Li Yu-ming, Deng Bo, Li Li-hua, Deng Li-ping, Gao Hai-bo, Chen Lu-lu
Department of Endocrinology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Zhonghua Yi Xue Za Zhi. 2009 Sep 29;89(36):2583-5.
To observe the effects of different concentrations of glucose and insulin in cell differentiation from bone marrow stem cells (BMSC) into osteoblasts (OB) and explore the acting mechanism of glucose and insulin in bone metabolism.
The in vitro technique of cell culture was employed to separate, purify and cultivate BMSC from the tibia and femur of rats. Then the BMSCs were differentiated into osteoblasts under the inductions of different glucose concentrations (5.6, 25, 50 mmol/L) with or without the addition of insulin (0.6 microg/ml). Later the calcium nodule cell ratio of osteoblasts in alizarin red stain was observed microscopically to assay the mRNA expression of OB markers-ALP, BGP and Runx2 by real-time PCR.
With the increasing glucose concentrations, the number of red calcium nodules obviously decreased. PCR showed that the mRNA expression of ALP, BGP and transcription factor Runx2 decreased too. The difference was significant (P < 0.05). The differentiation of OB decreased with with the increasing glucose concentrations. The cell number of red calcium nodules obviously increased in the same glucose concentration groups. The mRNA expression of ALP, BGP and Runx2 also increased (P < 0.01). It hinted that insulin could enhance the differentiation of OB.
Glucose can inhibit the differentiation from BMSC into OB. The inhibition is more sensitive along with the concentration increments. This may be one of the pathogenesises of diabetic osteoporosis. Insulin can enhance the differentiation from BMSC into OB. In addition, it can improve the inhibition of differentiation from BMSC into OB because of a high concentration of glucose.
观察不同浓度葡萄糖和胰岛素对大鼠骨髓干细胞(BMSC)向成骨细胞(OB)分化的影响,探讨葡萄糖和胰岛素在骨代谢中的作用机制。
采用细胞培养体外技术分离、纯化和培养大鼠胫骨和股骨的BMSC。然后在不同葡萄糖浓度(5.6、25、50 mmol/L)诱导下,添加或不添加胰岛素(0.6 μg/ml),将BMSC诱导分化为成骨细胞。之后通过茜素红染色显微镜观察成骨细胞钙结节细胞比例,采用实时荧光定量PCR检测OB标志物碱性磷酸酶(ALP)、骨钙素(BGP)和Runx2的mRNA表达。
随着葡萄糖浓度升高,红色钙结节数量明显减少。PCR结果显示,ALP、BGP和转录因子Runx2的mRNA表达也降低。差异有统计学意义(P < 0.05)。随着葡萄糖浓度升高,OB分化减少。在相同葡萄糖浓度组中,红色钙结节细胞数量明显增加。ALP、BGP和Runx2的mRNA表达也增加(P < 0.01)。提示胰岛素可促进OB分化。
葡萄糖可抑制BMSC向OB分化,且随着浓度升高抑制作用更敏感。这可能是糖尿病性骨质疏松症的发病机制之一。胰岛素可促进BMSC向OB分化,此外,还可改善高浓度葡萄糖对BMSC向OB分化的抑制作用。