压力及粪肠球菌上清液对人牙周膜细胞骨吸收相关因子表达的影响研究

The effects of Enterococcus faecalis supernatant on the expression of bone resorption related factors in human periodontal ligament cells under pressure

目的:探讨压力及粪肠球菌上清液对人牙周膜细胞(hPDLCs)骨吸收相关因子表达的影响。方法:CCK-8法检测不同浓度的粪肠球菌上清液对hPDLCs增殖活性的影响,倒置显微镜观察不同压力大小引发hPDLCs的细胞形态变化;将hPDLCs分成压力组、压力+粪肠球菌组,每组均分别加载0、49、196和392 Pa压力;通过RT-PCR和ELISA检测hPDLCs RANKL及OPG的mRNA和可溶性蛋白表达水平。结果:10%的粪肠球菌上清液能够降低hPDLCs增殖活性(P<0.05);RT-PCR结果显示,细胞在受压力≥196 Pa时,RANKL mRNA表达增加,OPG mRNA表达减小(P<0.05),在有细菌上清液存在情况下趋势更明显。蛋白表达趋势与mRNA基本一致。结论:过度的机械压力可引起hPDLCs骨吸收因子的高表达,粪肠球菌上清液可进一步加重这种作用。

Objective:To investigate the effects of pressure and Enterococcus faecalis(E.faecalis)supernatant on the expression of bone resorption related factors in human periodontal ligament cells(hPDLCs).Methods:HPDLCs were divided into pressure groups and pressure+E.faecalis groups.The cells in the groups were loaded with 0,49,196 and 392 Pa pressure respectively.CCK-8 assay was used to detect the effect of different concentrations of E.faecalis supernatant on the proliferation of hPDLCs,and the morphological changes of hPDLCs induced by different pressure was observed by inverted microscope.The mRNA and soluble protein expression levels of RANKL and OPG were detected by RT-PCR and ELISA respectively.Results:10%E.faecalis supernatant reduced the proliferation of hPDLCs(P<0.05).RT-PCR showed that when the cells were under the pressure≥196 Pa,the mRNA expression of RANKL increased and that of OPG decreased(P<0.05),and the trend was more obvious in the presence of bacterial supernatant.The trend of protein expression was basically consistent with that of mRNA.Conclusion:Excessive mechanical pressure can cause high expression of bone resorption factors of hPDLCs,which can be further aggravated by E.faecalis supernatant.