晚期糖基化终末产物对人血管内皮细胞增殖及凋亡的影响

目的研究晚期糖基化终末产物(AGEs)对人血管内皮细胞增殖及凋亡的影响,探讨糖尿病难愈创面新生血管化障碍或延迟的机理.方法不同作用时间及浓度AGE修饰的人血清白蛋白(AGE-HSA)与人血管内皮细胞(ECV304)在体外共同培养,用四唑盐(MTT)比色试验和细胞计数检测AGE-HSA对血管内皮细胞的增殖抑制作用,并用台盼蓝排斥试验检测细胞活力.采用FITC Annexin-V及碘化丙碇(PI)染色,用流式细胞仪检测凋亡细胞百分率,同时应用荧光共聚焦显微镜观察凋亡或死亡细胞FITC Annexin-V和PI的荧光染色,并用透射电镜和光镜观察凋亡细胞特异性的形态学改变.结果内皮细胞在12.5、25和50μg/ml AGE-HSA培养条件下,第2天细胞增殖数量与对照组比较无明显差异性,第4天和第6天则显著低于对照组(P＜0.01);而内皮细胞经100或200μg/ml AGE-HSA干预6h,MTT法测其OD值与对照组比较有明显差异(P＜0.01),同时内皮细胞出现特异性的凋亡形态学变化以及FITC Annexin-V阳性和/或PI阳性的细胞逐渐增多,且凋亡或死亡细胞数量随AGE-HSA作用时间及浓度的增加而增多.结论AGE-HSA能抑制内皮细胞增殖,并可以诱导其凋亡,而且与作用时间及浓度相关.提示AGEs可能是引起糖尿病难愈创面中新生血管化障碍或者延迟的机制之一.     

AGE与RAGE相互作用通过活性氧引起足细胞凋亡

目的：探讨晚期糖基化终末产物（AGE）对足细胞凋亡的影响,及氧化应激在其中的作用。方法：小鼠足细胞株由美国纽约西奈山医学院Peter Mundel教授馈赠。用钙磷脂结合蛋白Ⅴ-荧光异硫氰酸盐（FITC）和碘化物（PI）标记细胞,采用荧光激活细胞分类（FACS）法来计数凋亡和坏死的足细胞。Dharmacon On TargetPlus SMARTpool si RNA试剂和Amaxa RNAi nucleofection试剂盒成功转染si RNA到足细胞。绿荧光蛋白载体证明转染的有效性,分别采用Western Blot和实时定量PCR（RT-PCR）方法来检测si RNA转染足细胞后AGE受体蛋白（RAGE）靶基因蛋白质和mRNA的表达。用LS50B型荧光分光光度计测活性氧,根据波长485nm在530nm发射的荧光来判断活性氧（ROS）的产生。观察活性氧的清除剂N-乙酰基-半胱氨酸（NAC）能否减少AGE-BSA诱导的足细胞凋亡。结果：AGE引起足细胞的凋亡呈剂量依赖性,随AGE浓度的增大,凋亡的发生率逐渐升高;RAGE siRNA能减少60%~70%RAGE mRNA和蛋白质的表达;ROS的清除剂NAC可明显减少AGE-BSA引起的ROS的产生和足细胞凋亡。结论：AGE与RAGE作用后活性氧产生增加,活性氧的增加可能是AGE引起足细胞凋亡的途径之一,可通过抗氧化减少ROS的产生延缓糖尿病肾病的进展。     

晚期糖基化终末产物及受体在实验性2型糖尿病大鼠种植体周围的变化及表达

目的 建立2型糖尿病大鼠动物模型,探讨晚期糖基化终末产物及其受体在实验性2型糖尿病大鼠种植体骨整合过程中的变化及表达.方法 45只3个月龄SD健康雄性大鼠,将大鼠随机分为糖尿病模型组25只和正常对照组20只.首先建立2型糖尿病大鼠模型,建模成功后将模型大鼠随机分为DM组和DM种植组,每组10只.将20只正常组大鼠随机分为正常对照组和正常种植组,每组10只.分别于正常种植组和DM种植组的胫骨近骺端植入纯钛种植体,植入10周后于下腔静脉采血,保存所采集标本,用RF-5301PC型荧光分光光度计测定血清中AGEs含量的变化.硬组织标本采用不带种植体脱钙切片,以正常组为对照,HE染色后用免疫组织化学方法检测种植体周围RAGE的表达.结果 10周后,DM种植组和DM组与正常对照组和正常种植组相比,血清中AGEs的变化差异有统计学意义（P ＜0.05）,正常种植组和DM种植组与正常对照组的种植体周围骨组织RAGE表达比较,差异均有统计学意义（P ＜0.05）;DM种植组与DM组比较,差异亦有统计学意义（P ＜0.05）.结论 种植体骨组织愈合过程中AGEs和RAGE相互作用是影响2型糖尿病种植体骨结合的机制之一.     

尿毒清对晚期糖基化终末产物作用下肾小球系膜细胞影响

目的观察尿毒清对晚期糖基化终末产物（advanced glycosylation end products, AGEs）作用下肾小球系膜细胞的影响,探讨中药尿毒清在治疗糖尿病肾脏疾病（diabetic kidney dis-ease,DKD）中对肾小球系膜细胞的保护作用。方法采用冻干牛血清白蛋白和糖制备无内毒素的AGE-BSA及人工培养牛肾小球系膜细胞,培养时分为3组,在加入终浓度为0.25 mg/ml AGEs 的同时给予浓度为0.2 mg/ml的尿毒清成分溶液为尿毒清组,加入终浓度为0.25 mg/ml AGEs 为 AGEs组,同时设空白对照组,应用 MTT比色法,观察尿毒清组对体外 AGEs 培养的牛肾小球系膜细胞增殖的影响;以系膜细胞与 Alcain Blue（AB,阿利新蓝）结合量为指标,观察尿毒清组对 AGEs导致的系膜细胞膜表面电荷的影响;利用 AO/EB染料观察尿毒清组对 AGEs 诱导作用下的牛肾小球系膜细胞凋亡的影响,并利用荧光显微镜观察尿毒清的抑制作用。结果尿毒清组可以有效地抑制 AGEs对牛肾系膜细胞的促进增殖作用;对抗 AGEs 诱导的牛肾小球系膜细胞表面电荷的影响;减少 AGEs诱导的牛肾小球系膜细胞凋亡。结论尿毒清组能减少对晚期糖基化终末产物作用下肾小球系膜细胞增殖和凋亡,保护肾小球系膜细胞表面电荷。     

晚期糖基化终末产物与骨性关节炎相关性的研究进展

晚期糖基化终末产物(advanced glycation end products,AGEs)是衰老致骨性关节炎的重要因素,主要通过受体(receptors for advanced glycation end products,RAGE)途径,引起软骨基质降解与软骨细胞凋亡、促进软骨及滑膜组织的炎性反应等方式在OA发病中发挥重要作用。AGEs是OA领域的研究新热点,但其分子机制还未完全阐明,对AGEs的研究将有助于明确OA的发病机制,同时为OA的治疗策略提供了新的观念。     

血液透析患者可溶性晚期糖基化终末产物受体与颈动脉粥样硬化的关系

目的：探讨血液透析患者可溶性晚期糖基化终末产物受体（sRAGE）与颈动脉粥样硬化之间的关系。方法：检测76名血透患者的sRAGE水平以及颈动脉粥样硬化超声指标,并选取35名年龄、性别相匹配的健康对照,对sRAGE、颈动脉粥样硬化超声指标及相关临床资料进行分析。结果：血液透析患者的sRAGE水平显著高于健康对照组（P〈0.01）。有糖尿病、冠心病史的血透患者的sRAGE水平较无糖尿病、冠心病史者低（均P〈0.05）。多元线性回归分析显示,sRAGE与颈动脉内-中膜厚度（IMT）呈负相关（P〈0.05）。结论：血透患者sRAGE水平与颈动脉粥样硬化呈负相关,提示sRAGE是血透患者的一种血管保护因子。     

晚期糖基化终末产物与慢性肾功能衰竭

晚期糖基化终末产物 (AGE)是蛋白质与还原糖发生非酶促糖基化反应产物 ,AGE作为一种新的尿毒症毒素 ,与尿毒症及透析多种并发症或合并症有关。本文综述AGE在慢性肾功能衰竭中的致病作用及可能采取的防治措施。     

小檗碱在高糖及糖基化终末产物诱导足细胞损伤中的保护作用

目的探讨小檗碱对糖基化终末产物(AGE)和高糖诱导下足细胞损伤及其骨架蛋白的影响及机制。 方法以条件永生性人足细胞作为研究对象,于含10%胎牛血清及100 U/L γ-干扰素的RPMI 1640培养液中进行体外培养,细胞增殖并诱导分化后进行分组处理。分别用高糖(30 mmo/L)、AGE(100 μg/ml)、小檗碱(10 μmo/L)处理48 h后,激光共聚焦检测技术观察纤维状肌动蛋白(F-actin),球状肌动蛋白(G-actin)变化;原位细胞免疫组化检测cspase-3,nephrin表达。采用SPSS13.0统计软件包进行统计学分析。 结果共聚焦显微镜下观察显示,高糖及AGE作用下,足细胞F-actin出现重排,G-actin易位,小檗碱干预后有所恢复。免疫组化结果显示,对照组及高糖组几乎未见capase-3阳性表达,但高糖＋AGE组,capase-3呈阳性表达(F＝99.339,P<0.001);高糖＋AGE组nephrin表达显著降低(F＝165.84,P<0.001),与对照组及高糖组比较,差异均有统计学意义。小檗碱作用后,高糖＋AGE组capase-3的水平下降(F＝6.927,P＝0.048),nephrin表达水平升高(F＝165.84,P＝0.025),差异均有统计学意义。 结论在持续的高糖和AGE作用下,可引起足细胞骨架蛋白F-actin、G-actin重构及分布异常,并诱导足细胞凋亡,小檗碱能改善高糖和AGE引起的足细胞骨架蛋白损伤,并抑制足细胞的凋亡,其机制可能与nephrin的参与有关。     

晚期糖基化终末产物与慢性肾功能衰竭

晚期糖基化终末产物（AGE）是蛋白质与还原糖发生非酶促糖基化反应产物，AGE作为一种新的尿毒症毒素，与尿毒症及透析多种并发症或合并症有关，本文综述AGE在慢性肾功能衰竭中的致病作用及可能采取的防治措施。     

晚期非酶糖基化终末产物与糖尿病肾病

非酶糖基化终末产物（ＡＧＥｓ）可以造成组织细胞结构和功能的损害。ＡＧＥｓ可以引起肾脏细胞外基质的堆积，影响细胞代谢，诱发蛋白尿和肾小球肥大，在糖尿病肾病的发生和发展中起着重要作用。     

人类关节滑膜细胞表达晚期糖基化终产物受体

目的 进一步探讨晚期糖基化终产物修饰的β２－微球蛋白（ＡＧＥ－β２ｍ）对关节固有细胞的生物学作用,确定人类关节滑膜细胞是否表达对ＡＧＥ特异的受体。方法 分离、培养人关节Ａ型和Ｂ型滑膜细胞,用免疫组织化学法及流式细胞仪法分别观察滑膜细胞表面ＡＧＥ受体１（ＡＧＥ－Ｒ１）,ＡＧＥ受体２（ＡＧＥ－Ｒ２）、ＡＧＥ受体３（ＡＧＥ－Ｒ３）及ＡＧＥ受体（ＲＡＧＥ）的表达,用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术     

Increased receptor for advanced glycation end products in spermatozoa of diabetic men and its association with sperm nuclear DNA fragmentation

Although the majority of patients with diabetes have disorders in sexual function, associations between diabetes mellitus and sperm function at the molecular level are largely unknown. As receptor for advanced glycation end products plays a key role in many diabetic complications, we hypothesised that it may be involved in sperm nuclear DNA fragmentation. RAGE levels were determined using ELISA and western blot analysis in sperm samples from 32 diabetic and 35 nondiabetic men. Sperm DNA fragmentation was assessed using TUNEL assay. Diabetic men had significantly higher mean levels of RAGE protein (P < 0.001) and DNA fragmentation (P < 0.001) in spermatozoa. Sperm RAGE was directly correlated to sperm DNA fragmentation in diabetic men (r = 0.81, P < 0.001). The high positive correlation between RAGE levels and nuclear DNA fragmentation in spermatozoa of diabetic men suggests a central role of RAGE in disturbances in sexual function of diabetic men.     

胃癌组织中RAGE和配体HMGB1表达的临床研究

目的：分析胃癌及相应癌旁组织中的晚期糖基化终产物受体（receptor for advanced glycation end product,RAGE）及其配体高迁移率族蛋白B1 （high mobility group box-1,HMGB1）的表达情况,探讨RAGE、HMGB1表达与胃癌患者临床病理因素的关系以及两者对预后的影响.方法：应用免疫组织化学的方法检测1 09例胃癌组织和30例癌旁组织中RAGE和HMGB1的表达,运用美国Media Cybernetics公司Image-Pro？ Plus 6.2软件分析每张切片的阳性着色的积分吸光度值.结果：相对于癌旁组织,胃癌组织内RAGE和HMGB1的表达升高（P＜ 0.001）.RAGE的表达情况与肿瘤浸润深度、淋巴结转移及TNM分期显著相关（P值均＜0.05）; HMGB1的表达情况与肿瘤组织分化程度和肿瘤转移相关（P=0.023）; RAGE的表达与患者无病生存期相关（P＜0.001）,而HMGB1的表达与患者术后的无病生存期无关.结论：HMGB1-RAGE信号偶联与胃癌的侵袭和转移相关,患者胃癌标本中RAGE的表达影响患者预后,HMGB1-RAGE信号偶联是胃癌治疗的可能靶点之一.     

Advanced glycation end-product expression is upregulated in the gastrointestinal tract of type 2 diabetic rats

AIM:To investigate changes in advanced glycation end products(AGEs) and their receptor(RAGE) expression in the gastrointestinal(GI) tract in type 2 diabetic rats.METHODS:Eight inherited type 2 diabetic rats GotoKakizak(GK) and ten age-matched normal rats were used in the study.From 18 wk of age,the body weight and blood glucose were measured every week and 2 wk respectively.When the rats reached 32 wk,twocentimeter segments of esophagus,duodenum,jejunum,ileum,and colon were excised and the wet weight was measured.The segments were fixed in 10% formalin,embedded in paraffin and five micron sections were cut.The layer thickness was measured in Hematoxylin and Eosin-stained slides.AGE [N epsilon-(carboxymethyl) lysine and N epsilon-(carboxyethyl)lysine] and RAGE were detected by immunohistochemistry staining and image analysis was done using Sigmascan Pro 4.0 image analysis software.RESULTS:The blood glucose concentration(mmol/L) at 18 wk age was highest in the GK group(8.88 ± 1.87 vs 6.90 ± 0.43,P 0.001),a difference that continued to exist until the end of the experiment.The wet weight per unit length(mg/cm) increased in esophagus,jejunum and colon from the normal to the GK group(60.64 ± 9.96 vs 68.56 ± 11.69,P 0.05 for esophagus; 87.01 ± 9.35 vs 105.29 ± 15.45,P 0.01 for jejunum; 91.37 ± 7.25 vs 97.28 ± 10.90,P 0.05 for colon).Histologically,the layer thickness of the GItract was higher for esophagus,jejunum and colon in the GK group [full thickness(μm):575.37 ± 69.22 vs 753.20 ± 150.41,P 0.01 for esophagus; 813.51 ± 44.44 vs 884.81 ± 45.31,P 0.05 for jejunum; 467.12 ± 65.92 vs 572.26 ± 93.60,P 0.05 for colon].In esophagus,the AGE and RAGE mainly distributed in striated muscle cells and squamous epithelial cells.The AGE distribution was much stronger in the GK group compared to the normal group both in the striated muscle layer and mucosa layer(immuno-positive area/ total measuring area %:4.52 ± 0.89 vs 10.96 ± 1.34,P 0.01 for muscle; 8.90 ± 2.62 vs 22.45 ± 1.26,P 0.01 for mucosa).No visible difference was found for RAGE distribution between the two groups.In the intestine AGE and RAGE distributed in epithelial cells of villi and crypt.RAGE was also found in neurons in the myenteric and submucosal plexus.The intensity of AGE staining in mucosa of all segments and RAGE staining in neurons in all segments were strongest in the diabetes group.Significant difference for AGE was found in the epithelial cells of villi and crypt in duodenum(immunopositive area/total measuring area %:13.37 ± 3.51 vs 37.48 ± 8.43,P 0.05 for villi; 0.38 ± 0.12 vs 1.87 ± 0.53,P 0.05 for crypt) and for RAGE in neurons of all segments(e.g.,for jejunum:no staining neurons% 0 vs 0,mild 36.0 ± 5.2 vs 28.7 ± 3.5,moderate 53.2 ± 4.8 vs 55.8 ± 5.4,strong 10.7 ± 1.1 vs 15.4 ± 2.0,P 0.05).In the colon,RAGE was primarily found in neurons in the myenteric and submucosal plexus.It was stronger in the diabetes group than in the normal group(no staining neurons% 6.2 ± 0.2 vs 0.3 ± 0.04,mild 14.9 ± 2.1 vs 17.6 ± 1.5,moderate 53.1 ± 4.6 vs 44.7 ± 4.4,strong 25.6 ± 18 vs 43.6 ± 4.0,P 0.05).In the rectum,RAGE was primarily found in the mucosa epithelial cells.CONCLUSION:The AGE and RAGE expression was upregulated in the GI tract of GK diabetic rats and may contribute to GI dysfunction in type 2 diabetic patients.     

Advanced glycation end products induce mitochondrial pathway apoptosis in glomerular mesangial cells

Objective To investigate whether advanced glycation end products (AGEs) can induce the expression of Ros, JC-1 and its apoptosis-related proteins in glomerular mesangial cells under high glucose environment, induce apoptosis and injury of glomerular mesangial cells. Methods Rat glomerular mesangial cell line HBZY-1 was cultured in vitro. The cells were cultured with different concentrations of AGEs for 0, 12, 24 and 48 hours respectively. MTT assay was used to observe the cell proliferation ability. After the optimal time and concentration of AGEs were selected, the caspase enzyme inhibitor Z-VAD-fmk and reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine (NAC) were cultured and the apoptosis rate was detected by cell death detection apoptosis ELISA plus and Annexin V-FITC/PI kit. JC-1 staining was used to detect the changes of mitochondrial membrane potential (MMP). Cell ROX deep red flow cytometry was used to detect the total ROS level. The expression of anti-apoptotic protein Bcl-2, pro-apoptotic protein BAX, caspase-9, caspase-3 and poly ADP-ribose polymerase (PARP)-activated fragments was detected by Western blotting. Results AGEs could decrease the activity of glomerular mesangial cells in a time and concentration-dependent manner, and induce cell death. The percentage of apoptotic cells in glomerular mesangial cells was significantly increased after treatment with 250 mg/L AGEs for 24 h (P＜0.01), and Z-VAD-fmk could significantly alleviate AGEs-induced glomerular mesangial cell apoptosis (P＜0.01). Compared with the control group, AGEs increased the level of intracellular reactive oxygen species and decreased MMP in a time-dependent manner, and the two time points that AGEs significantly caused the change were 1 h and 2 h (all P＜0.01). AGEs also reduced the expression of antiapoptotic protein Bcl-2 and increased the expression of pro-apoptotic protein Bax, cleaved caspase-9, cleaved caspase-3 and cleaved PARP (all P＜0.01). Compared with AGEs group, NAC could significantly stabilize MMP (P＜0.01), increase Bcl-2 expression (P＜0.01), and decrease the expression of BAX, cleaved caspase-9, cleaved caspase-3 and cleaved PARP (all P＜0.01). Conclusion AGEs induce mitochondrial pathway apoptosis in glomerular mesangial cells by increasing intracellular ROS level and destroying MMP.     

Low molecular weight advanced glycation end products predict mortality in asymptomatic patients receiving chronic haemodialysis.

BACKGROUND: Advanced glycation end products (AGEs) have biological properties that may contribute to the premature cardiovascular mortality of haemodialysis patients. This study examines the hypothesis that low molecular weight forms of fluorescent AGEs (LMW fluorescence) predict mortality in haemodialysis patients. METHODS: The LMW fluorescence was measured in 85 patients treated with chronic haemodialysis and prospectively followed for 4 years. The primary outcome of all-cause mortality was assessed using Cox proportional hazards regression model. RESULTS: At the end of the follow-up period 37 (44%) patients died. The median LMW fluorescence level was 24.2 arbitrary units (range: 10.6-148.1 AU) and the receiver operator characteristic (ROC) curve cut-off for mortality was 37.0 AU. The LMW fluorescence predicted death both as a binary variable at the ROC cut-off, and as a continuous log-transformed variable when adjusted for age, albumin and C-reactive protein (CRP). Adjusted for age, albumin and CRP, the hazard ratio for mortality was 3.05 (1.41-6.60, P = 0.005) for LMW fluorescence as a binary variable and 2.71 per log unit (1.37-5.38, P = 0.004) as a continuous log-transformed variable. CONCLUSION: The low molecular weight forms of AGEs predict mortality in patients receiving chronic haemodialysis, and may be important in the mechanisms leading to atherosclerosis and inflammation in such patients.     

Impact of extracted urinary proteins and advanced glycation end product on autophagy pathway in renal tubular epithelial cells

Objective To investigate the effect of urinary proteins extracted from minimal change nephritic syndrome (MCNS) and advanced glycation end products (AGEs) on autophagy activity in renal tubular epithelial cells (TECs). Methods Kidney tissue specimens of patients with MCNS and DN were obtained from the kidney pathology library of the Affiliated Hospital of Guangdong Medical College. The kidney tissue from patients with hematuria and proven to be minimal change by pathology examination were used as control. The expression of LC3-Ⅱ in kidney was examined by immune histochemistry in vivo. Expression of LC3-Ⅱ was also studied after exposing HK-2 cells to 8 g/L urinary proteins and 100 mg/L AGE-BSA respectively. LC3-Ⅱ turnover was examined after exposure to urinary proteins in presence of Lysosomal inhibitors leupeptin (200 mg/L) or chloroquine(10μmol/L) by western blot assay. In addition, the autophagosome or autolysosome formation was assessed after transfecting a tandem mRFP-GFP tagged LC3 (tfLC3) plasmid into HK-2 cells. Finally, the production of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) were measured by ELISA after exposure of cells to autophagy enhancer rapamycin (10 μmol/L) and autophagy inhibitor chloroquine (10 μmol/L) in addition to urinary proteins. Results (1)In comparison with the control group, the expression of LC3-Ⅱ was significantly increased in TECs from patients with MCNS and DN(P＜0.01). (2)The expression of LC3-Ⅱwas enlarged after exposed to urinary proteins(P＜0.01), and further increased after leupeptin (autophagy inhibitor) addition. (3) Exposure to urinary proteins increased the autophogosomes and autolysosomes when observed by transfection of tfLC3 plasmid(P＜0.01). (4)The expression of LC3-Ⅱ was also elevated after treatment with AGE-BSA(P＜0.01), but no further increase after chloroquine (autophagy inhibitor) addition. (5) Only the autophogosome formation(P＜0.01), but not autolysosome formation, was found increased by transfection of tfLC3 plasmid after exposure to AGE-BSA. (6) Pre-treatment of HK-2 cells with autophagy enhancer rapamycin reduced the productions of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1), while blocking autophagy with autophagy inhibitor chloroquine exerted an opposite effect. Conclusions Autophagy was activated by urinary proteins, but inactivated by AGE-BSA. Autophagy activation may play a key role in protecting TECs in the progression of primary and secondary kidney diseases.     

Immunohistochemical detection of advanced glycation end products in human bladder with specific monoclonal antibody

Objective:   To investigate the accumulation of advanced glycation end products (AGE) in human bladder.
Methods:   Human bladder specimens were obtained from nine patients during radical cystectomy. Frozen sections were immunohistochemically analyzed by three different monoclonal anti-AGE antibodies such as anti-N^ε-(carboxymethyl)lysine (CML), anti-imidazolone and anti-pentosidine antibodies. Bladder sections were stained with these antibodies by indirect immunoperoxidase methods. Double immunohistochemical staining with one of the anti-AGE antibodies or an anti-human macrophage antibody was also carried out.
Results:   We demonstrated that CML and pentosidine were accumulated in human bladder extracellularly as well as intracellularly, whereas any accumulation of imidazolone was not observed. Double immunohistochemical staining indicated that AGE-accumulated cells in human bladder were derived from macrophages.
Conclusions:   The present study demonstrated that AGE-structures such as CML and pentosidine are accumulated extracellularly in human bladder, and were endocytosed by tissue macrophages.     

糖基化终末产物对人外周血内皮祖细胞生物学特性的影响

目的 观察糖基化终末产物(AGEs)对体外培养的人外周血内皮祖细胞(EPCs)生物学特性的影响.方法 以密度梯度离心法获取人外周血单个核细胞(MNCs),由激光共聚焦显微镜鉴定FITC-UEA-1和Dil-acLDL双染色阳性细胞为正在分化的EPCs.加入不同浓度AGEs培养48h,然后分别采用MTT法、Boyden小室测定来观察EPCs的增殖、迁移能力;以人纤维连接蛋白(hFN)检测EPCs的黏附能力;分别采用甲醛和Dnase Ⅰ诱导EPCs凋亡作为阳性对照组,用AnnexinV-FITC/PI和TUNEL法流式细胞仪检测AGEs对EPCs凋亡率的影响.结果 高浓度AGEs可减少EPCs贴壁细胞数量(P<0.01),明显减弱EPCs的黏附(P<0.01)、增殖(P<0.001)、迁移能力(P<0.05),提高EPCs的早期凋亡率(P<0.001).结论 AGEs可减少EPCs的数昔并使EPCs的功能受损.