胰岛素对高糖环境下大鼠颌骨骨髓基质细胞成骨分化过程中peroxiredoxin-6表达的影响

Effect of insulin on peroxiredoxin-6 in the osteogenic differentiation of rat's mandiular bone marrow stromal cells in high glucose

目的:探讨胰岛素对高糖环境下大鼠颌骨骨髓基质细胞(rat bone marrow stromal cells,rBMSCs)成骨分化过程中peroxiredoxin-6表达的影响。方法:从Wistar大鼠下颌骨中分离培养骨髓基质细胞,分别于不同糖浓度(5.5、25、45 mmol/L)及胰岛素(0、10-5mol/L)的干预下成骨诱导(5.5 mmol/L组、5.5 mmol/L+Ins组、25 mmol/L组、25 mmol/L+Ins组、45 mmol/L组、45 mmol/L+Ins组)。成骨诱导21 d后,进行茜素红染色,矿化结节定量分析。成骨诱导1、3、7、14、21 d后,用ELISA试剂盒检测peroxiredoxin-6蛋白的表达。采用SPSS15.0软件包对数据进行统计学分析。结果:45 mmol/L高糖环境显著抑制rBMSCss的矿化,胰岛素可改善高糖对rBMSCs矿化的抑制。45 mmol/L组peroxiredoxin-6的表达较5.5 mmol/L组和25 mmol/L组均显著下降。矿化诱导第21天,各组peroxiredoxin-6蛋白的表达均达到最高峰。在25 mmol/L和45 mmol/L的糖浓度下,10-5mol/L的胰岛素可显著上调rBMSCs peroxiredoxin-6蛋白的表达。结论:高糖环境抑制peroxiredoxin-6蛋白的表达,胰岛素可改善高糖环境对peroxiredoxin-6蛋白在大鼠颌骨骨髓基质细胞成骨分化过程中表达的影响。Peroxiredoxin-6蛋白在大鼠颌骨骨髓基质细胞成骨分化后期表达显著。

PURPOSE： To explore the effect of insulin on the expression of peroxiredoxin-6 in osteogenic differentiation of rat＇s mandibular bone marrow stromal cells （rBMSCs） in high glucose. METHODS： Bone marrow stromal cells were obtained from the mandible of Wistar rats and stimulated in three glucose concentrations mineral medium （5.5 mmol/L, 25 mmol/L and 45 retool/L） with or without insulin （10-5mol/L） for 1, 3, 7, 14, and 21 days. The expression of prohibitin was quantified via enzyme-linked immuno sorbent assays （ELISA）. The mineralization nodules were assessed at day 21 by alizarine red staining. Statistical analysis was performed using SPSS 15.0 soft ware package. RESULTS： High glucose of 45 mmol/L inhibited mineralization of rBMSCs and insulin can improve the mineralization in high glucose. The expression of peroxiredoxin-6 in 45 mmol/L group decreased significantly compared with 5.5 mmol/L group and 25 mmol/L group. The expression of peroxiredoxin-6 in each group achieved maximum at day 21. Insulin （10-5 mol/L） increased the expression of peroxiredoxin-6 in 25 mmol/L group and 45 mmol/L group in osteogenic differentiation of rBMSCs. CONCLUSIONS： High glucose inhibits the expression of peroxiredoxin-6 in osteogenic differentiation of rBMSCs, while insulin upregulates the expression of peroxiredoxin-6 in rBMSCs. Peroxiredoxin-6 may play an important part in later stage in osteogenie differentiation of rBMSCs.