葡萄糖对大鼠骨髓基质干细胞向成骨细胞分化的影响
Effects of glucose on bone marrow stromal cells differentiation into osteoblastic lineages
目的 体外观察不同葡萄糖浓度在促骨髓基质干细胞向成骨细胞分化过程中对细胞因子碱性磷酸酶、骨形成蛋白-2、转化生长因子β1表达的影响,并探讨其对骨代谢的作用机制.方法 无菌条件下从2周龄SD大鼠长骨骨髓中分离获取骨髓基质干细胞,采用全骨髓贴壁培养法对骨髓基质干细胞进行纯化、传代扩增,随后在不同糖浓度(5.5 mmol/L、25.0 mmol/L)干预下向成骨细胞诱导分化培养21 d,进行茜素红染色观察矿化结节并测定成骨细胞的标记物碱性磷酸酶、骨形成蛋白-2及转化生长因子β1表达,比较各组中成骨细胞分化的情况.组间比较采用单因素方差分析法.结果 25.0 mmol/L糖浓度组与5.5 mmol/L糖浓度组比较,钙结节形成比例分别为(45.3±0.7)%、(68.3±0.8)%,差异具有统计学意义(P<0.05),碱性磷酸酶(0.350±0.020、0.563±0.043)、骨形成蛋白-2[(590±27)、(744±41)μg/L]及转化生长因子β1[(875±40)、(1188±52)μg/L]活性比较,差异具有统计学意义(均P<0.05).结论 在高糖条件下,骨髓基质干细胞向成骨细胞的分化减弱,这可能是糖尿病性骨质疏松的重要机制之一.
更多Objective To observe the effects of glucose on the expression of alkaline phosphatase ( ALP), bone morphogenetic protein-2 ( BMP-2 ) and transforming growth factor-beta 1 ( TGF-51 ) for rat bone marrow stromal cells (BMSCs) differentiation into osteoblastic(OB) and to investigate the mechanism of the impact on the bone metabolism from them. Methods Bone-marrow stromal cells were achieved from long bone marrow of rats under aseptic condition. After purification, passage and amplification by using the whole bone marrow adherence method, the BMSCs were interfered into osteoblasts with different doses of glucose concentration (5.5 mmol/L and 25.0 mmol/L) in the presence of the osteogenic medium. The rate of mineralization was examined by staining of mineralized nodules with Alizarin red S, and the difference of ALP, BMP-2 and TGF-[β1 between interblocks was examined by enzyme linked immunosorbent assay (ELISA) after 21 d of culture. One way analysis of variance was used for statistical analysis between groups.Results Compared with 5. 5 mmol/L glucose classic group, the rate of mineralization was cut down significantly in 25.0 mmol/L glucose classic group ( (45.3 ± 0.7)% vs ( 68.3 ± 0.8 ) %, P < 0.05 ) , and the activity of ALP (0.350 ± 0.020, 0.563 ± 0.043 ), BMP-2 ( (590 ± 27 ), (744 ± 41 )μg/L) and TGF-β1 ((875 ±40),(1188 ±52)μg/L)were also cut down significantly (all P<0.05). Conclusions High concentration glucose inhibits the differentiation of BMSCs into osteoblastics, which provids a potential mechanisms of diabetes-induced osteoporosis.
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