不同静压力下脂多糖刺激牙周膜成纤维细胞对炎症相关因子表达的影响

目的:探讨牙周炎中牙龈卟啉单胞菌LPS(Porphyromonas gingivalis Lipopolysaccharide,Pg.LPS)和人牙周膜成纤维细胞(human periodontal ligament fibroblast,HPDLF)与正畸炎症相关的牙根吸收作用机制。方法:取4~6代体外培养的HPDLF,MTT法检测1~10μg/ml Pg.LPS对HPDLF增殖活性的影响,再选择最佳浓度作用于HPDLF,加载0~5g/cm^2静压力24h,实验组分为静压力组和Pg.LPS+静压力组,通过qRT-PCR和ELISA检测HPDLF白细胞介素17(interleukin 17,IL-17)及白细胞介素6(interleukin 6,IL-6)的mRNA和蛋白表达水平。结果:①HPDLF增殖在1~4μg/ml Pg.LPS刺激时随浓度增加而增加,在5~7μg/ml随浓度增加而减少,8~10μg/ml细胞增殖明显受到抑制;②两组中IL-17、IL-6的表达量随静压力值增加而增强,相同力值下对照组的IL-17、IL-6表达量高于实验组,差异有统计学意义。结论:Pg.LPS浓度过高能抑制HPDLF增殖,而适当的浓度则促进其增殖;Pg.LPS和静压力均能刺激HPDLF产生炎症相关因子,参与正畸炎症相关牙根吸收发生发展的过程。

Effects of Lipopolysaccharides on the Expression of Proinflammatory Factors in Periodontal Fibroblasts Stimulated by Lipopolysaccharides under Different Static Pressure

Objective To investigate the mechanism of Porphyromonas gingivalis Lipopolysaccharide (Pg.LPS) and human periodontal fibroblast (HPDLF) in orthodontic inflammatory-related root resorption in periodontitis.Methods The HPDLF cultured in vitro for 4-6 generations was used to detect the effect of 1-10 μg/ml Pg.LPS on the proliferation of HPDLF by MTT assay.Then choose the optimal concentration stimulate HPDLF and load 0-5 g/cm^2 static pressure strength for 24 hours.The experimental group was divided into static pressure group and Pg.LPS + static pressure group.QRT-PCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of interleukin-17 and interleukin-6 in HPDLF.Results ①HPDLF proliferation increased with the increase of concentration in 1-4 μg/ml Pg.LPS stimulation,decreased with the increase of concentration in 5-7 μg/ml,and was significantly inhibited cell proliferation in 8-10 μg/ml.② The expressions of interleukin 17 and interleukin 6 in the static pressure group and Pg.LPS + static pressure group increased with the increase of static pressure value,and the expressions of interleukin 17 and interleukin 6 in the static pressure group were higher than those in Pg.LPS + static pressure group under the same force value,and the difference was statistically significant.Conclusion Excessive concentration of Pg.LPS inhibited the proliferation of HPDLF,and the appropriate concentration could promote the proliferation of HPDLF.Both Pg.LPS and static pressure can stimulate HPDLF to produce pro-inflammatory factors and participate in the development of orthodontic inflammatory-related root resorption,but Pg.LPS inhibits the secretion of pro-inflammatory factors by static pressure on HPDLF.