文档介绍：华中科技大学
硕士学位论文
高糖对血管内皮细胞表达HMGB1及对氧化应激的作用研究
姓名:赵杨
申请学位级别:硕士
专业:神经病学
指导教师:刘昌勤
2011-05
华中科技大学硕士学位论文

高糖对血管内皮细胞表达HMGB1 及对氧化应激的作用研究

华中科技大学同济医学院附属协和医院神经内科
硕士研究生:赵杨
导师:刘昌勤

中文摘要

目的探讨高糖对血管内皮细胞分泌高迁移率族蛋白B1(HMGB1)的影响,以及高糖条
件培养液对血管内皮细胞氧化应激水平的影响。
方法 (正常对照组),20mmol/L( 高糖组 1),
33mmol/L(高糖组 2)的DMEM培养基,体外培养人脐静脉血管内皮细胞株(),
台盘兰排斥法检测 48h以内细胞的存活率,使用Western blot检测培养液上清中HMGB1
的表达水平。 33mmol/LDMEM高糖培养基, 混合液(高糖组 2 的上清液与
33mmol/LDMEM高糖培养基混合),刺激血管内皮细胞,采用分光光度比色法检测血管内
皮细胞内丙二醛(MDA)和超氧化物岐化酶(SOD)的水平。
结果 ,高糖组培养液上清中HMGB1 表达均明显增加(P<)。
台盼蓝排斥法检测 48h以内各组细胞生存率均>95%,各组间生存率无明显差异。2.
与单纯高糖培养基刺激组相比,混合液刺激组的血管内皮细胞内MDA含量明显升高,
而SOD活力则显著下降(p<)。
结论高糖可诱导体外血管内皮细胞表达HMGB1。高糖环境下,机体可能通过增加分泌
HMGB1,加强氧化应激反应而促进血管内皮细胞损伤。

【关键词】高糖; 血管内皮细胞;高迁移率族蛋白 B1 ;氧化应激

2
华中科技大学硕士学位论文

A study on expression of HMGB1 in vascular endothelial cells
and the effect on oxidative stress by high glucose

Department of neurology, Union Hospital, Tongji Medical College
Huazhong University of Science Technology
Master Candidate: Zhao Yang
Supervisor: Prof. Liu Changqin

Abstract
Objective: To observe the effects of high glucose on the expression of HMGB1 in cultured
vascular endothelial cells (VECs) and the effect of the culture fluid with high glucose
condition on oxidative stress.
Methods 1. The human umbilical veins endothelial line () were cultured in vitro.
VECs were cultured in medium with different glucose concentrations for 48h. The glucose
concentrations were mmol/L (normal control), 20 mmol/L (high glucose group 1), and
33mmol/L (high glucose group 2) .The levels of HMGB1 in the supernatant were measured
by Western blot. The cell viability within 48h was examined by trypan blue exclusion test.
2. VECs were divided into two groups: high glucose group (33mmol/L) and the mixed
group (the supernatant of high glucose group 2 plus 33mmol/L high glucose medium). 24
hours later, th