摘要
目的研究不同糖化程度的低密度脂蛋白(LDL)对成骨细胞增殖及代谢的影响并探讨其可能机制。方法用不同程度(4.97%和8.24%)的糖化LDL(gly-LDL)处理小鼠成骨细胞(MC3T3-E1)48 h,设普通LDL(n-LDL)组和空白对照(Blank)组,测定缺氧诱导因子-1α(HIF-1α),VEGF mRNA表达,以及HIF-1α蛋白表达,同时检测细胞存活率及细胞上清骨钙素(OC)水平。结果 n-LDL组、4.97%gly-LDL组和8.24%gly-LDL组细胞存活率分别为95.6%、57.1%和71.2%。与Blank组比较,各实验组OC降低,且随着LDL的糖化,OC分泌受到抑制的程度越大。n-LDL组与Blank组比较,VEGFmRNA表达量下调(P<0.05);8.24%gly-LDL组与n-LDL组比较,VEGF表达量相对上调。与Blank组及n-LDL组比较,8.24%gly-LDL上调HIF-1αmRNA的表达(P<0.01),同时上调HIF-1α的蛋白表达。结论高糖化程度的LDL可影响小鼠成骨细胞HIF-1α的基因及蛋白表达影响,这可能是gly-LDL影响成骨细胞增殖与代谢的机制之一。
Objective To explore the impact of different levels of glycated low density lipoprotein（gly-LDL） on osteoblast proliferation and metabolism and to discuss the possible mechanism.Methods Mouse osteoblasts（MC3T3-E1） were treated with different degrees of saccharifying gly-LDL（4.97%and8.24%） for 48 hours.Normal LDL（n-LDL） group and blank group were set as control.Real time PCR was used to test the mRNA expression of HIF-1 alpha and VEGF.SDS-PAGE method was taken to test the protein expression of HIF-1 alpha.The influence of gly-LDL on the proliferation and metabolism of osteoblast was measured by the way of CCK8 and ELISA.Results The cell survival rates of n-LDL group,4.97%and 8.24%gly-LDL group were 95.6%,57.1%and 71.2%,respectively.The OC levels of experimental groups were significantly lower than control group,with the trend of the higher glycated level,the less OC secreted.Compared with control group,the mRNA expression of VEGF in n-LDL group was down-regulated（P〈0.05）.Compared with n-LDL group,the mRNA expression of VEGF in 8.24%gly-LDL group was significantly upregulated,but there was no significant difference between 8.24%glyLDL group and contol group.Compared with n-LDL group and blank group,the mRNA expression of HIF-1α in 8.24%gly-LDL osteoblast was significantly upregulated,and the protein expression of HIF-1αin 8.24%gly-LDL osteoblast was downregulated.Conclusion High level glycated LDL may influence the expression of HIF-1α in MC3T3-E1,which may be one of the mechanisms for osteoblast proliferation and metabolism.
作者
张冰雨
曾瑞翔
朱淇
单畅
雷涛
ZHANG Bing-yu ZENG Rui-xiang ZHU Qi et al(Department of Endocrinology, Tongji Hospital of Tongji University, Shanghai 200065, Chin)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2017年第2期154-158,共5页
Chinese Journal of Diabetes
基金
863科研专项基金(2014AA022304)
上海市卫生局科研课题(20124256)
关键词
低密度脂蛋白
成骨细胞
低氧诱导因子1Α
血管内皮生长因子
Low density lipoprotein（LDL）
Osteoblast
Hypoxia-inducible factor 1-α（HIF-1α）
Vascular endothelial growth factor（VFGF）
