基础医学与临床 ›› 2012, Vol. 32 ›› Issue (10): 1212-1215.

• 研究论文 • 上一篇    下一篇

维持性血液透析患者血清诱导人脐静脉内皮细胞凋亡

王喜超,涂阳科,吕凤岩   

  1. 天津市第一中心医院肾内科
  • 收稿日期:2011-10-08 修回日期:2012-03-10 出版日期:2012-10-05 发布日期:2012-09-28
  • 通讯作者: 王喜超 E-mail:wxichao@sohu.com

Hemodialysis patients serum induce apoptosis of HUVECs

  • Received:2011-10-08 Revised:2012-03-10 Online:2012-10-05 Published:2012-09-28

摘要: 目的 研究血液透析患者血清诱导人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)的凋亡,并初步探讨其机制。方法 取透析患者血清(实验组)及健康志愿者血清(对照组)培养HUVECs 30和40 h,倒置显微镜下观察形成血管样结构的内皮细胞的凋亡,MTT法检测内皮细胞增殖,TUNEL法检测内皮细胞凋亡,比色法检测内皮细胞内活性氧产生,Westernblot法检测内皮细胞内caspase-3表达。结果 内皮细胞培养40h后,实验组形成血管样结构的内皮细胞出现凋亡(P<0.05),内皮细胞ROS产生增加(P<0.05),以及内皮细胞caspase-3表达量为(0.83±0.03),显著高于对照组(0.57±0.02)(P<0.05)。结论 血液透析患者血清可以诱导形成血管样结构的内皮细胞凋亡,其机制可能与内皮细胞内caspase-3表达增加以及ROS产生增多有关。

关键词: 血液透析, 内皮细胞, 凋亡, 血管新生

Abstract: Objective To validate the apoptosis of HUVECs induced by hemodialysis patients seum and explore the mechanism.Methods HUVECs are exposed to culture medium containing serum obtained from patients undergoing haemodialysis(trial group) and healthy volunteers(control group).The effect of uraemic serum on the apoptosis of endothelial cells in vascular networks are observed under light-microscope;Cell proliferation are measured by MTT;Apoptotic cells in vascular networks are evaluated by TUNEL;To investigate the expression of caspase-3 and production of ROS, cell lysates are evaluated by western blotting and spectrophotometry.Results After HUVECs are exposed to pooled serum medium for 40 h, there are apoptotic endothelial cells in vascular networks (P<0.05) and the endothelial cells produce more ROS in trial group(P<0.05).At 40h, the expression level of caspase-3 in endothelial cells in trial group is(0.83±0.03),significantly higher than in control group(0.57±0.02)(P<0.05).Conclusion Hemodialysis patients serum can induce apoptosis of HUVECs in vascular networks. Higher expression of caspase-3 and more production of ROS in endothelial cells may be involved in the mechanism.

Key words: hemodialysis, endothelial cells, apoptosis, angiogenesis

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