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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 study the effects of low density lipoprotein (LDL) with different saccharification on the proliferation and metabolism of osteoblasts and to explore its possible mechanism. Methods Mouse osteoblasts (MC3T3-E1) were treated with different concentrations of glycosylated LDL (4.97% and 8.24%) for 48 h. Normal LDL (n-LDL) group and blank control group The expression of HIF-1α, VEGF mRNA and HIF-1α protein were determined. The cell viability and osteocalcin (OC) level were measured. Results The cell viability of n-LDL group, 4.97% gly-LDL group and 8.24% gly-LDL group were 95.6%, 57.1% and 71.2% respectively. Compared with the Blank group, the experimental group OC decreased, and with the LDL glycosylation, OC greater inhibition of secretion. Compared with Blank group, the expression of VEGF mRNA in n-LDL group was down-regulated (P <0.05). Compared with n-LDL group, VEGF expression increased in 8.24% gly-LDL group. Compared with blank group and n-LDL group, 8.24% gly-LDL up-regulated the expression of HIF-1α mRNA (P <0.01) and up-regulated the protein expression of HIF-1α. Conclusion LDL with high saccharification can affect the gene and protein expression of HIF-1α in mouse osteoblasts, which may be one of the mechanisms by which gly-LDL affects the proliferation and metabolism of osteoblasts.