中文摘要：

目的 观察高糖环境对大鼠下颌骨骨髓基质细胞增殖的影响,并结合Wnt/β-链蛋白信号通路初步探讨高糖对骨髓基质细胞增殖的影响.方法 从Wistar大鼠下颌骨中分离培养骨髓基质细胞,分别给予不同糖浓度（5.5和16.5 mmol/L）干预,采用噻唑蓝法观察骨髓基质细胞1、3、5、7d时的增殖情况,采用流式细胞仪对高糖干预5d细胞进行细胞周期检测,Western免疫印迹检测5d时β-链蛋白和细胞周期蛋白D1的表达,实时定量多聚酶链反应检测淋巴增强因子-1和细胞周期蛋白D1的mRNA表达.结果 噻唑蓝法检测结果显示,1d骨髓基质细胞在两种糖浓度中的光密度值差异无统计学意义（P=0.700）,3、5、7d时16.5 mmol/L组的细胞光密度值显著低于5.5 mmol/L组（P=0.006,P=0.002,P=0.003）.5d的细胞周期结果显示,与5.5 mmol/L组细胞相比,16.5 mmol/L组细胞可以抑制细胞从G1期向S期转化,16.5 mmol/L组细胞的增殖指数显著低于5.5 mmol/L组（P=0.014）.与5.5 mmol/L组相比,16.5 mmol/L组细胞明显抑制β-链蛋白的表达,同时Wnt/β-链蛋白通路中淋巴增强因子-1 mRNA及下游细胞周期蛋白D1 mRNA和蛋白的表达也受到明显抑制.结论 高糖环境可以通过抑制Wnt/β-链蛋白信号通路中关键因子β-链蛋白、淋巴增强因子-1及下游与细胞增殖相关靶基因细胞周期蛋白D1的表达,进而抑制骨髓基质细胞的增殖.

英文摘要：

Objective To explore the effect of high glucose on proliferation of bone marrow stromal stem cells through Wnt/β-catenin pathway.Methods Bone marrow stormal cells were obtained from the mandible of Wistar rats and stimulated with different concentrations of glucose （5.5 and 16.5 mmol/L）.Cell proliferation was evaluated with methyl thiazolyl tetrazolium assay （1,3,5,and 7 d） and cell cycle analysis by flow cytometry （5 d）.β-catenin and cyclin D1 protein levels were determined by Western blot.The mRNA expression of lymphoid enhancer binding factor-1 （LEF-1） and cyclin D1 were tested by real-time polymerase chain reaction.Results The results of methyl thiazolyl tetrazolium assay indicated that the optical density values of two different concentrations of the glucose had no statistical difference on day 1 （P =0.700）.On days 3,5,and 7,the optical density values of the 16.5 mmol/L group were significantly lower than those in the 5.5 mmol/L group （P =0.006,P =0.002,and P =0.003）.Cell cycle analysis indicated that high glucose concentration could reduced the progression from phase G1 to S,and the proliferation index values of the 16.5 mmol/L group were significantly lower than those of the 5.5 mmol/L group （P =0.014）.The β-catenin and cyclin D1 levels were lower in the 16.5 mmol/L group when compared with the 5.5 mmol/L group.High glucose condition also reduced the mRNA expressions of LEF-1 and cyclin D1.Conclusion High glucose can inhibit the proliferation of bone marrow stormal cells by suppressing the expressions of β-catenin,LEF-1,and cyclin D1 in the Wnt/β-catenin pathway.