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目的观察血糖波动和持续性高血糖对小鼠颅骨成骨细胞β-catenin和Cyclin D1基因表达的影响。方法体外培养小鼠颅骨成骨细胞,随机分为3组,正常组(5.5 mmol·L-1)、持续性高糖组(25.0 mmol·L-1)和波动性高糖组(5.5~25.0 mmol·L-1),实时荧光定量聚合酶链反应法检测细胞内β-catenin和Cyclin D1蛋白基因mRNA的转录水平;Western blot法检测β-catenin和Cyclin D1蛋白的表达。结果持续高糖、波动性高糖可显著降低细胞内β-catenin和Cyclin D1基因mRNA的转录水平和蛋白的表达水平,且呈剂量一时间依赖性。结论持续性高糖、波动性高糖可抑制小鼠颅骨成骨细胞细胞增殖,其机制可能是通过抑制β-catenin和Cyclin D1基因表达来实现的。
Objective To observe the effects of blood glucose fluctuation and persistent hyperglycemia on the expression of β-catenin and Cyclin D1 in mouse skull osteoblasts. Methods Calvarial osteoblasts were cultured in vitro and randomly divided into 3 groups: normal control group (5.5 mmol·L-1), continuous high glucose group (25.0 mmol·L-1) and high-glucose group (5.5-25.0 The mRNA levels of β-catenin and Cyclin D1 were detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). The expressions of β-catenin and Cyclin D1 were detected by Western blot. RESULTS: Sustained high glucose and fluctuating high glucose significantly reduced mRNA and protein expression of β-catenin and Cyclin D1 mRNA in a time-dependent manner. CONCLUSION: Sustained hyperglycemia and hyperglycemia can inhibit the proliferation of mouse calvarial osteoblasts. The mechanism may be through inhibiting the expression of β-catenin and Cyclin D1.