高糖对人骨髓间充质干细胞成骨分化的影响

目的：探讨不同葡萄糖浓度培养环境对人骨髓间充质干细胞（human bone marrow mesenchymal stem cells，hBMSC）增殖和成骨分化的影响。方法用含不同葡萄糖浓度的基础培养基培养，在1、4、7、10 d进行CCK?8细胞增殖检测；用含不同葡萄糖浓度的成骨诱导分化培养基培养细胞7 d，观察hBMSC分化情况；实时荧光定量PCR法检测碱性磷酸酶（alkaline phosphatase，ALP）、骨钙素（osteocalcin，OC）、I型胶原蛋白（collagen type I， Col?1）基因表达水平；在7 d进行钙茜素红染色显示矿化结节形成。结果10 mM葡萄糖有助于hBMSC细胞增殖，高浓度葡萄糖（＞30 mM）能够抑制hBMSC增殖（P＜0.05）；成骨诱导液能诱导hBMSC成骨分化，但葡萄糖浓度升高会减少hBMSC的矿化结节形成、抑制成骨标志基因ALP、OC和Col?1表达（P＜0.05）。结论在高糖环境培养下，抑制hBMSC增殖、下调成骨标志性基因Col?1、ALP、OC的表达，同时高糖可以减弱干细胞的成骨矿化能力，间接影响骨形成和代谢。

Effects of high glucose on osteogenic differentiation of hBMSC

Objective To investigate the effects of different glucose concentration on the proliferation and osteogen?ic differentiation of human bone marrow mesenchymal stem cells (hBMSC) in vivo. Methods Cultured with basal medi?um containing different glucose concentrations, CCK?8 cell proliferation was detected at 1, 4, 7, 10 days. The osteogenic differentiation of human bone marrow mesenchymal stem cells was observed at 7 d, which was induced by osteogenic differentiation medium with different concentration of glucose. The expressions of alkaline phosphatase (ALP), osteocal?cin (OC) and collagen type I (Col?1) gene were detected by real?time fluorescence quantitative PCR. Mineralized nodule formation was displayed by calciumalizarin red staining on the seventh day. Results 10 mM glucose stimulated prolif?eration of hBMSC, while the higher (>30 mM) inhibited the proliferation (P < 0.05); Osteogenic induction can induce osteogenic differentiation of hBMSC, but the increase of glucose concentration will decrease the formation of mineralized nodules of hBMSC, inhibit the expression of osteogenic marker genes ALP, OC and Col?1 (P<0.05). Conclusion The expression of Col?1, ALP and OC in osteoblast was down?regulated by high glucose, and the hBMSC proliferation was in?hibited. At the same time, high glucose can reduce the osteogenic mineralization ability of stem cells and indirectly af?fect bone formation and metabolism.