高糖对人骨髓间充质干细胞成骨分化的影响

Effects of high glucose on osteogenic differentiation of hBMSC

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 stemcells,h BMSC)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h BMSC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 m M葡萄糖有助于h BMSC细胞增殖,高浓度葡萄糖(>30 m M)能够抑制h BMSC增殖(P<0.05);成骨诱导液能诱导h BMSC成骨分化,但葡萄糖浓度升高会减少h BMSC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h BMSC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。

Objective To investigate the effects of different glucose concentration on the proliferation and osteogen-ic differentiation of human bone marrow mesenchymal stem cells（h BMSC） 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 osteogenicdifferentiation of human bone marrow mesenchymal stem cells was observed at 7 d, which was induced by osteogenicdifferentiation 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 noduleformation was displayed by calciumalizarin red staining on the seventh day. Results 10 m M glucose stimulated prolif-eration of h BMSC, while the higher（〉30 m M） inhibited the proliferation（P〈0.05）; Osteogenic induction can induceosteogenic differentiation of h BMSC, but the increase of glucose concentration will decrease the formation of mineralizednodules of h BMSC, inhibit the expression of osteogenic marker genes ALP, OC and Col-1（P〈0.05）. Conclusion Theexpression of Col-1, ALP and OC in osteoblast was down-regulated by high glucose, and the h BMSC 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.