骨化三醇对不同维生素D受体FokⅠ基因型人牙周膜细胞增殖的抑制作用

The inhibitory effect of calcitriol on the proliferation of hPDLCs populations of different vitamin D receptor Fok Ⅰ genotypes

目的:观察骨化三醇[1,25-(OH)2D3,VD3]对人牙周膜细胞(human periodontal ligament cells,hPDLCs)增殖的作用并分析维生素D受体FokⅠ多态性位点(rs2228570,T/C)对此作用的影响。方法:原代培养个体来源hPDLCs群,限制性内切酶片段长度多态性法检测细胞群维生素D受体FokⅠ多态性位点基因型。选取第4代具有FF、Ff、ff基因型hPDLCs群,用0.1%无水乙醇(对照组)、10-14～10-8 mol/L VD3分别处理1d、3d、5d、7d,MTT法检测细胞的增殖情况。用SPSS11.5软件包对数据进行单因素方差分析和LSD Post Hoc检验。结果:VD3以时间和剂量依赖性抑制3种基因型hPDLCs群的增殖。与对照组和其他各浓度组相比,10-8mol/L VD3的抑制作用最强,第5天或第7天达到最大抑制效应。与相应的对照组比较,对FF细胞群产生显著抑制效应的最低VD3浓度为10-12mol/L(P<0.01),Ff为10-10mol/L(P<0.01),ff为10-8 mol/L(P<0.01)。10-8 mol/L VD3对ff细胞群增殖的最大抑制率为16.93%,显著低于FF(31.20%,P<0.05)和Ff(29.68%,P<0.05)细胞群。第3天和第5天,10-8mol/L VD3对不同基因型细胞群增殖产生的抑制作用由强至弱为FF>Ff>ff(P<0.01)。结论:VD3能够显著抑制hPDLCs的增殖,维生素D受体FokⅠ多态性位点可能对这一过程产生影响。

PURPOSE：The study was designed to investigate the effect of calcitriol [1,25-（OH）2D3,VD3] on the proliferation of human periodontal ligament cells（hPDLCs） and to analyze the influence of vitamin D receptor FokⅠ polymorphism（rs2228570,T/C） on the effect.METHODS：Primary cultures of hPDLCs populations from individual donors were established.Restriction fragment length polymorphism method was used to detect the genotypes of the FokⅠ site.The fourth passage hPDLCs populations of FF,Ff and ff genotypes were respectively treated with 0.1% ethyl alcohol（control） or 10-14-10-8 mol/L VD3 for 1,3,5 and 7 days.MTT method was performed to evaluate the proliferation of hPDLCs.The data was analyzed by one-way ANOVA and LSD Post Hoc test using SPSS 11.5 software package.RESULTS：VD3 inhibited the proliferation of hPDLCs populations of three genotypes in a time-and concentration-dependent manner.10-8 mol/L VD3 exerted the most powerful inhibition compared with controls and other concentrations and the highest inhibition occurred on day 5 or day 7.Compared with the corresponding controls,the lowest concentrations of VD3 which exerted statistically significant inhibitory effect on the proliferation of hPDLCs populations were listed as follows：10-12mol/L for the hPDLCs population of FF genotype（P〈0.01）,10-10 for Ff genotype（P〈0.01） and 10-8 mol/L for ff genotype（P〈0.01）.The highest proliferation inhibitory rate exerted by 10-8 mol/L VD3 on the ff population was 16.93%,which was significantly lower than FF（31.20%,P〈0.05） and Ff（29.68%,P〈0.05）.On day 3 and day 5,10-8 mol/L VD3exhibited the strongest suppression on the proliferation of hPDLCs population of FF genotype,followed by Ff and ff genotypes（P〈0.01）.CONCLUSION：VD3 can significantly inhibit the proliferation of hPDLCs,which may be affected by VDR FokⅠ polymorphism.