2型糖尿病患者糖尿病视网膜病变不同时期外周血内皮祖细胞的数量变化

目的 观察2型糖尿病患者糖尿病视网膜病变(DR)不同时期外周血内皮祖细胞(EPCs)的数量变化.方法 临床确诊为2型糖尿病的60例患者纳入研究.根据眼底及荧光素眼底血管造影(FFA)检查结果,将患者分为无视网膜病变(NDR)组、非增生型DR(NPDR)组及增生型DR(PDR)组,每组各20例.选择同期门诊体检正常者20名作为对照组.抽取所有受检者晨起空腹静脉血6 ml,采用流式细胞仪检测,并比较4组受检者外周血EPCs数量的变化情况.结果 对照组、NDR、NPDR及PDR组受检者外周血EPCs数量分别为(0.0179±0.0047)%、(0.0151±0.0086)%、(0.0123±0.1137)%、(0.0316±0.0294)%;PDR组受检者外周血EPCs数量较其他3组高,差异有统计学意义(x2=43.780,P＜0.05);NDR、NPDR组受检者外周血EPCs数量略低于对照组,差异无统计学意义(x2=5.244,P=0.73);NPDR组受检者外周血EPCs数量低于NDR组,但差异也无统计学意义(x2=6.016,P=0.12).结论 NDR、NPDR患者外周血EPCs数量降低,PDR患者外周血EPCs数量显著增高.

Abstract：

Objective To observe the amount of endothelial progenitor cells(EPCs)at different stages of diabetic retinopathy(DR)in patients with type 2 diabetes mellitus(DM).Methods Sixty patients with type 2 DM were divided into no DR(NDR)group,non-proliferative DR(NPDR)group and proliferative DR(PDR)group according to the examination of fundus and fundus fluorescein angiography,20patients in each group.Twenty healthy people were collected as the control group.6 ml blood samples were taken from all the subjects,and then the EPCs contents in peripheral blood were detected by flow cytometry.Results The EPCs contents in peripheral blood of the control,NDR,NPDR and PDR group were(0.0179±0.0047)% ,(0.0151±0.0086)% ,(0.0123±0.1137)% ,(0.0316±0.0294)% .The EPCs contents in peripheral blood of the PDR group was significantly higher than those in others(χ2=43.780,P＜0.05);the EPCs contents in peripheral blood of the NDR and NPDR group were slightly lower than that in the control group(χ2=5.244,P=0.73);the EPCs contents in peripheral blood of the NPDR group was lower than that in the NDR group(χ2=6.0 1 6,P=0.12).Conclusion The EPCs contents in peripheral blood decreases in NDR,NPDR patients,while significantly increases in PDR patients.     

2型糖尿病患者糖尿病视网膜病变不同时期外周血内皮祖细胞的数量变化

Objective To observe the amount of endothelial progenitor cells(EPCs)at different stages of diabetic retinopathy(DR)in patients with type 2 diabetes mellitus(DM).Methods Sixty patients with type 2 DM were divided into no DR(NDR)group,non-proliferative DR(NPDR)group and proliferative DR(PDR)group according to the examination of fundus and fundus fluorescein angiography,20patients in each group.Twenty healthy people were collected as the control group.6 ml blood samples were taken from all the subjects,and then the EPCs contents in peripheral blood were detected by flow cytometry.Results The EPCs contents in peripheral blood of the control,NDR,NPDR and PDR group were(0.0179±0.0047)% ,(0.0151±0.0086)% ,(0.0123±0.1137)% ,(0.0316±0.0294)% .The EPCs contents in peripheral blood of the PDR group was significantly higher than those in others(χ2=43.780,P＜0.05);the EPCs contents in peripheral blood of the NDR and NPDR group were slightly lower than that in the control group(χ2=5.244,P=0.73);the EPCs contents in peripheral blood of the NPDR group was lower than that in the NDR group(χ2=6.0 1 6,P=0.12).Conclusion The EPCs contents in peripheral blood decreases in NDR,NPDR patients,while significantly increases in PDR patients.     

糖尿病视网膜病变患者外周血内皮祖细胞数量及功能的变化

目的 观察糖尿病视网膜病变(DR)患者外周血内皮祖细胞(EPC)数量及功能变化.方法 选取DR患者12例(DR组)、糖尿病(DM)患者18例(DM组)及无DM的老年性白内障患者15例(对照组)纳入研究.密度梯度离心法收集各组患者外周血单个核细胞,培养后第10天采用流式细胞仪计数EPC细胞阳性率.采用二苯基四氮唑嗅盐(MTT)比色法、黏附能力测定实验及Transwell实验检测EPC的增生、黏附和迁移能力.结果 流式细胞仪检测结果显示,对照组、DM组及DR组培养后第10天EPC细胞阳性率分别为(45.190±1.287)％、(30.130±3.245)％、(37.370±2.501)％;3组间差异有统计学意义(F=27.690,P=0.001).MTT比色法检测结果显示,对照组、DM组及DR组吸光度[A,旧称光密度(OD)]值分别为0.330±0.047、0.225±0.042、0.120±0.029;3组间差异有统计学意义(F=29.327,P=0.000).黏附能力测定实验结果显示,对照组、DM组及DR组EPC细胞数分别为76.400±7.503、51.167±6.646、26.500±7.853;3组间差异有统计学意义(F=56.612,P=0.000).Transwell实验结果显示,对照组、DM组及DR组EPC细胞数分别为23.600±6.504、20.833±4.491、12.000±2.944;3组间差异有统计学意义(F=6.477,P=0.012).结论 DR患者外周血EPC数量减少,且其增生、黏附和迁移能力受损.     

增生型糖尿病视网膜病变患者外周血内皮祖细胞的数量变化及意义

目的 观察增生型糖尿病视网膜病变(PDR)患者外周血内皮祖细胞(EPCs)数量的变化规律,探讨EPCs在糖尿病视网膜病变(DR)发病机制中的作用.方法 PDR患者40例(PDR组)、单纯2型糖尿病患者30例(糖尿病组)及健康体检者20名(对照组)纳入研究.抽取3组受检者晨起空腹静脉血2 ml,反复离心过滤.采用流式细胞仪分析2×105个细胞,以CD34和(或)CD133双阳性细胞群为EPCs,观察并比较3组受检者外周血EPCs数量变化;同时分析PDR患者外周血EPCs数量与DR病程、糖化血红蛋白及血脂的相关性.结果 PDR组、糖尿病组及对照组受检者外周血EPCs计数分别为(49±12)、(35±11)、(90±25)个/ml,3组间差异有统计学意义(F=56.260,P＜0.05).相关性分析发现,PDR患者外周血EPCs数量与DR病程呈正相关(r=0.564,P＜0.05),与糖化血红蛋白(r=-0.170,P>0.05)及血脂(r=0.261,P>0.05)均无相关性.结论 PDR患者外周血EPCs数量较正常者明显降低;EPCs在DR发病机制中可能具有一定作用.

Abstract：

Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.     

增生型糖尿病视网膜病变患者外周血内皮祖细胞的数量变化及意义

Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.     

增生型糖尿病视网膜病变患者外周血内皮祖细胞的数量变化及意义

Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.     

糖尿病视网膜病变对大鼠循环外周血内皮祖细胞数量的影响

目的 观察糖尿病视网膜病变(DR)对循环外周血内皮祖细胞(EPCs)的影响.方法 雄性成年Wistar大鼠60只,随机分为正常对照组和糖尿病组.后者通过腹腔注射链脲佐菌素(STZ)建立DR模型.采用流式细胞仪分别计数建模后1周、1、3、6个月各组大鼠循环外周血EPCs数量,同时于各相应时间点摘除大鼠眼球行苏木精一伊红(...     

糖尿病视网膜病变对大鼠循环外周血内皮祖细胞数量的影响

Objective To observe the effect of diabetic retinopathy on endothelial progenitor cells (EPCs)from peripheral blood.Methods Sixty male Wistar rats were divided into control group and diabetes group.The rats in diabetes group were induced with streptozotocin(STZ)injection for diabetic retinopathy model.Flow cytometry was used to identify and count the number of EPCs from peripheral blood at 1 week.1,3 and 6 months after injection.All eyeballs were examined by hematoxylin and eosin (HE)staining,periodic acid-Schiffs(PAS)staining of trypsin-digested retinal vessels flat preparation and transmission electron microscope.EPCs count,and the relationship between DR morphological changes and EPCs count were compared and analyzed.Results The quantity of EPCs from peripheral blood at 1 week,1,3 and 6 months after STZ injection were 25±7,28±8,39±7,43±7 cells per 200 000 monocytes respectively,which decreased compared with the control group 45±4 cells per 200 000 monocytes(F=8.933,P＜0.0 1).The quantity of EPCs was gradually increased at 1 week,1,3 and 6 months after STZ injection,accompanied with responsive pathological changes of retinal structure and vessels.The thickness of retina at 1 week and 1 month after injection were reduced slightly.The number of retinal ganglion cells reduced,with the time passing by.Endothelial cells were edema,mitochondrial was swollen,capillary basement membrane was thicken,lumen was significant stenosis,lumen occlusion and retinal artery aneurysm were observed at 6 months after STZ injection.Conclusion The number of EPCs increases gradually throughout the development of DR.     

糖尿病视网膜病变对大鼠循环外周血内皮祖细胞数量的影响

Objective To observe the effect of diabetic retinopathy on endothelial progenitor cells (EPCs)from peripheral blood.Methods Sixty male Wistar rats were divided into control group and diabetes group.The rats in diabetes group were induced with streptozotocin(STZ)injection for diabetic retinopathy model.Flow cytometry was used to identify and count the number of EPCs from peripheral blood at 1 week.1,3 and 6 months after injection.All eyeballs were examined by hematoxylin and eosin (HE)staining,periodic acid-Schiffs(PAS)staining of trypsin-digested retinal vessels flat preparation and transmission electron microscope.EPCs count,and the relationship between DR morphological changes and EPCs count were compared and analyzed.Results The quantity of EPCs from peripheral blood at 1 week,1,3 and 6 months after STZ injection were 25±7,28±8,39±7,43±7 cells per 200 000 monocytes respectively,which decreased compared with the control group 45±4 cells per 200 000 monocytes(F=8.933,P＜0.0 1).The quantity of EPCs was gradually increased at 1 week,1,3 and 6 months after STZ injection,accompanied with responsive pathological changes of retinal structure and vessels.The thickness of retina at 1 week and 1 month after injection were reduced slightly.The number of retinal ganglion cells reduced,with the time passing by.Endothelial cells were edema,mitochondrial was swollen,capillary basement membrane was thicken,lumen was significant stenosis,lumen occlusion and retinal artery aneurysm were observed at 6 months after STZ injection.Conclusion The number of EPCs increases gradually throughout the development of DR.     

辛伐他汀对糖尿病视网膜病变大鼠外周血内皮祖细胞数量和视网膜病变的影响

目的 探讨辛伐他汀对糖尿病视网膜病变(DR)大鼠外周血内皮祖细胞(EPCs)数量及视网膜病变的影响.方法 雄性成年Wistar大鼠80只,以随机数字表法分为正常对照组、模型对照组、安慰剂组、辛伐他汀组,每组各20只大鼠.采用腹腔注射链脲佐菌素(STZ)建立DR大鼠模型.正常对照组、模型对照组不给予任何干预;辛伐他汀组予辛伐他汀20 mg/kg灌胃,1次/d;安慰剂组给予等量蒸馏水灌胃,1次/d.分别于1、4及12周时取静脉血,采用流式细胞仪分别计数各组大鼠外周血EPCs数量的变化.于12周时处死大鼠,摘除眼球行苏木精-伊红(HE)染色,采用伊文思蓝(EB)定量检测血视网膜屏障的破坏程度,免疫组织化学法分析CD31在大鼠视网膜中的表达.实时定量逆转录聚合酶链反应检测内皮型一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)及血管生成素-1(Ang-1)在视网膜的表达.对比分析EPCs的数量改变与视网膜病理变化的相互关系.结果 与正常对照组大鼠相比,注射STZ后1、4、12周时安慰剂组大鼠外周血EPCs数均降低;注射STZ后1、4、12周时辛伐他汀组大鼠外周血EPCs数明显高于模型对照组和安慰剂组,差异均有统计学意义(t=4.967,5.648,6.688,6.042,7.392,7.454;P＜0.05);模型对照组和安慰剂组大鼠外周血EPCs数相比,差异无统计学意义(t=0.525,-0.249,-0.619;P＞0.05);正常对照组和辛伐他汀组大鼠外周血EPCs数相比,差异均有统计学意义(t=6.733,2.794,-5.535;P＜0.05).组织病理学检查显示,正常对照组大鼠视网膜各层结构清晰、排列整齐、形态正常;模型对照组和安慰剂组大鼠视网膜细胞排列紊乱,细胞核肿胀、体积增大,视网膜组织水肿;辛伐他汀组大鼠视网膜各层组织水肿减轻,细胞排列渐规则.模型对照组和辛伐他汀组大鼠视网膜平均EB渗漏量较正常对照组显著增加,辛伐他汀组较模型对照组明显减少,差异均有统计学意义(F=65.808,P＜0.05).正常对照组、模型对照组和安慰剂组中表达CD31的阳性细胞数显著低于辛伐他汀组,组间CD31阳性细胞数比较,差异有统计学意义(F=24.799,P＜0.05);eNOS在模型对照组表达较正常对照组明显减弱,辛伐他汀组表达强度较模型对照组增强(t=-2.750,2.230;P＜0.05);iNOS和Ang-1在模型对照组表达较正常对照组明显增强,辛伐他汀组表达强度较模型对照组减弱,差异均有统计学意义(t=3.881,-1.144,4.244,-1.458;P＜0.05).安慰剂组视网膜EB渗漏量、eNOS、iNOS及Ang-1的相对表达量与模型对照组相比,差异均无统计学意义(t=0.480,-0.877,0.062,0.220;P＞0.05).结论 辛伐他汀可动员DR大鼠外周血EPCs,诱导视网膜内皮细胞迁徙分化,减缓DR进展.其可能机制为调节eNOS和iNOS等内皮形成相关因子.     

2型糖尿病患者DR不同时期血浆钙卫蛋白水平的检测和比较

背景 糖尿病视网膜病变(DR)时视网膜血管的中性粒细胞浸润,造成血管内皮损伤.钙卫蛋白主要存在于中性粒细胞溶酶体外的细胞质中,通过促进中性粒细胞的聚集参与血管损伤的生物学行为,是一种潜在的临床炎症标志物.钙卫蛋白在DR进展过程中的作用尚不清楚. 目的 探讨钙卫蛋白在2型糖尿病患者DR不同时期的表达差异及其在DR发生发展过程中的可能作用. 方法 本研究为病例对照研究.临床确诊为2型糖尿病的患者60例纳入研究,根据检眼镜下眼底表现和荧光素眼底血管造影( FFA)检查结果将患者分为无DR组(20例)、非增生型糖尿病视网膜病变(NPDR)组(20例)和PDR组(20例),并纳入门诊体检的20例健康者为对照组,对照组与2型糖尿病组受检者的年龄、性别及血液生化检测指标结果相匹配.于清晨抽取受试者空腹静脉血2 ml,ELISA法测定各组受检者血浆钙卫蛋白的质量浓度. 结果 对照组、无DR组、NPDR组和PDR组受检者外周血浆中钙卫蛋白的质量浓度分别为(57.70±12.29)、(72.07±10.14)、(87.70±10.37)、(94.36±9.40)ng/L,4个组间的总体差异有统计学意义(F=73.09,P＜0.01).PDR组受检者外周血钙卫蛋白的质量浓度最高,与其他3个组间的两两比较差异均有统计学意义(q =20.157、10.648、4.497,P＜0.01);NPDR组受检者外周血钙卫蛋白的质量浓度高于无DR组,差异有统计学意义(q=6.216,P＜0.01).结论 血浆钙卫蛋白质量浓度的变化可能在2型糖尿病患者DR的发生发展过程中起一定作用.     

不同程度糖尿病视网膜病变患者血浆炎性因子检测

目的研究不同程度糖尿病视网膜病变(DR)患者血浆肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、可溶性白介素-2受体(IL-2R)及C反应蛋白(CRP)水平,为探究炎性反应在DR发病机制中的作用提供依据。设计前瞻性比较性病例系列。研究对象2型糖尿病但无DR患者21例(DM无DR组),非增生性DR患者17例(NPDR组),增生性DR患者23例(PDR组),无糖尿病患者23例作为对照组(无DM组)。方法记录患者年龄、性别、病史资料。行视力、眼压、裂隙灯及眼底检查。采集静脉血检测糖化血红蛋白(HbA1c)、空腹血糖(Glu)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL)、高密度脂蛋白胆固醇(HDL)、甘油三酯(TG)、同型半胱氨酸(Hcy)、肌酐(CR),以及炎症相关指标TNF-α、IL-2R、IL-8 IL-6及CRP的水平。主要指标血浆炎性因子浓度。结果四组之间年龄、性别、既往高血压、冠心病及脑梗塞病史、TC、LDL、HDL、TG、Hcy和血浆CR均无显著统计学差异。HbA1c在PDR组(7.51±2.03)%、NPDR组(7.48±1.49)%、DM无DR组(7.09±1.54)%,均显著高于无DM对照组(5.35±0.54)%(P<0.001)。TNF-α水平在PDR组及NPDR组(42.54±31.80及32.07±28.84 pg/ml)显著高于无DM组(10.30±5.35 pg/ml)及DM无DR组(12.63±6.65 pg/ml)(P<0.001)。IL-8水平在PDR组及NPDR组(157.26±200.16及197.45±331.08 pg/ml)显著高于无DM组(30.23±29.41 pg/ml)及DM无DR组(29.70±22.04 pg/ml)(P=0.006)。四组之间IL-2R、IL-6及CRP浓度均无显著统计学差异。结论NPDR及PDR患者血浆炎性因子TNF-α及IL-8显著高于DM无DR及无DM对照者。血浆TNF-α升高为PDR的重要危险因素。血浆炎性因子与肌酐水平显著正相关,全身高炎症状态可能是糖尿病肾病及DR的共同危险因素。