泛素在高糖诱导的血管内皮细胞凋亡中的表达  

Expression of Ubiquitin mRNA in Apoptotic Endothelial Cells Induced by High Concentration of Glucose

作　　者：王晓[1] 李果[1] 李纪平[1] 刘赟[1] 

机构地区：[1]上海第二医科大学瑞金医院上海市内分泌代谢病研究所,上海200025

出　　处：《上海第二医科大学学报》2005年第1期36-38,共3页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金(30100085)国家自然科学基金面上项目(30370667)资助.

摘　　要：目的检测高浓度葡萄糖对血管内皮细胞凋亡的影响和泛素表达的变化。方法ECV-304细胞在高糖(25mmol/L)和正常糖浓度(5.5mmol/L)DMEM培养液中分别培养24、48、72h,用MTT法检测细胞活力,用琼脂糖凝胶电泳检测细胞凋亡特异性的DNA片段,并用Northernblot检测泛素mRNA的变化。结果高糖24、48、72h组细胞OD值比正常浓度组均显著下降(P<0.05);1.8%琼脂糖凝胶电泳在高糖48、72h组检测到细胞凋亡特异性DNA片段;泛素mRNA在高糖24、48、72h组分别增加25.37%、53.47%和63.93%(P<0.05)。结论高浓度葡萄糖可增加血管内皮细胞凋亡,泛素mRNA在高糖诱导的内皮细胞凋亡中表达上调,且发生在凋亡形态学改变之前,泛素参与调节高血糖诱导的细胞凋亡。Objective To establish a model of high-glucose-triggered endothelial cell apoptosis, then determine the expression of ubiquitin mRNA. Methods Human umbilical vein endothelial cell line ECV-304 was exposed to 25 mmol/L and 5. 5 mmol/L glucose for different time periods. Cell viability was measured by MTT test, gel electro-phoresis was carried out for assaying DNA fragmentation, the expression of ubiquitin mRNA were measured by Northern blotting. Results The OD value of MTT test in ECV-304 cells cultured in 25 mmol/L glucose for 24, 48, 72 h decreased in comparison with those of 5. 5 mmol/L glucose ( P < 0. 05 ) ; the typical ladder pattern of DNA fragmentation was found in those cells treated with 25 mmol/L glucose for 48 h or 72 h in DNA gel electrophoresis; the level of ubiquitin mRNA in ECV-304 cells cultured in 25 mmol/L glucose for 24, 48, 72 h increased by 25.37% , 53.47% and 63.93% respectively compared with the control group (P <0.05). Conclusion High glucose may induce ECV-304 cells apoptosis. Ubiquitin mRNA expression increased even before the detection of apoptosis. Ubiquitin may have an important role in high-glucose-triggered endothelial cell apoptosis.

关 键 词：高糖 内皮细胞 泛素 细胞凋亡 表达 糖尿病 血管内皮细胞功能紊乱 

分 类 号：R587.2[医药卫生—内分泌]