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。 Objective: To investigate the effects of advanced glycation end products (AGEs) on the proliferation and apoptosis of human vascular endothelial cells and explore the mechanism of neovascularization or delay in the treatment of diabetic refractory wounds. METHODS: AGE-modified human serum albumin (AGE-HSA) and human vascular endothelial cells (ECV304) were co-cultured in vitro with different time and concentration of AGE-HSA. MTT colorimetric assay and cell counting were used to detect AGE-HSA Inhibition of proliferation of vascular endothelial cells, and trypan blue exclusion test to detect cell viability. FITC Annexin-V and propidium iodide (PI) staining were used to detect the percentage of apoptotic cells by flow cytometry. Fluorescence confocal microscope was used to observe the fluorescence staining of FITC Annexin-V and PI in apoptotic or dead cells. The morphological changes of apoptotic cells were observed by electron microscope and light microscope. Results: Under the conditions of 12.5, 25 and 50μg / ml AGE-HSA, the proliferation of endothelial cells on the 2nd day was not significantly different from that of the control group, but significantly lower than that of the control group on the 4th and 6th day (P < 0.01). The endothelial cells were exposed to 100 or 200μg / ml AGE-HSA for 6h. The OD value of endothelial cells was significantly different from that of the control group (P <0.01) by MTT assay. At the same time, the endothelial cells showed specific apoptotic morphological changes and The number of FITC Annexin-V-positive and / or PI-positive cells gradually increased, and the number of apoptotic or dead cells increased with the time and concentration of AGE-HSA. CONCLUSION: AGE-HSA can inhibit the proliferation of endothelial cells and induce apoptosis of AGE-HSA, which is related to the time and concentration of AGE-HSA. It is suggested that AGEs may be one of the mechanisms that cause the neovascularization disorder or delay in the wound healing of diabetic patients.