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.     

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.     

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.     

外周血循环内皮细胞、血浆内皮素与糖尿病视网膜病变的关系

目的 探讨血管内皮细胞(vascular endothelial cells,VEC)损伤及损坏后的功能变化在糖尿病视网膜病变(diabetic retinopathy,DR)发病机制中的作用。 方法 检测55例糖尿病(diabetes mellitus,DM)患者(包括无视网膜病变组20例,背景型视网膜病变组20例,增生型视网膜病变组15例)及正常人外周血循环内皮细胞(circulating endothelial cells ,CEC)数及血浆内皮素 (endothelin,ET)水平变化,并进行比较。 结果 DM患者CEC数及ET水平显著高于正常人(P<0.001),两指标呈显著正相关(r=0.738,P<0.001,n=55)。CEC数及ET水平随DR程度加重而增高。 结论 VEC损伤及由此所致的血浆ET水平增高,可能共同参与了DR的病理过程。(中华眼底病杂志,2000,16:166-168)