高糖对小鼠脑微血管内皮细胞释放高迁移率蛋白-1的影响

The influence of hyperglycemia on releasing high-mobility group protein-1 of cerebral microvascular endothelial cells

目的研究不同浓度高糖培养基对小鼠脑微血管内皮细胞(bEnd.3)的损伤作用。方法对照组:25 mmol/L糖浓度组;高糖诱导组:35、40、45、50、60 mmol/L糖浓度组;辛伐他汀干预组:高糖(40 mmol/L)+辛伐他汀(0.1μmol/L)、高糖(40 mmol/L)+辛伐他汀(1μmol/L)、高糖(40 mmol/L)+辛伐他汀(10μmol/L)。将各实验组分别培养10、18、24、36、48、72 h用于试验,辛伐他汀干预组培养24 h后用于实验。细胞酶联免疫吸附法(ELISA)测定各组细胞上清液中高迁移率蛋白-1(HMGB-1)值,光镜和电镜观察细胞形态和结构的改变。结果在同一时间点测得HMGB-1进行比较,可发现各实验组随葡萄糖浓度的升高HMGB-1释放量呈递增趋势,培养48 h后,高糖组(40、45 mmol/L)达到释放峰值;辛伐他汀干预组释放HMGB-1量明显下降。各高糖组可引起细胞线粒体明显肿胀,空泡样变,以60 mmol/L糖浓度组最为明显;辛伐他汀干预组细胞线粒体结构则无明显变化。结论高糖可引起内皮细胞损伤,其潜在的机制很有可能是通过晚期炎症因子HMGB-1的释放而起到炎症介导的的过程。

Objective To study the impairment effect of the difference high glucose concentration to mouse cerebral microvascular endothelial cells（bEnd.3）in vitro.Methods The cells were divided into control group（25mmol/L glucose）;high glucose-induced group（five sub-group：35,40,45.50,60mmol/L glucose）and simvastatin interventional group（three concentration sub-groups：40mmol/L high glucose＋simvastatin 0.l,1.0,l0μmol/L）.The first two group were incubated for 10,18,24,36,48,72 hour and the third group for 24 hour.Then the High-mobility group protein-1 release by cerebral microvascular endothelial cells in supernatant medium of every was detected group by Enzyme-linked immunosorbent assay（ELISA）.Results Compare the level of the High-mobility group protein-1 in the same time,the secretion of HMGB-1 was liberated more as the glucose concentration was steped up.After Incubated for 48 hour,compared with the control group,the group of the 40,45mmol/L high glucose released the predominance content of HMGB-1;The HMGB-1 releasing of MVECs in each high glucose﹢simvastatin interventional group declined obviously.High glucose concentrations caused cell body swelling,mitochondrial swelling and vacuolar degeneration,especially in 60mmol/L glucose group,compared with the control group;The changes of cell body swelling,mitochondrial swelling and vacuolar degeneration,in 40mmol/L high glucose﹢simvastatin group were not so obvious.Conclusion High glucose can cause endothelial cell＇s injury,besides,the secretion of HMGB-1 is gradually increase as the time,its potential mechanisms may be related to the releasing of advanced stage inflammatory mediators HMGB-1 to mediate the process of inflammation.