增生性糖尿病视网膜病变玻璃体中结缔组织生长因子的定量分析

目的 定量测定结缔组织生长因子(connective tissue growth factor,CTGF)在增生性糖尿病视网膜病变(proliferative di-abetic retinopathy,PDR)患者玻璃体中的浓度,探讨其在PDR发病机制中的作用.方法 采用双抗体夹心酶联免疫吸附测定法定量检测27眼PDR、5眼非增生性糖尿病视网膜病变及5眼正常对照组玻璃体中CTGF的浓度.结果 PDR组玻璃体中CTGF浓度(567.09±181.15)ng·L-1明显大于对照组(128.06±57.28)ng·L-1及非增生性糖尿病视网膜病变组(150.70±52.39)ng·L-1(P均<0.01).PDR组中Ⅵ期患者玻璃体中CTGF浓度(706.17±88.78)ng·L-1大于Ⅳ期(349.20±110.60)ng·L-1及Ⅴ期(526.70±122.50)ng·L-1(P均<0.01).结论 PDR患者玻璃体中CTGF浓度升高可能与视网膜新生血管纤维膜的形成有关,CTGF在PDR发展过程中起着一定作用.     

miR-106调控CC趋化因子配体2对增生型糖尿病视网膜病变中人视网膜微血管内皮细胞增殖、血管生成和炎症反应的影响

目的　探讨miR-106调控CC趋化因子配体2（CCL2）对增生型糖尿病视网膜病变（PDR）中人视网膜微血管内皮细胞（HRMEC）增殖、血管生成、炎症反应的影响。方法　GEO数据库筛选PDR中差异表达的miRNAs。高糖（25.0 mmol·L^-1葡萄糖,HG）诱导HRMEC建立PDR细胞模型,qRT-PCR检测miR-106和CCL2在正常葡萄糖（5.5 mmol·L^-1葡萄糖,NG）和HG培养条件下HRMEC中的表达。将PDR患者的血清外泌体做miR-106过表达处理后与HG处理的HRMEC共培养,同时干预HRMEC中CCL2的表达以探讨血清外泌体miR-106和CCL2在PDR中的功能。采用MTT法、小管形成实验和ELISA法分别检测各组HRMEC增殖、血管生成和炎症因子的表达。双荧光素酶报告实验用以验证miR-106和CCL2的靶向关系。结果　与NG组细胞中miR-106表达（1.04±0.10）、CCL2表达（1.02±0.09）相比,HG培养的HRMEC中miR-106表达（0.68±0.06）降低,CCL2表达（1.38±0.11）升高（均为P<0.05）。PDR患者血清外泌体中miR-106表达较NDR患者血清外泌体中表达降低（P<0.05）。与HG+PDR-exo+miR-NC组相比,HG+PDR-exo+miR-106 mimic组HRMEC活力降低,血管生成被抑制,细胞上清液中TNF-α、IL-1β和IL-6水平降低,而SOD、CAT和GSH水平升高（均为P<0.05）。双荧光素酶报告实验证实了CCL2是miR-106的一个靶点。与HG+PDR-exo+miR-106 mimic+oe-NC组相比,HG+PDR-exo+miR-106 mimic+oe-CCL2组HRMEC活力提高,血管生成被诱导,TNF-α、IL-1β和IL-6水平升高,而SOD、CAT和GSH水平降低（均为P<0.05）。结论　血清外泌体miR-106能够抑制CCL2表达,进而对PDR中的HRMEC增殖、血管生成和炎症反应起抑制作用。     

睫状体平坦部四切口玻璃体手术治疗严重增生性糖尿病视网膜病变

目的探讨睫状体平坦部四切口玻璃体手术治疗有广泛纤维血管膜增生的糖尿病视网膜病变（PDR）的临床效果。方法为病例对照试验。回顾性选择27例（28只眼）有广泛纤维血管膜增生的PDR Ⅵ期患者作为试验组,采用睫状体平坦部四切口玻璃体手术,双手进行眼内操作,如膜分离与切除,视网膜复位,眼内光凝硅油充填。选择同期有广泛纤维血管膜增生的PDR Ⅵ期患者30例（30只眼）作为对照组,由同一术者完成睫状体平坦部三切口玻璃体手术。结果试验组28只眼均顺利完成膜分离与切除,1只眼出现2个医源性视网膜裂孔。随访7～54个月,术后视网膜均复位,多数患者视力有不同程度提高。对照组2只眼有部分膜残留,3只眼出现4个医源性视网膜裂孔,随访12个月视网膜均复位,3只眼发生新生血管性青光眼。结论四切口玻璃体手术采用双手操作眼内剥膜,可明显提高手术效率,减少组织损伤,是治疗有广泛纤维血管膜增生的严重PDR的较好方法。（中华眼科杂志,2008,44：17—19）     

增生性玻璃体视网膜疾病眼内液血管内皮生长因子的表达

目的 观察增生性玻璃体视网膜疾病患者眼内液（房水、玻璃体）中血管内皮生长因子（VEGF）含量的变化规律，探讨VEGF在增生性玻璃体视网膜疾病发展变化中的作用。 方法 采用双抗体夹心酶联免疫吸附试验(ELISA)定量检测增生性玻璃体视网膜病变（PVR）、视网膜静脉阻塞（RVO）、增生性糖尿病视网膜病变（PDR）、新生血管性青光眼（NVG）患者组及正常对照组房水、玻璃体VEGF含量。 结果 PVR、RVO、PDR、NVG患者组房水及玻璃体VEGF含量均高于正常对照组，其差异均有统计学意义(P<0.05)；NVG、PDR、RVO、PVR患者组房水及玻璃体VEGF含量依次降低 ，其差异有统计学意义(P<0.05)；PVR、RVO、PDR、NVG患者组和正常对照组玻璃体VEGF含量均高于房水VEGF含量，其差异有统计学意义(P<0.05)；PVR患者病史与房水、玻璃体VEGF含量呈负相关（r分别为－0.819、－0.823，P＜0.05）；RVO患者病史与房水、玻璃体VEGF含量呈正相关（r分别为0.913、0.929，P＜0.05）；PDR患者玻璃体积血时间与房水、玻璃体VEGF含量呈正相关（r分别为0.905、0.920，P＜0.05）。 结论 增生性玻璃体视网膜疾病患者房水及玻璃体VEGF含量明显增高，VEGF可能在增生性玻璃体视网膜疾病发展变化中起着重要的作用。 (中华眼底病杂志， 2006， 22：313-316）     

雷珠单抗辅助玻璃体视网膜手术治疗增生型糖尿病视网膜病变

目的　观察雷珠单抗辅助玻璃体视网膜手术（vitreoretinal surgery,VRS）治疗Ⅴ期增生型糖尿病视网膜病变（proliferative diabetic retinopathy,PDR）的疗效。方法　将PDR（Ⅴ期）患者随机分为治疗组（42例42眼）和对照组（42例42眼）,治疗组患者行VRS手术前3~5 d辅助应用雷珠单抗,对照组患者仅行VRS。观察两组患者术中出血率、手术时间、医源性裂孔及眼内填充物等术中情况和术后1个月、3个月、6个月最佳矫正视力、黄斑中心凹视网膜厚度及并发症等情况。结果　治疗组和对照组患者手术时间分别为（78.07±8.58）min和（127.79±12.21）min,术中患者出血率、医源性裂孔发生率、硅油填充率治疗组分别为9.52%、2.38%、33.33%,对照组分别为40.48%、16.67%、69.05%,两组比较差异均有统计学意义（均为P<0.05）。术后1个月、3个月、6个月,治疗组的有效率分别为90.48%、76.19%、57.14%,对照组的有效率分别为61.90%、52.38%、47.62%;治疗组和对照组患者的黄斑中心凹视网膜厚度分别为（255.55±13.80） μm、（247.19±13.48） μm、（276.69±20.78）μm和（292.88±20.50） μm、（271.26±25.96） μm、（282.45±18.70）μm。术后1个月、3个月治疗组患者的有效率、黄斑中心凹视网膜厚度及并发症的发生均明显低于对照组,差异均有统计学意义（均为P<0.05）。术后3个月治疗组患者的无菌性眼内炎症和视网膜再增殖发生率均低于对照组,差异均有统计学意义（均为P<0.05）;两组患者玻璃体再出血和新生血管再生长发生率差异均无统计学意义（均为P>0.05）。术后6个月治疗组患者的有效率、黄斑中心凹视网膜厚度及并发症发生率和对照组比较差异均无统计学意义（均为P> 0.05）。结论　雷珠单抗辅助VRS,能显著减少PDR患者术中的出血,缩短手术时间、降低医源性裂孔发生率和硅油填充率,有助于PDR患者术后近期视功能的恢复,提高了PDR患者的近期临床治疗效果。     

血管生成素样蛋白4(ANGPTL4)在增生型糖尿病视网膜病变患者血清和玻璃体中的表达

目的 定量检测增生型糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)患者血清和玻璃体内血管生成素样蛋白4(ANGPTL4)的表达,并探索ANGPTL4在PDR中的作用.方法 收集需经睫状体平坦部20G标准三通道玻璃体切割术的患者共69例69眼,其中,PDR组49例49眼,对照组(非糖尿病性全层黄斑裂孔或黄斑裂孔性视网膜脱离患者)20例20眼.酶联免疫吸附试验定量检测两组患者血清及玻璃体标本中ANGPTL4表达水平.使用SPSS 17.0统计学软件对数据进行统计学分析.结果 PDR组患者血清中ANGPTL4的表达水平(30.761±2.996)ng · mL-较对照组患者(35.912±1.763)ng·mL-低,差异有统计学意义(t=-8.851,P=0.000);玻璃体ANGPTL4的表达水平(14.723±1.324)ng·mL-1较对照组患者(7.892±0.812)ng·mL-1高,差异有统计学意义(t =24.642,P=0.000).两组患者血清中ANGPTL4表达水平均高于其玻璃体,且PDR组患者血清与玻璃体中ANGPTL4变化呈正相关关系(r=0.884,P=0.000);对照组患者血清与玻璃体中ANGPTL4变化亦呈正相关关系(r=0.881,P=0.000).PDR组患者血清及玻璃体中ANGPTL4表达水平均与甘油三酯呈正相关,与高密度脂蛋白呈负相关关系(均为P＜0.05),而与其他临床指标无明显相关关系.术前玻璃体内注射雷珠单抗与不注射PDR患者血清及玻璃体中ANGPTL4表达水平,差异均无统计学意义(均为P＜0.05).结论 PDR患者血清中ANGPTL4表达水平低于非糖尿病患者,其可能在糖尿病患者糖脂代谢的病理生理过程中发挥了一定的作用;PDR患者玻璃体中ANGPTL4表达水平高于非糖尿病患者,其可能参与了PDR的发生发展过程,针对PDR患者玻璃体中ANGPTL4的高表达进行干预可能为未来PDR患者的治疗提供新的思路.     

Avastin在增生型糖尿病视网膜病变手术中的应用及作用机制

背景研究表明avastin辅助玻璃体手术治疗严重增生型糖尿病视网膜病变（PDR）能减少手术并发症,降低手术难度,可能与avastin抑制新生血管的形成有关,但缺乏组织病理学证据。目的探讨玻璃体腔注射avastin辅助的玻璃体手术治疗严重PDR减少手术并发症的可能机制。方法病例对照研究。收集严重PDR患者24例24眼,按照手术方式将患者分为单纯手术组10例10眼,单纯进行玻璃体切割术;avastin＋手术组14例14眼,行玻璃体手术前2周一次性玻璃体腔注射25g/L avastin0．06ml。收集术中剥离的视网膜前膜行苏木精一伊红染色,观察组织病理学改变。应用免疫组织化学染色法检测CD34在新生血管内皮细胞中的表达,比较2组患者视网膜前膜新生血管的密度及单细胞新生血管密度。结果单纯手术组与avastin＋手术组患者的人口基线特征和眼部基线特衙比较差异均无统计学意义（P〉0．05）。单纯手术组视网膜＋＋＋级新生血管者10眼,avastin＋手术组为1眼,二者比较差异有统计学意义（P〈0．01）。组织病理学结果表明,单纯手术组增生膜内可见较多散在的毛细血管型新生血管,多数由单个血管内皮细胞构成管腔,部分区域可见较多出血;avastin＋手术组增生膜玻璃样变的成分增多,新生的毛细血管明显减少或消失。免疫组织化学检测证实,新生血管内皮细胞中CD34均呈强阳性表达,avastin＋手术组400倍镜下每个视野新生血管密度为（15．40±7．42）个,单纯手术组为（8．00±3．80）个,差异有统计学意义（Z=-4．102,P〈0．01）;avastin＋手术组单细胞新生血管密度为（1．88±1．71）个,单纯手术组为（0．45±0．56）个,差异有统计学意义（Z=-4．137,P〈0．01）。结论Avastin可抑制增生膜新生血管芽的形成,阻断新生血管的发生,减轻视网膜水肿,这可能是其辅助玻璃体手术治疗晚期PDR、降低手术难度的主要机制。     

肝细胞生长因子受体在增生性糖尿病视网膜病变视网膜前膜中的表达

目的：肝细胞生长因子（hepatocyte growth factor receptor,HGF)是具有多种生物活性的细胞因子,通过与受体结合转导生物学效应,发挥促细胞增生,促运动和促形态发生等作用。本实验目的在于观察增生性糖尿病视网膜病变（poliferative diabetic retinopathy,PDR)不同时期视网膜前膜（epiretinal memebranes,ERM)中HGF受体的表达情况。方法：13例PDR患行玻璃体切除术剥离视网膜前膜,采用免疫组化染色方法观察HGFR在ERM中的表达情况。结果：在7例IV,V期ERM标本中,5例HGF受体阳性表达,阳性表达率为71．4％;6例PDR VI期增殖前膜标本中,有5例HGF受体阳性表达,阳性表达率为83．3％,HGF受体阳性表达率与PDR分期无显性差异（P＜0．05）,结论：HGF受体在PDR视网膜前增生膜中表达,提示HGF可能参与了PDR增生膜的形成。     

SDF-1在增生性糖尿病视网膜病变中的表达及意义

目的 观察基质细胞衍生因子(stromal cell derivedfactor-1,SDF-1)在糖尿病视网膜新生血管膜和玻璃体中的表达水平,并探讨其与增生性糖尿病视网膜病变(proliferativedibetic retinopathy,PDR)程度的相关性.方法 采用免疫组织化学法,对12例PDR患者视网膜新生血管膜上的SDF-1表达情况进行检测.采用双夹心酶联免疫吸附测定法对PDR组20例、PDR引起的新生血管性视网膜病变组10例、增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)组20例和正常人组10例的玻璃体中SDF-1浓度进行检测.结果 在12例新生血管膜的标本中10例有SDF-1的阳性表达.PDR组玻璃体中,SDF-l浓度为(351.31±120.30)ng·L-1,PDR引起的新生血管性视网膜病变组为(1 161.49±103.43)ng·L-1,PVR组为(72.59±8.78)ng·L-1,正常人组<62.5 ng·L-1;PDR组与PDR引起的新生血管性视网膜病变组、PVR组SDF-1含量相比,差异均有统计学意义(t=-18.17、10.31,P<0.001).结论 SDF-1参与了糖尿病视网膜新生血管的形成;玻璃体中的SDF-1浓度与PDR程度呈正相关.     

肝细胞生长因子受体在增生型糖尿病视网膜病变视网膜前膜中的表达

目的观察增生型糖尿病视网膜病变(PDR)不同时期视网膜前膜(ERM)中肝细胞生长因子(HGF)受体的表达情况.方法13例PDR患者行玻璃体切割术剥离视网膜前膜.采用免疫组化染色方法观察HGFR在ERM中的表达情况.结果在7例Ⅳ、Ⅴ期ERM标本中,5例HGF受体阳性表达,阳性表达率为71.4%;6例PDR Ⅵ期增殖前膜标本中,有5例HGF受体阳性表达,阳性表达率为83.3%.HGF受体阳性表达率与PDR分期无明显关系(P＞0.05).结论HGF受体在PDR视网膜前增生膜中表达,提示HGF可能参与了PDR增生膜的形成.     

瘦素在增殖性糖尿病性视网膜病变和增殖性玻璃体视网膜病变中的表达

目的 通过检测瘦素在增殖性糖尿病性视网膜病变（proliferative diabetic retinopathy,PDR）和增殖性玻璃体视网膜病变（proliferative vitreous retinopathy,PVR）中的表达,探讨瘦素在PDR、PVR发生、发展过程中可能的调节机制。方法 分别用免疫组织化学染色的方法和酶联免疫吸附实验检测30例PDR患者、20例PVR病变患者眼内视网膜前膜中瘦素的表达,以及患者的血清、眼前房水、玻璃体液中瘦素的浓度。用Chi－Square Tests统计学方法分析和比较PDR、PVR与对照组之间瘦素表达的差异。结果 免疫组织化学染色结果：30例PDR患者中,有18例患者眼内视网膜前膜的瘦素受体呈阳性表达,阳性率为60%,与对照组比较,差异有统计学意义;20例PVR患者中,有3例患者眼内视网膜前膜的瘦素受体呈阳性表达,其中2例为血管纤维性视网膜前膜,1例为细胞纤维性视网膜前膜,阳性率为15%,与对照组比较,差异无统计学意义。ELISA结果：检测30例PDR患者的血清、眼前房水、玻璃体液中瘦素的浓度,与对照组之间差异有统计学意义（P＜0.05）;检测20例PVR患者的血清、眼前房水、玻璃体液中瘦素的浓度,与对照组之间差异无统计学意义（P＞0.05）。结论 瘦素可能主要是通过促进新生血管的生成参与到增殖性糖尿病性视网膜病变的发生、发展中,与增殖性玻璃体视网膜病变的发生、发展无明显相关性。     

VEGF在PDR中的表达及作用的研究

目的:研究增殖性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)患者血液、房水、玻璃体中血管内皮生长因子(vascular endothelial growth factor,VEGF)含量的变化,探讨VEGF与PDR的关系,为抗VEGF药物治疗的给药途径及剂量等提供理论依据。方法:采用双抗体夹心酶联免疫吸附测定法定量检测无糖尿病视网膜病变(NDR)组,单纯性糖尿病视网膜病变(BDR)组,增殖性糖尿病视网膜病变(PDR)组患者和正常对照组血浆中VEGF含量,还检测PDR患者房水、玻璃体中和正常对照组房水、玻璃体中VEGF含量,并进行综合分析。试剂盒购自美国R&D公司,其质量和灵敏度相对较高。结果:PDR组房水中VEGF含量有增高趋势,但与正常对照组比较,无统计学差异(P>0.05)。PDR患者玻璃体中VEGF含量明显增高,与正常对照组比较差异非常显著(P<0.01)。PDR组自身血浆、房水、玻璃体中VEGF含量比较有逐渐增高趋势,三者之间有显著性差异(P<0.01)。正常对照组血浆、房水、玻璃体中VEGF含量三者之间无显著性差异(P>0.05)。血浆VEGF含量在正常对照组中最高,而玻璃体中VEGF含量在PDR患者中最高。结论:PDR患者眼内尤其是玻璃体中VEGF含量大幅度增高,可能对促进DR发展恶化起了关键性的作用。在正常人,VEGF更多地存在于血浆中发挥其生物学效应。在严重DR患者中,玻璃体中异常地出现大量VEGF,推测来自缺血缺氧的视网膜,并可能有向眼前段扩散的趋势。     

抗VEGF辅助PPV治疗增生性糖尿病视网膜病变的效果及作用机制分析

目的:探讨抗血管内皮生长因子(vascular endothelial growth factor,VEGF)辅助玻璃体切割术(pars plana vitrectomy,PPV)治疗增生性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)的效果及作用机制.方法:将92例92眼行PPV的PDR患者,根据术前有无玻璃体腔注射雷珠单抗(intravitreal ranibizumab,IVR)分为单纯PPV组(41例41眼)和联合治疗组(51例51眼),其中联合治疗组于PPV术前5~7 d进行IVR.比较两组手术时间、电凝次数、硅油填充率、术后并发症发生率、术后3 mo患眼BCVA及手术前后不同时间点房水和玻璃体VEGF、色素上皮衍生因子(pigment epithelium-derived factor,PEDF)含量.结果:联合治疗组手术时间短于单纯PPV组,电凝次数少于PPV组,硅油填充、医源性视网膜裂孔及玻璃体再积血的几率均低于单纯PPV组,差异均有统计学意义(P<0.05);联合治疗组PPV时房水及玻璃体VEGF、PEDF含量低于单纯PPV组,差异均有统计学意义(P<0.05);联合治疗组术后3 mo患眼BCVA好于单纯PPV组,差异有统计学意义(P<0.05).结论:IVR联合PPV治疗PDR可降低PPV围手术期VEGF、PEDF水平,减少术中电凝次数,降低术后医源性视网膜裂孔及玻璃体再积血发生率,提高视力水平.     

Upregulation of RAGE and its ligands in proliferative retinal disease

We sought to study the presence of the receptor for advanced glycation endproducts (RAGE) and its ligands, advanced glycation endproducts (AGEs), S100/calgranulins and amphoterin (high mobility group box 1 protein; HMGB1), in the vitreous cavity and epiretinal membranes (ERMs) of eyes of patients with proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR). Undiluted vitreous specimens were collected from 30 eyes of 30 patients undergoing pars plana vitrectomy for repair of retinal detachment (RD) secondary to PDR (n = 15) or PVR (n = 15). The vitreous samples obtained from 10 eyes undergoing macular hole repair were used as controls. Epiretinal membranes were obtained from eight eyes with PDR and from 10 eyes with PVR. The levels of AGEs in the vitreous were measured using ELISA. The vitreous levels of soluble RAGE (sRAGE), S100/calgranulins and amphoterin were measured using Western blot analyses. The localization of RAGE and its ligands in ERMs was determined with immunohistochemistry. The vitreous levels of sRAGE were significantly increased in both PDR and PVR (p < or = 0.05) compared to control vitreous. In both PDR and PVR, the vitreous levels of AGEs (p < or = 0.01), S100/calgranulins (p < or = 0.05), and amphoterin (p < or = 0.01) were also elevated compared to control eyes. Expression of RAGE was detected in six of eight ERMs from eyes with PDR and eight of 10 ERMs from eyes with PVR. Many cells expressing RAGE also expressed vimentin, suggesting a glial cell origin. Ligands for RAGE were also detected in ERMs, with AGEs detected in five eyes with PDR and eight eyes with PVR. Similarly, S100 and amphoterin ERM expression was observed in six eyes with PDR; these ligands were also expressed in ERMs from eyes with PVR (8 and 7 cases, respectively). We conclude that RAGE and its ligands are increased in the vitreous cavity of eyes with PDR and PVR and are present in ERMs of eyes with these proliferative retinal disorders. These findings suggest a role for the proinflammatory RAGE axis in the pathogenesis of proliferative retinal diseases.     

Osteopontin expression in vitreous and proliferative retinal membranes of patients with proliferative vitreous retinopathy

AIM: To analyze osteopontin (OPN) expression in vitreous and proliferative retinal membranes of patients with proliferative vitreous retinopathy (PVR). METHODS: A total of 54 vitreous fluid samples were obtained between 2009 and 2010, which contained 45 with PVR (group A) and 9 without PVR (group B). Enzyme-linked immunosorbent assay was applied to quantify the OPN concentrations in vitreous fluid. Four samples of proliferative retinal membrane were also obtained at the time of vitrectomy, and their contents of OPN were measured by Real-time RT-PCR. RESULTS: The OPN levels in the vitreous fluid were 778.48±62.06ng/mL in group A and 452.99±32.52ng/mL in group B. The vitreous OPN levels in group A were significantly higher than those in group B and to rise by time in the early stages of PVR. The average OPN levels in the proliferative retinal membranes (F=0.14) were also higher than those in the retinal pigment cells (F=0) using Real-time RT-PCR. CONCLUSION: The high vitreous and proliferative retinal membrane OPN levels in PVR suggest that OPN might promote the development of PVR. The vitreous OPN concentrations are rising by the time in the early phases of PVR.     

Interleukin-8, nitric oxide and glutathione status in proliferative vitreoretinopathy and proliferative diabetic retinopathy

PURPOSE: To evaluate interleukin-8 (IL-8), nitric oxide (NO) and glutathione (GSH) profiles in vitreous humor and blood samples in patients with proliferative diabetic retinopathy (PDR) and in patients with proliferative vitreoretinopathy (PVR) and to compare the levels with those of controls. PATIENTS AND METHODS: NO concentrations were determined by using the Greiss reaction in plasma and vitreous humor samples. GSH levels were determined in both blood and vitreous humor samples, using DTNB, a disulfide chromogen. Vitreous IL-8 were assayed by ELISA. Twenty-three patients with PDR, 18 patients with PVR and 21 cadavers as the control group were included in the study. RESULTS: Plasma and vitreous NO levels were found to be 25.6 +/- 2.1 and 36.9 +/- 3.0 micromol/l in patients with PDR, 27.0 +/- 4.7 and 34.3 +/- 2.9 micromol/l in patients with PVR and 17.4 +/- 2.7 and 15.9 +/- 1.4 micromol/l in controls, respectively. Vitreous humor and plasma NO levels did not show any statistically significant difference between PDR and PVR groups. However, the values for vitreous in both groups were significantly higher than those of controls (p < 0.0001). Although IL-8 levels in vitreous samples of patients with PDR were not significantly different (79.6 +/- 9.7 pg/ml) from those of patients with PVR (42.2 +/- 7.3 pg/ml) (p = 0.06), the levels in both groups were significantly higher than those of controls (19.0 +/- 3.9 pg/ml) (p < 0.0001 and p < 0.05, respectively). Blood and vitreous GSH levels were found to be 5.3 +/- 0.4 micromol/g. Hb and 0.58 +/- 0.16 micromol/l in patients with PDR and 8.4 +/- 0.5 micromol/g. Hb and 15.7 +/- 2.2 micromol/l in patients with PVR and 12.0 +/- 1.1 micromol/g. Hb and 0.26 +/- 0.03 mmol/l in controls, respectively. Vitreous and blood GSH levels were significantly lower in patients with PDR compared to those with PVR (p < 0.0001 for both). CONCLUSION: Elevated levels of vitreous and plasma NO and vitreous IL-8 in PDR and PVR implicate a role for these parameters in the proliferation in these ocular disorders. GSH concentrations both in vitreous and blood samples of the PVR and PDR patients were much less than those observed in the control group. Lower GSH concentrations detected in PDR in comparison with those in PVR in vitreous humor and to a lesser degree in blood may play an important role in pathogenesis of new retinal vessel formation in patients with PDR. This also suggests that oxidative stress may be involved in the pathogenesis of PVR and particularly that of PDR.     

磷酸果糖激酶-2/果糖-2，6-二磷酸酶3在增生型糖尿病性视网膜病变患者玻璃体和血清中的表达及其相关性分析

目的　定量检测增生型糖尿病性视网膜病变（proliferative diabetic retinopathy,PDR）患者玻璃体和血清中磷酸果糖激酶-2/果糖-2,6-二磷酸酶3（phospofructokinase-2/fructose-2,6-bisphosphatase 3,PFKFB3）和血管内皮细胞生长因子（vascular endothelial growth factor,VEGF）的表达水平,探讨PFKFB3在PDR患者中可能的作用机制。方法　纳入确诊为PDR拟行单眼玻璃体切割术的患者42例（42眼）作为试验组,并根据术前是否行玻璃体内注射抗VEGF药物进一步将试验组分为非注药组和注药组,纳入同期诊断为全层黄斑裂孔（full thickness macular hole,FTMH）及黄斑前膜（preretinal membrane of the macula,PRMM）的20例（20眼）患者作为对照组。收集各组患者的玻璃体和血清标本,定量检测各组标本中的PFKFB3和VEGF表达水平,并进行统计学分析。结果　试验组患者玻璃体中PFKFB3水平为（463.17±381.28）ng·L^-1,明显高于对照组［（158.43±86.88）ng·L^-1］（t=4.919,P＜0.001）,试验组血清中PFKFB3水平为（153.45±83.78）ng·L^-1,明显高于对照组［（92.72±32.42）ng·L^-1］（t=4.098,P＜0.001）。非注药组患者玻璃体中PFKFB3水平为（797.29±387.44）ng·L^-1,明显高于注药组［（257.56±181.49）ng·L^-1］（t=5.230,P＜0.001）,而非注药组血清中PFKFB3水平与注药组差异无统计学意义（t=0.679,P=0.501）。非注药、注药组和对照组患者玻璃体与血清中的PFKFB3水平无相关性（非注药组:r=0.194,P=0.471;注药组:r=0.071,P=0.731;对照组:r=0.254,P=0.279）。试验组患者玻璃体中VEGF水平为（1713.50±1386.90）ng·L^-1,明显高于对照组［（205.52±92.93）ng·L^-1］（t=7.014,P＜0.001）;试验组血清中VEGF水平为（224.98±168.08）ng·L^-1,明显高于对照组［（86.80±36.51）ng·L^-1］（t=5.082,P＜0.001）。非注药组患者玻璃体中VEGF水平为（3399.72±510.06）ng·L^-1,明显高于注药组［（675.82±242.57）ng·L^-1］（t=20.014,P＜0.001）,非注药组血清中VEGF水平为（373.40±174.23）ng·L^-1,明显高于注药组［（133.65±73.10）ng·L^-1］（t=5.228,P＜0.001）。非注药、注药组和对照组患者玻璃体与血清中的VEGF水平均存在正相关关系（非注药组:r=0.952,P＜0.001;注药组:r=0.423,P=0.031;对照组:r=0.776,P＜0.001）。非注药组、注药组和对照组患者玻璃体中PFKFB3与VEGF水平均存在正相关关系（非注药组:r=0.865,P＜0.001;注药组:r=0.587,P=0.002;对照组:r=0.807,P＜0.001）,而在血清中仅注药组的PFKFB3与VEGF水平存在正相关关系（r=0.444,P=0.023）。结论　PFKFB3可能参与了PDR的发生发展过程;VEGF可能通过上调PFKFB3的表达而参与PDR的发生发展。