胰岛素对高糖条件下体外培养的大鼠牙囊细胞增殖、分化的影响

Effects of glucose and its modulation by insulin on proliferation and differentiation of rat dental follicle cells in vitro

目的:探讨胰岛素对高糖条件下体外培养的大鼠牙囊细胞(dental follicle cells,DFC_s)增殖、分化的影响。方法:取出生7d的SD大鼠上、下颌磨牙牙囊,原代培养牙囊细胞,选取生长状态良好的第4代细胞,在糖生理浓度(5.5mmol/L)和高糖条件下(16.5 mmol/L和49.5 mmol/L)与6μg/mL胰岛素共同孵育,分别于培养1、3、7、9d时采用MTT法和1～7d采用碱性磷酸酶试剂盒检测胰岛素对高糖条件下体外培养的大鼠牙囊细胞增殖、分化的影响。采用SPSS13.0软件包对数据进行统计学分析。结果:所培养的细胞波形丝蛋白阳性,角蛋白阴性。在第l、3天,16.5mmol/L葡萄糖促进第4代大鼠牙囊细胞的增殖,而49.5 mmol/L葡萄糖抑制牙囊细胞的增殖,胰岛素下调16.5 mmol/L葡萄糖下牙囊细胞的增殖,上调49.5 mmol/L葡萄糖下牙囊细胞增殖;在第7、9天,高糖促进牙囊细胞增殖,胰岛素上调高精下牙囊细胞的增殖。在1～7d内,高糖提高第4代大鼠牙囊细胞碱性磷酸酶活性,胰岛素下调高糖下牙囊细胞的碱性磷酸酶活性(P<0.05)。结论:本实验培养的细胞是来源于外胚层间充质的牙囊细胞。实验结果表明,胰岛素能纠正急性高糖对大鼠牙囊细胞增殖和分化的影响。

PURPOSE： To investigate the effects of glucose and its modulation by insulin on proliferation and differentiation of rat dental follicle cells （DFCs） in vitro. METHODS： The rat dental follicle cells from 7-day neonatal rat were cultured. The effects of different doses of glucose concentration （5.5, 16.5, and 49.5mmol/L）, alone with insulin （61μg/ml）, on rat dental follicle cells was examined in vitro. The DFCs proliferation and alkaline phosphatase （ALP） were examined after 1,3,7,9 days and 1-7 days of culture, respectively. The data were analyzed using SPSS13.0 software package. RESULTS： At 1,3 day, DFCs proliferation greatly enhanced at 16.5mmol/L glucose, and decreased at 49.5mmol/L glucose. Insulin treatment greatly decreased the proliferation of 16.5mmol/L glucose and increased the proliferation of 49.5mmol/L glucose. With high concentrations of glucose, cell proliferation greatly increased at 7,9 day, insulin treatment greatly increased the proliferation of DFCs. High concentrations of glucose greatly increased the DFCs ALP activity in 1-7 days. Insulin treatment greatly decreased DFCs ALP activity with high concentrations of glucose （P〈 0.05）. CONCLUSION： This study suggests that insulin contains the deleterious effect of acute high concentrations of glucose on rat DFCs proliferation and differentiation.