摘要
目的 探讨牙髓卟啉单胞菌（Porphyromonas endodontalis,Pe）脂多糖对成骨细胞产生巨噬细胞集落刺激因子（macrophage colony stimulating factor,M-CSF） mRNA和蛋白表达的影响及是否有核因子κB信号通路的参与.方法 以不同质量浓度的Pe-脂多糖（0～50 mg/L）刺激MC3T3-E1细胞和以10 mg/L Pe-脂多糖作用于细胞不同时间（0～ 24 h）后,采用反转录-PCR和酶联免疫吸附试验（enzyme-linked immunoadsordent assay,ELISA）检测M-CSF mRNA和蛋白的表达,其中未加Pe-脂多糖刺激组为空白对照组.采用核因子κB信号通路特异性抑制剂BAY 11-7082预处理1h,检测其对Pe-脂多糖刺激MC3T3-E1细胞后M-CSF mRNA和蛋白表达的影响,实验分组如下：空白对照组、BAY组（10 μmol/L BAY 11-7082单独作用MC3T3-E1细胞）、Pe-脂多糖组（10 mg/L的Pe-脂多糖刺激MC3T3-E1细胞6h）、BAY＋Pe-脂多糖组（10 μmol/L BAY 11-7082预处理细胞1h,10 mg/L的Pe-脂多糖刺激MC3T3-E1细胞6h）.结果 不同质量浓度Pe-脂多糖（0～50 mg/L）刺激MC3T3-E1细胞后,M-CSFmRNA和蛋白的表达具有剂量依赖性,蛋白表达量从（35±2） ng/L（空白对照组）增加到（170±8） ng/L（50 mg/L组）.当10 mg/L Pe-脂多糖作用于MC3T3-E1细胞6h时,M-CSF mRNA的表达量最大,随着作用时间的延长,M-CSF mRNA表达量逐渐下降;当10 mg/L Pe-脂多糖作用于MC3T3-E1细胞10 h时M-CSF的蛋白表达量为（122±4） ng/L,随着作用时间延长,M-CSF mRNA的蛋白表达量逐渐下降.10 μmol/L的BAY 11-7082预处理细胞1h后,显著降低了Pe-脂多糖诱导的M-CSF mRNA和蛋白的表达水平,BAY组与空白对照组相比差异无统计学意义.结论 Pe-脂多糖可能通过激活核因子κB信号通路诱导成骨细胞表达M-CSF mRNA和蛋白.
Objective To investigate the effects of lipopolysaccharides（LPS） extracted from Porphyromonas endodontalis（Pe） on the expression of macrophage colony stimulating factor（M-CSF） mRNA and protein in MC3T3-E1 cells and the role of nucler factor-κB（NF-κB） in the process.Methods MC3T3-E1 cells were treated with different concentrations of Pe-LPS（0-50 mg/L） and 10 mg/L Pe-LPS for different hours （0-24 h）.The expression of M-CSF mRNA and protein was detected by reverse transcription polymerase chain reaction（RT-PCR） and enzyme-linked immunoadsordent assay（ELISA）.The cells untreated by Pe-LPS served as control.The expression of M-CSF mRNA and protein was also detected in 10 mg/L Pe-LPS treated MC3T3-E1 cells after pretreated with BAY 11-7082 for 1 h,a special NF-κB inhibitor.The groups were divided as follows,control group,BAY group（10 μmol/L BAY 11-7082 treated alone MC3T3-E1 cells）,Pe-LPS group（10 mg/L Pe-LPS stimulated MC3T3-E1 cells for 6 h）,BAY combine with Pe-LPS group （10 μmol/L BAY 11-7082 pretreated cells for 1 h and 10 mg/L of Pe-LPS stimulated MC3T3-E1 cells for 6 h）.Results The level of M-CSF mRNA and protein increased significantly after treatment with different concentrations of Pe-LPS（0-50 mg/L）,which indicated that Pe-LPS induced osteoblasts to express M-CSF mRNA and protein in dose dependent manners.The expression of M-CSF protein increased from（35±2） ng/L （control group） to （170±8） ng/L（50 mg/L group）.Maximal induction of M-CSF mRNA expression was found in the MC3T3-E1 cells treated with 10 mg/L Pe-LPS for 6 h.After 6 h,the expression of M-CSF mRNA decreased gradually.The expression of M-CSF protein also increased with the treatment of 10 mg/L Pe-LPS for 10 h[（122±4） ng/L].After 10 h,the expression of M-CSF protein decreased gradually.The mRNA and proteins of M-CSF decreased significantly after pretreatment with 10 μmol/L BAY 11-7082 for 1 h.There was no significant difference between BAY group and the control.Conclusions Pe-LPS may induce the expression of M-CSF mRNA and protein in MC3T3-E 1 cells through the signaling of NF-κB.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2014年第9期535-539,共5页
Chinese Journal of Stomatology
基金
辽宁省科学技术计划（2011225020）
