摘要
目的:探讨红芪多糖对脂多糖(LPS)诱导的牙周膜细胞凋亡及对Wnt信号通路的影响。方法:体外分离培养人牙周膜细胞。实验分为3组,依次为对照组、脂多糖组和红芪多糖组。对照组细胞用正常的细胞培养液培养细胞;脂多糖组和红芪多糖组细胞用含有脂多糖浓度为10mg/L的培养液培养细胞;红芪多糖组细胞培养液中加入红芪多糖终浓度为10mg/L的培养液。流式细胞仪检测细胞凋亡情况,探针法检测细胞内的活性氧(ROS)水平。试剂盒检测细胞中碱性磷酸酶活性和分泌的肿瘤坏死因子-α(TNF-α)水平。Western blot检测细胞中cmyc、β-连环蛋白(β-catenin)、Bcl-2相关X蛋白(Bax)单克隆抗体、B细胞淋巴瘤/白血病-2(Bcl-2)蛋白水平。结果:脂多糖组细胞存活率明显低于对照组(P<0.01)。脂多糖组细胞凋亡率和Bax水平、ROS水平明显高于对照组(P<0.01)。红芪多糖组细胞凋亡率和Bax水平、ROS水平明显低于脂多糖组(P<0.01)。脂多糖组细胞Bcl-2、c-myc、β-catenin水平和碱性磷酸酶活性明显低于对照组(P<0.01)。红芪多糖组细胞Bcl-2、c-myc、β-catenin水平和碱性磷酸酶活性明显高于脂多糖组(P<0.01)。脂多糖组细胞分泌的TNF-α水平明显高于对照组(P<0.01)。红芪多糖组细胞分泌的TNF-α水平明显低于脂多糖组(P<0.01)。结论:红芪多糖能够抑制脂多糖诱导的人牙周膜细胞凋亡,抑制细胞分泌TNF-α,作用机制可与Wnt信号通路、ROS水平有关。
Objective:To study the effect of Hedysari polysaccharide on apoptosis of periodontal ligament cells induced by LPS and the expression of Wnt signaling pathway.Methods:Human periodontal ligament cells were cultured and divided into three groups,control group,LPS group with normal cell culture medium containing 10μg/mL LPS and Hedysari polysaccharide group with 10μg/mL LPS and 10μg/mL Hedysari polysaccharide.Cell apoptosis was detected by flow cytometry.Intracellular ROS levels were detected by the probe method.The activity of alkaline phosphatase and the levels of TNF-αin the cells were detected by ELISA.The levels of c-myc,β-catenin,Bax and Bcl-2were detected by Western Blot.Results:The apoptosis rate,expression of Bax and ROS level in the LPS group were significantly higher than those in the control group and lower in Hedysari polysaccharide group(P〈0.01).The levels of Bcl-2,c-myc,β-catenin and alkaline phosphatase in the LPS group were significantly lower than those in the control group and significantly higher in Hedysari polysaccharide group(P〈0.01).The level of TNF-αin LPS group was significantly higher than those in the control group and Hedysari polysaccharide group(P〈0.01).Conclusion:Hedysari polysaccharide can inhibit LPS induced apoptosis and the secretion of TNF-αof human periodontal ligament cells.The mechanism may relate to Wnt signaling pathway and ROS level.
出处
《口腔医学研究》
CAS
北大核心
2017年第6期646-649,共4页
Journal of Oral Science Research
基金
河南省医学科技攻关计划项目(编号:201403185)
关键词
人牙周膜细胞
脂多糖
红芪多糖
凋亡
Human periodontal ligament cells
Lipopolysaccharide
Radix Hedysari polysaccharide
Apoptosis