中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2012年
z1期
145-148
,共4页
牙周膜%成纤维细胞%应力,物理%转录因子%黏着斑激酶
牙週膜%成纖維細胞%應力,物理%轉錄因子%黏著斑激酶
아주막%성섬유세포%응력,물리%전록인자%점착반격매
Periodontal ligament%Fibroblasts%Stress,mechanical%Transcription factors%Focal adhesion kinase
目的 研究动态张应力刺激下体外培养的人牙周膜成纤维细胞(periodontal ligament fibroblasts,PDLF)转录因子c-fos和c-jun mRNA的变化以及黏着斑激酶(focal adhesion kinase,FAK)对c-fos和c-jun mRNA表达的影响,为进一步探索人PDLF中力学信号转导途径提供依据.方法 刮取牙根中1/3牙周膜组织,采用Ⅰ型胶原酶消化结合组织块贴附法进行PDLF原代培养并传代.用四点弯曲加载装置对体外培养的第六代PDLF分别施加动态张应力,以加入FAK抗体作为抗体组,不加入FAK抗体作为对照组,采用实时荧光定量PCR法研究动态张应力下FAK对人PDLF转录因子c-fos和c-jun mRNA表达的影响.对实时荧光定量PCR所得数据取常用对数后行单因素方差分析.结果 抗体组加力0、0.5、1、2、4h后c-fos mRNA表达量分别为(1.64±0.20)、(1.67±0.28)、(1.57±0.18)、(1.44±0.26)、(1.33±0.29) lg拷贝数/μg；c-jun mRNA表达量分别为(1.56±0.67)、(1.60±0.21)、(1.46±0.31)、(1.41±0.13)、(1.38±0.14) lg拷贝数/μg.c-fos和c-jun mRNA表达量在加力后上调,0.5h达到峰值,0.5h后出现持续性下调.抗体组细胞c-fos和c-jun mRNA表达变化规律与对照组相似,但同一时段抗体组较相应对照组高,1h后差异有统计学意义(P＜0.05).结论 转录因子c-fos和c-jun可能在牙周膜对机械力刺激的反应中发挥作用,FAK可能是人PDLF中力学信号转导途径的重要分子.
目的 研究動態張應力刺激下體外培養的人牙週膜成纖維細胞(periodontal ligament fibroblasts,PDLF)轉錄因子c-fos和c-jun mRNA的變化以及黏著斑激酶(focal adhesion kinase,FAK)對c-fos和c-jun mRNA錶達的影響,為進一步探索人PDLF中力學信號轉導途徑提供依據.方法 颳取牙根中1/3牙週膜組織,採用Ⅰ型膠原酶消化結閤組織塊貼附法進行PDLF原代培養併傳代.用四點彎麯加載裝置對體外培養的第六代PDLF分彆施加動態張應力,以加入FAK抗體作為抗體組,不加入FAK抗體作為對照組,採用實時熒光定量PCR法研究動態張應力下FAK對人PDLF轉錄因子c-fos和c-jun mRNA錶達的影響.對實時熒光定量PCR所得數據取常用對數後行單因素方差分析.結果 抗體組加力0、0.5、1、2、4h後c-fos mRNA錶達量分彆為(1.64±0.20)、(1.67±0.28)、(1.57±0.18)、(1.44±0.26)、(1.33±0.29) lg拷貝數/μg；c-jun mRNA錶達量分彆為(1.56±0.67)、(1.60±0.21)、(1.46±0.31)、(1.41±0.13)、(1.38±0.14) lg拷貝數/μg.c-fos和c-jun mRNA錶達量在加力後上調,0.5h達到峰值,0.5h後齣現持續性下調.抗體組細胞c-fos和c-jun mRNA錶達變化規律與對照組相似,但同一時段抗體組較相應對照組高,1h後差異有統計學意義(P＜0.05).結論 轉錄因子c-fos和c-jun可能在牙週膜對機械力刺激的反應中髮揮作用,FAK可能是人PDLF中力學信號轉導途徑的重要分子.
목적 연구동태장응력자격하체외배양적인아주막성섬유세포(periodontal ligament fibroblasts,PDLF)전록인자c-fos화c-jun mRNA적변화이급점착반격매(focal adhesion kinase,FAK)대c-fos화c-jun mRNA표체적영향,위진일보탐색인PDLF중역학신호전도도경제공의거.방법 괄취아근중1/3아주막조직,채용Ⅰ형효원매소화결합조직괴첩부법진행PDLF원대배양병전대.용사점만곡가재장치대체외배양적제륙대PDLF분별시가동태장응력,이가입FAK항체작위항체조,불가입FAK항체작위대조조,채용실시형광정량PCR법연구동태장응력하FAK대인PDLF전록인자c-fos화c-jun mRNA표체적영향.대실시형광정량PCR소득수거취상용대수후행단인소방차분석.결과 항체조가력0、0.5、1、2、4h후c-fos mRNA표체량분별위(1.64±0.20)、(1.67±0.28)、(1.57±0.18)、(1.44±0.26)、(1.33±0.29) lg고패수/μg；c-jun mRNA표체량분별위(1.56±0.67)、(1.60±0.21)、(1.46±0.31)、(1.41±0.13)、(1.38±0.14) lg고패수/μg.c-fos화c-jun mRNA표체량재가력후상조,0.5h체도봉치,0.5h후출현지속성하조.항체조세포c-fos화c-jun mRNA표체변화규률여대조조상사,단동일시단항체조교상응대조조고,1h후차이유통계학의의(P＜0.05).결론 전록인자c-fos화c-jun가능재아주막대궤계력자격적반응중발휘작용,FAK가능시인PDLF중역학신호전도도경적중요분자.
Objectives To investigate the effect of focal adhesion kinase (FAK) on the mRNA expression of transcription factors c-fos/c-jun in cultured human periodontal ligament fibroblasts (hPDLF)loaded with tensional force.Methods The periodontal ligament,which was attached to the mid-third part of the fresh root of young premolars extracted for orthodontic purposes,was removed.The hPDLF was cultured using the method of digesting by Ⅰ-type collagenase combined with tissue adhering.Then hPDLF was isolated and purified by cells passage and PDLF were used in the experiments at passage six.A four-point bending device named " Forcel" was used for force loading.Cells cultured in vitro were loaded with 2000 μstrain tensional force and collected at different time (0,0.5 h,1 h,2 h,or 4 h) after strain loading.Cells were divided into two groups (with or without FAK antibody).Aherations in the mRNA expression of transcription factors c-fos/c-jun due to cyclic tensional force were investigated using real-time polymerase chain reaction (PCR).ANOVA and LSD were used to process the data obtained.Results After strain loading the c-fos/c-jun mRNA expression was upregulated,with a peak at 0.5 h,and then downregulated after 0.5 h.The levels of gene expression in cells treated with the FAK antibody were greater than those in cells untreated with the FAK antibody (P ＜0.05).Conclusions These results provided evidence that transcription fac tors c-fos/c-jun probably played a critical part in the response of periodontal tissue (periodontal ligament fibroblasts) to mechanical stimulation and that FAK probably played an important role in the mechanical signaling pathway of hPDLF.
