高糖状态下人牙龈上皮细胞的生物学特性

Effects of high glucose on the biological characteristics of human gingival epithelial cells

目的观察高糖状态对人牙龈上皮细胞生物学特性的影响。方法原代培养人牙龈上皮细胞(h GEC),取第三代细胞分为正常组(培养液含5.5 mmol/L D-葡萄糖)、高糖组(培养液含25 mmol/L D-葡萄糖)和甘露醇组(含5.5 mmol/L D-葡萄糖及19.5 mmol/L甘露醇的培养基),采用MTS法检测细胞的增殖情况,流式细胞术检测细胞的生长周期及凋亡情况,划痕实验检测细胞迁移能力。采用重复测量的方差分析进行统计分析。结果高糖组、正常组、甘露醇组三组h GEC相比较,细胞增殖活性(F=5.053,P=0.052)、细胞周期分布(F=1.252,P=0.351)及凋亡细胞比例(F=0.325,P=0.717)差异均无统计学意义;划痕24 h时,糖尿病组的划痕迁移率显著低于正常组(F=54.453,P=0.000)。结论高糖状态下h GEC增殖及凋亡无显著性改变,但细胞迁移能力出现显著下降。

Objective To investigate the biological characteristics of human gingival epithelial cells（ h GECs） in high-glucose environments. Methods Cultured primary h GECs, logarithmic growth phase of the 3rd passage cells were used for subsequent experiments. Experimental groups ：（1） high glucose group： K-SFM medium containing 25 mmol / L D-glucose;（2） normal group： K-SFM medium containing 5.5 mmol / L D-glucose;（3） mannitol group： K-SFM medium containing 5.5 mmol / L Dglucose and 19.5 mmol / L mannitol. We used the MTS method to detect the proliferation of h GECs,applied flow cytometry to detect cell cycle and apoptosis of h GECs, and utilized wound healing test to detect migration of h GECs. Comparisons of the data among different groups were performed by repeated measures analysis of variance（ANOVA）. Results There is no statistically significant difference in cell proliferation activity（F = 5.053,P = 0.052）, proportion of cell proliferate（DNA synthesis phase）（F =1.252,P = 0.351） and cell apoptosis（F = 0.325,P = 0.717）. Wound healing test showed after culture for24 h, the scratch migration rate of h GECs in the high group was significantly lower than cells in the normal glucose group and mannitol group（F = 54.453,P = 0.000）. Conclusion High glucose had no significantly effect on cell morphology, cell proliferation and apoptosis of h GECs. However, it weakened the ability of cell migration of hGECs.