胰岛素对大鼠牙周膜细胞增殖及其炎性因子表达的影响

Effect of Insulin on Proliferation and Expression of Inflammatory Factors of Rat Periodontal Ligament Cells

目的:研究胰岛素对大鼠牙周膜细胞增殖及其炎性因子表达的影响。方法:分离培养原代大鼠牙周膜细胞,在体外高糖培养液培养,分别加入浓度为0nmol/L、25nmol/L、50nmol/L和100nmol/L的胰岛素,0nmol/L浓度为对照组,其余均为实验组,用不同浓度的胰岛素处理细胞后,MTT法检测大鼠牙周膜细胞增殖水平变化;根据MTT实验结果,选取25nmol/L浓度胰岛素处理组为实验组,进行炎性因子表达差异研究,实时定量RT-PCR检测不同浓度胰岛素处理后炎性因子IL-1β、PGE2、TNF-α、IFN-γ、IL-10的表达;ELISA法检测IL-1β、PGE2、TNF-α、IFN-γ、IL-10含量;通过上述实验验证胰岛素对牙周膜细胞增殖及对牙周膜细胞炎性因子表达的影响。结果:与对照组相比,随着胰岛素浓度的增高,抑制作用越明显;在mRNA水平和蛋白水平,胰岛素处理的实验组牙周膜细胞,其炎性因子表达量均明显增高(P<0.05)。结论:胰岛素会抑制大鼠牙周膜细胞的增殖,且对牙周膜细胞的炎性因子表达有促进作用。

Objective To study the effect of insulin on the proliferation of rat periodontal ligament cells and the expression of in flammatory factors.Methods Primary rat periodontal ligament cells were isolated and cultured in high glucose medium in vitro.The concentration of insulin was 0nmol/L,25nmol/L,50nmol/L and 100nmol/L.The control group was 0nmol/L.The rest were experimental groups.After treated with different concentrations of insulin,the proliferation level of rat periodontal ligament cells was detected by MTT assay.According to MTT test results,25nmol/L insulin treatment group was selected as the experimental group,the difference of expression of inflammatory factors was studied.After treated with different concentrations of insulin,the proliferation level of rat periodontal ligament cells was detected by MTT test,and the expression of inflammatory factors IL-1β,PGE2,TNF-α,IFN-γand IL-10 was detected by real-time quantitative RT-PCR.The contents of IL-1β,PGE2,TNF-α,IFN-γand IL-10 were measured by ELISA.The effects of insulin on the proliferation of periodontal ligament cells and the expression of inflammatory factors in periodontal ligament cells were veri fied by the above experiments.Results Compared with the control group,the inhibition was more obvious with the increase of insulin concentration,and the expression of inflammatory factors in periodontal ligament cells treated with insulin increased signi ficantly at the level of protein and mRNA(P<0.05).Conclusion Insulin can inhibit the proliferation of rat periodontal ligament cells and promote the expression of inflammatory factors in periodontal ligament cells.