脂多糖对人牙周膜干细胞增殖及炎症因子表达的影响

摘要

目的:探讨脂多糖(LPS)对人牙周膜干细胞(hPDLSCs)增殖及炎症因子表达水平的影响.方法:体外分离培养hPDLSCs,并将其随机分为A、B、C组,A、B组分别用含LPS终浓度为10 μg/mL和10 ng/mL的α-MEM进行培养,C组用不含LPS的α-MEM进行培养作为对照;分别于培养12、24、48、72 h后,用MTT法检测各组细胞的增殖情况,同时取培养72 h后的各组细胞,用RT-PCR检测其IL-6、IL-1 β、TNF-αmRNA的表达水平.结果:MTT检测结果显示,各组在各时间点的A490值由高到底依次为:B组＞C组＞A组,3组间两两相比均有统计学差异(P＜ 0.05);RT-PCR检测结果显示,与C组相比,A、B组的IL-6、IL-1β、TNF-α mRNA表达水平均显著升高,且以A组升高更明显,3组间两两相比均有统计学差异(P＜0.05).结论:低浓度LPS对hPDLSCs的增殖具有一定的促进作用,而高浓度的LPS对hPDLSCs的增殖则具有一定的抑制作用,并能促进炎症因子的表达.%AIM:To investigate the effects of lipopolysaccharide (LPS) on the proliferation and expression of inflammatory cytokines in human periodontal ligament stem cells (hPDLSCs).METHODS:hPDLSCs were isolated,cultured and identified from 18 extracted healthy premolars for orthodontic reasons.hPDLSCs were stimulated with LPS at 10 μg/mL and 10 ng/mL respectively and cultured in MEM medium for 72 h.Cells without LPS stimulation served as the controls.Cell proliferation was examined by MTT assay at 12,24 and 72 h of culture.mRNA were extracted at the end of 72 h culture and were subjected to RT-PCR for the determination of IL-6,IL-1β and TNF-α expression.RESULTS:10 ng/mL LPS increased the proliferation of hPDLSCs while 10 μg/mL LPS inhibited the proliferation (P ＜ 0.05).LPS dose-dependently increased IL-6,IL-1 β and TNF-α expression in hPDLSCs (P ＜ 0.05).CONCLUSION:Low concentration of LPS stimulates hPDLSCs growth but high concentration of LPS inhibits hPDLSCs proliferation.Furthermore,LPS may increase inflammatory cytokines expression in hPDLSCs.