马甲子对LPS刺激下人牙髓细胞炎性因子表达影响的初探

Effects of Paliurus Ramosissimus on the Expression of Inflammatory Factors in Human Dental Pulp Cells Stimulated by LPS

目的:研究马甲子提取物对体外培养HDPCs生长及对P.g-LPS诱导HDPCs合成分泌的炎症相关细胞因子IL-1β、IL-6及TNF-α的影响。方法:体外培养人牙髓细胞,建立炎症状态的体外HDPCs,检测不同浓度梯度马甲子提取物对HDPCs生长的影响,以及对炎症状态下的HDPCs各炎性因子mRNA表达及蛋白表达的影响。结果:经P.g-LPS刺激HDPCs 3 h,可成功形成体外HDPCs的炎症状态。在本实验条件下,125μg·mL^(-1)的马甲子提取物溶液可以保证HDPCs的生长,同时可以下调炎症环境下HDPCs的各炎性因子的表达。结论:马甲子提取物能对牙髓细胞炎症反应产生一定抑制作用,为马甲子在牙髓炎症治疗中的应用提供了实验依据。

Objective:Here,we investigated the effects of Paliurus Ramosissimus extracts on the proliferation and inflammatory factors(IL-1β,IL-6 and TNF-α)expression of human dental pulp cells(HDPCs).Methods:HDPCs were cultured and stimulated by P.g LPS to establish the inflammatory cell model.The mRNA expression of inflammatory cytokines(IL-1β,IL-6 and TNF-α)were analyzed by RT-PCR.The proliferation and the inflammatory cytokines expression of HDPCs were detected by CCK-8 analysis and RT-PCR,respectively.ELISA was used to detect the protein expression changes of inflammatory cytokines.Results:Paliurus Ramosissims extracts at 125μg·mL^(-1) was the appropriate concentration without impairments of cell proliferation(P<0.05).The mRNA expression of inflammatory cytokines were significantly up-regulated and reached the peak at 3 hours after P.g LPS stimulation(P<0.05).It was found that the extracts of Paliurus Ramosissims can markedly reduce the expression of inflammatory cytokines(IL-1β,IL-6,TNF-α)of the HDPCs in the inflammatory environment(P<0.05).Conclusion:The inflammatory model of HDPCs in vitro can be successfully established by stimulating HDPCs with P.g-LPS for 3 hours.Under the condition of this experiment,the extracts of Paliurus Ramosissims had certain cytocompatibility,can ensure the growth of the HDPCs,and can also down-regulate the expression of the inflammatory cytokines(IL-1β,IL-6,TNF-α)of the HDPCs in the inflammatory environment.