中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2014年
9期
535-539
,共5页
于雅琼%仇丽鸿%郭佳杰%曲柳%徐莉雅%钟鸣
于雅瓊%仇麗鴻%郭佳傑%麯柳%徐莉雅%鐘鳴
우아경%구려홍%곽가걸%곡류%서리아%종명
卟啉单胞菌,牙髓%脂多糖类%成骨细胞%巨噬细胞集落刺激因子%NF-κB
卟啉單胞菌,牙髓%脂多糖類%成骨細胞%巨噬細胞集落刺激因子%NF-κB
계람단포균,아수%지다당류%성골세포%거서세포집락자격인자%NF-κB
Porphyromonas endodontalis%Lipopolysaeeharides%Osteoblasts%Macrophage colony-stimulating factor%NF-kappa B
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.
目的 探討牙髓卟啉單胞菌(Porphyromonas endodontalis,Pe)脂多糖對成骨細胞產生巨噬細胞集落刺激因子(macrophage colony stimulating factor,M-CSF) mRNA和蛋白錶達的影響及是否有覈因子κB信號通路的參與.方法 以不同質量濃度的Pe-脂多糖(0～50 mg/L)刺激MC3T3-E1細胞和以10 mg/L Pe-脂多糖作用于細胞不同時間(0～ 24 h)後,採用反轉錄-PCR和酶聯免疫吸附試驗(enzyme-linked immunoadsordent assay,ELISA)檢測M-CSF mRNA和蛋白的錶達,其中未加Pe-脂多糖刺激組為空白對照組.採用覈因子κB信號通路特異性抑製劑BAY 11-7082預處理1h,檢測其對Pe-脂多糖刺激MC3T3-E1細胞後M-CSF mRNA和蛋白錶達的影響,實驗分組如下:空白對照組、BAY組(10 μmol/L BAY 11-7082單獨作用MC3T3-E1細胞)、Pe-脂多糖組(10 mg/L的Pe-脂多糖刺激MC3T3-E1細胞6h)、BAY+Pe-脂多糖組(10 μmol/L BAY 11-7082預處理細胞1h,10 mg/L的Pe-脂多糖刺激MC3T3-E1細胞6h).結果 不同質量濃度Pe-脂多糖(0～50 mg/L)刺激MC3T3-E1細胞後,M-CSFmRNA和蛋白的錶達具有劑量依賴性,蛋白錶達量從(35±2) ng/L(空白對照組)增加到(170±8) ng/L(50 mg/L組).噹10 mg/L Pe-脂多糖作用于MC3T3-E1細胞6h時,M-CSF mRNA的錶達量最大,隨著作用時間的延長,M-CSF mRNA錶達量逐漸下降;噹10 mg/L Pe-脂多糖作用于MC3T3-E1細胞10 h時M-CSF的蛋白錶達量為(122±4) ng/L,隨著作用時間延長,M-CSF mRNA的蛋白錶達量逐漸下降.10 μmol/L的BAY 11-7082預處理細胞1h後,顯著降低瞭Pe-脂多糖誘導的M-CSF mRNA和蛋白的錶達水平,BAY組與空白對照組相比差異無統計學意義.結論 Pe-脂多糖可能通過激活覈因子κB信號通路誘導成骨細胞錶達M-CSF mRNA和蛋白.
목적 탐토아수계람단포균(Porphyromonas endodontalis,Pe)지다당대성골세포산생거서세포집락자격인자(macrophage colony stimulating factor,M-CSF) mRNA화단백표체적영향급시부유핵인자κB신호통로적삼여.방법 이불동질량농도적Pe-지다당(0～50 mg/L)자격MC3T3-E1세포화이10 mg/L Pe-지다당작용우세포불동시간(0～ 24 h)후,채용반전록-PCR화매련면역흡부시험(enzyme-linked immunoadsordent assay,ELISA)검측M-CSF mRNA화단백적표체,기중미가Pe-지다당자격조위공백대조조.채용핵인자κB신호통로특이성억제제BAY 11-7082예처리1h,검측기대Pe-지다당자격MC3T3-E1세포후M-CSF mRNA화단백표체적영향,실험분조여하:공백대조조、BAY조(10 μmol/L BAY 11-7082단독작용MC3T3-E1세포)、Pe-지다당조(10 mg/L적Pe-지다당자격MC3T3-E1세포6h)、BAY+Pe-지다당조(10 μmol/L BAY 11-7082예처리세포1h,10 mg/L적Pe-지다당자격MC3T3-E1세포6h).결과 불동질량농도Pe-지다당(0～50 mg/L)자격MC3T3-E1세포후,M-CSFmRNA화단백적표체구유제량의뢰성,단백표체량종(35±2) ng/L(공백대조조)증가도(170±8) ng/L(50 mg/L조).당10 mg/L Pe-지다당작용우MC3T3-E1세포6h시,M-CSF mRNA적표체량최대,수착작용시간적연장,M-CSF mRNA표체량축점하강;당10 mg/L Pe-지다당작용우MC3T3-E1세포10 h시M-CSF적단백표체량위(122±4) ng/L,수착작용시간연장,M-CSF mRNA적단백표체량축점하강.10 μmol/L적BAY 11-7082예처리세포1h후,현저강저료Pe-지다당유도적M-CSF mRNA화단백적표체수평,BAY조여공백대조조상비차이무통계학의의.결론 Pe-지다당가능통과격활핵인자κB신호통로유도성골세포표체M-CSF mRNA화단백.
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.
