高糖对人骨髓间充质干细胞成骨能力影响的体外研究

Osteogenic Responses of Human Bone Mesenchymal Stem Cells to Hyperglycemia in vitro

目的:体外模拟糖尿病病人高血糖状态,观察高糖对人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)成骨能力的影响。方法 :将使用不同含糖量培养液体外培养的hMSCs分为5.5mmol/L生理糖浓度组、25mmol/L高糖组、44 mmol/L高糖组。CCK-8试剂盒检测各组hMSCs的增殖速度;骨诱导7、14、21 d时分别观察高糖对hMSCs骨向分化相关基因骨钙素(OCN)、骨桥蛋白(OPN)、Runx相关因子2(Runx-2)的表达,碱性磷酸酶(ALP)活性,钙沉积量的影响。结果:两个高糖组相对生理糖组,有抑制hMSCs的增殖能力(P<0.05),且抑制ALP活性及细胞外钙基质沉积,下调了骨向分化相关基因OCN、OPN、Runx-2表达水平(P<0.01),该抑制作用随糖的浓度升高而加强。结论:高糖显著抑制了hMSCs生物矿化过程,并以浓度依赖的方式降低hMSCs的增殖及成骨能力。

Objective： The current study designed to examine whether human bone mesenchymal stem cells （hMSCs） modulate early osteogenic metabolism during exposure to hyperglycemia condition in vitro. Methods： Third-passage hMSCs were used for experiments and divided into 3 groups： 5.5 mmol/L normal glucose control group,25 mmol/L high glucose group, and 44 mmol/L high glucose group. Cell proliferation was measured by counting kit （CCK）-8. The effects of high glucose on hMSCs osteogenesis related gene OCN,OPN and Runx2 mRNA expression,ALP activity,calcium deposits were measured at days 7, 14, and 21. Results： In higher glucose concentration （25 and 44 mmol/L） groups, hMSCs proliferation, ALP activity, calcium deposition, OCN,OPN,Runx2 mRNA expression were significantly reduced as compared with 5.5 rnmol/L normal glucose group. High glucose showed an inhibition effect on activities of hMSCs in a concentration-dependent manner. Conclusion： High glucose inhibits biomineralization process in hMSCs; High glucose may inhibit the proliferation and differentiation of hMSCs in a concentration-dependent manner.