0.01;F=1400.71,P〈0.01）。CNV面积与荧光素渗漏量（r=0.886,P〈0.05）、CNV面积与FⅧ-RAg表达量（r=0.975,P〈0.05）、CNV面积与MCP-1表达量（r=0.956,P〈0.05）、MCP-1表达量与FⅧ-RAg表达量（r=0.961,P〈0.05）、MCP-1表达量与荧光素渗漏量（r=0.914,P〈0.05）均呈正相关。结论 MCP-1的表达随CNV面积及荧光素渗漏量的增加而增强,可能在CNV形成和发展过程中起重要作用。     

蚕食式膜切除术在糖尿病视网膜病变中的应用

目的:从术中术后并发症的发生率评价蚕食式膜切除术在糖尿病视网膜病变中的作用。方法:对比分析了行普通撕膜技术的31例(34只眼)和行蚕食式膜切除术的52例(58只眼)术中术后并发症的发生率。结果:蚕食式膜切除术组发生医源性视网膜裂孔2只眼,普通手术组7例(P<0.05);蚕食式膜切除术组术中出血4只眼,普通手术组15只眼(P<0.01);蚕食式膜切除术组术后有6只眼发生Ⅲ级反应,普通手术组有5只眼(P>0.05)。结论:糖尿病视网膜病变中使用蚕食式膜切除术剥膜比较安全,能减少手术中及手术后并发症。     

Overview of Epidemiologic Studies of Diabetic Retinopathy

Diabetic retinopathy has been an important cause of blindness in young and middle age adults in the United States. Epidemiologic studies have quantitated the risk and have described potentially causal factors associated with many ocular complications of diabetes and other facets of this disease. A review of recent advances in diagnosis, treatment, temporal trends, and health care for diabetic retinopathy was conducted. Since the early 1980's, there have been studies of the variability of diabetic retinopathy in populations around the world and subpopulations in the United States which have demonstrated the high prevalences and incidences of this condition. Observational studies and clinical trials have documented the importance of glycemic and blood pressure control in the development and progression of this disease. There are some differences in the importance of confounders in different populations. Epidemiologic data have helped understand the importance of health care and health education in prevention and treatment of this condition. Observational studies have documented the importance of this disease on quality of life. Although there have been advances in understanding the distribution, causes, and severity of diabetic retinopathy, this is ever changing and requires continued monitoring. This is important because the increasing burden of diabetes will place a greater burden on the population and the medical care systems that will be caring for them.     

阿斯匹林防治糖尿病性视网膜病变最佳药物剂量探讨

应用放射免疫法和比浊法测定31例单纯型糖尿病性视网膜病变患者分别服用25mg／日、50mg、日、100mg／日剂量的阿斯匹林各一个疗程对血浆血栓素水平和血小板聚集功能的影响。结果发现：50mg／日、100mg／日均可使患者的血浆血栓素水平显著下降；三种剂量的阿斯匹林对血小板聚集功能都有显著的抑制作用，但以50mg／日、100mg／日效果为佳。据此我们推茬50mg／日作为防治糖尿病性视网膜病变的最佳药物剂量。     

Sight-threatening Diabetic Retinopathy at Presentation to Screening Services in Fiji

Purpose: To report the spectrum of retinopathy at first presentation to photoscreening services, to determine the proportion of patients that present with sight-threatening diabetic retinopathy (STDR), and to raise awareness of the burden of diabetic eye disease in Fiji.

Methods: This retrospective observational cohort study used data from the initial visit of all new patients presenting to the diabetes retinal screening service at the Pacific Eye Institute in Fiji over the 3-month period between July and September 2012. Patients were assessed using a detailed questionnaire regarding diabetes type, duration of disease, medications, complications and co-morbidities, and blood sugar control. Patients subsequently underwent non-mydriatic fundus photography according to Pacific diabetes retinal screening guidelines. Images were graded at the time of acquisition, and data were entered onto a computerized database. For the purposes of this study, information regarding retinopathy grading, visual acuity and patient demographics was used.

Results: A total of 522 new patients were screened over the 3-month period. STDR was observed in 27% of patients, with 15% observed to have bilateral STDR. Diabetes control was generally poor. Blindness and visual impairment were observed in 2.7% and 6.7% of the cohort, respectively.

Conclusion: Severe and advanced diabetic retinopathy was present in this population presenting to screening. This was observed 4 years after the formal expansion of the screening services and reflects the high prevalence of diabetes in the population. The need for increased public awareness and greater resource allocation into diabetes and its complications is emphasized.     

Ultrawide-field Fluorescein Angiography for Evaluation of Diabetic Retinopathy

Purpose

To investigate the advantages of ultrawide-field fluorescein angiography (FA) over the standard fundus examination in the evaluation of diabetic retinopathy (DR).

Methods

Ultrawide-field FAs were obtained in 118 eyes of 59 diabetic patients; 11 eyes with no DR, 71 eyes with nonproliferative diabetic retinopathy (NPDR), and 36 eyes with proliferative diabetic retinopathy (PDR), diagnosed by the standard method. The presence of peripheral abnormal lesions beyond the standard seven fields was examined.

Results

Ultrawide-field FA images demonstrated peripheral microaneurysms in six (54.5%) of 11 eyes with no DR and all eyes with moderate to severe NPDR and PDR. Peripheral retinal neovascularizations were detected in three (4.2%) of 71 eyes with NPDR and in 13 (36.1%) of 36 eyes with PDR. Peripheral vascular nonperfusion and vascular leakage were found in two-thirds of eyes with severe NPDR and PDR.

Conclusions

Ultrawide-field FA demonstrates peripheral lesions beyond standard fields, which can allow early detection and a close evaluation of DR.     

Genetics of Diabetic Retinopathy

Abstract

Multiple studies have shown that genetic factors may play an important role in determining an individual’s risk for the development of diabetic retinopathy (DR) and progression to proliferative DR. However, consistent and definitive genetic associations with DR across broad populations have been not been established. Numerous genes have been studied for their association with DR and the results of these investigations have most specifically pointed to three specific genes that are likely involved in DR development and progression. The gene coding for vascular endothelial growth factor, aldose reductase, and the receptor for advanced glycation end products have been extensively evaluated, and specific polymorphisms of these genes have been suggested to potentially increase the risk of DR development. In this paper, we have reviewed the published literature on the genetics of DR and the potential implications for DR development and progression.     

福辛普利、转化生长因子β_1与糖尿病视网膜病变

目的探讨血清转化生长因子β1(TGFβ1)在糖尿病视网膜病变(DR)发病中的意义以及血管紧张素转化酶抑制剂(ACET)福辛普利对DR患者的治疗效果及对DR患者血清TGFβ1水平的影响。方法将80例2型糖尿病(2-DM)患者及20例健康人群分为5组,采用双抗体夹心酶联免疫吸附法(ELISA)检测其血清TGFβ1水平并进行比较。将45例背景型DR患者分为2组,25例福辛普利治疗组给予福辛普利治疗6个月,行眼底荧光造影(FFA)检查观察患者治疗前后眼底病变的变化并与药物对照组比较,同时检测患者治疗前后血清TGFβ1水平的改变。结果糖尿病组与正常对照组比较,血清TGFβ1水平升高,差异有显著性意义(P<0．01)。糖尿病三组之间比较,PDR组血清TGFβ1水平高于背景型DR组;背景型DR组血清TGFβ1水平高于NODR组,三组之间差异均有显著性意义(P<0．05)。苯那普利治疗后背景型DR患者血清TGFβ1水平及UAER降低,与治疗前比较,差异均有显著性意义(P<0．05)。福辛普利治疗前后DR眼底的变化与药物对照组比较,无显著性差异(P>0．05)。结论DR患者血清TGFβ1水平显著升高,且随病变进展进一步升高,TGFβ1可能在DR发病及其进展中发挥重要作用。福辛普利通过抑制背景型DR患者产生TGFβ1,从而发挥对视网膜可能的保护用,但短期内观察眼底的病变无明显改善。     

A Study of VEGF Gene Polymorphism in Egyptian Patients with Diabetic Retinopathy

Background: There are subgroups of patients with diabetes mellitus (DM) in whom diabetic retinopathy (DR) does not develop despite poor long-term control of their disease, while others exercising fairly good control, develop retinopathy. So, we aimed to investigate the association of DR with ?2578 polymorphism of the vascular endothelial growth factor (VEGF) gene, which has been reported to be associated with increased VEGF production, in Egyptian diabetic patients.

Materials and Methods: This is a case control study in which 148 diabetic patients were enrolled. Among them, 44 subjects had proliferative diabetic retinopathy (PDR), 30 had non-proliferative diabetic retinopathy (NPDR), and 74 individuals without retinopathy served as controls. A single nucleotide polymorphism (SNP) of the VEGF gene, a C→A transversion at ?2578 (the C/A polymorphism), was investigated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

Results: We found a higher frequency of the polymorphic genotype in both the NPDR (66.7%) and PDR (72.7%) groups compared to the wild C/C genotype (33.3% in NPDR and 27.3% in PDR), but with no statistically significant difference from the control group. Significant association of the progression of DR to the polymorphic genotype was achieved at diabetes duration more than 20 years.

Conclusion: Despite of the higher frequency of both the polymorphic genotype and the A allele in cases with DR compared to the control group, there might be no significant association between the VEGF gene polymorphism and DR per se, unless it is longstanding.     

Utility Values Associated with Diabetic Retinopathy in Chennai,India

Abstract

Purpose: The aim of this study was to estimate utility values associated with different severity stages of diabetic retinopathy (DR) in India by a direct elicitation method (time-trade off, TTO) and indirectly by questionnaire.

Methods: People with diabetes aged 40 years and over were recruited from an on-going DR epidemiology study and a laser clinic in Chennai, India. Utility values were elicited using the direct TTO method and indirectly through a validated questionnaire (EQ-5D).

Results: Of 249 participants, 30 had no DR, 73 had non-proliferative DR, 114 had sight-threatening DR, and 32 were blind from DR (bilateral visual acuity <6/60). The mean TTO utility value was 0.73 (standard deviation, SD, 0.31). TTO utility values decreased with increasing severity of DR (p?<?0.001) and were significantly lower among participants with sight threatening DR (0.70, SD 0.33) and blindness (0.55, SD 0.24) compared to those with no DR (0.89, SD 0.25) after adjustment for sociodemographic and clinical factors. Blindness from DR was independently associated with a lower EQ-5D utility value. The utility value derived from EQ-5D (0.06) associated with being blind from DR was substantially lower than that of the TTO utility value (0.55).

Conclusions: This study provides estimates of utility values that can be used in economic evaluations of DR screening strategies in India. The relatively low utility values associated with blindness highlights the importance of screening programs for early detection of the sight-threatening stages to prevent vision loss from DR in this setting.