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糖化低密度脂蛋白对成骨细胞低氧诱导因子1α表达的影响

ISSN号：1006-6187
期刊名称：《中国糖尿病杂志》
时间：0
分类：R587.2[医药卫生—内分泌;医药卫生—临床医学;医药卫生—内科学]
作者机构：[1]同济大学附属同济医院内分泌科,上海200065, [2]上海中医药大学附属普陀医院内分泌科
相关基金：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）

中文摘要：

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。

英文摘要：

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.

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