中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2010年
2期
80-84
,共5页
潘劲松%韩悦%陈丹鹏%徐磊%齐颖新%严志强
潘勁鬆%韓悅%陳丹鵬%徐磊%齊穎新%嚴誌彊
반경송%한열%진단붕%서뢰%제영신%엄지강
基质金属蛋白酶9%MAP激酶信号系统%周期性张应变%牙周膜细胞
基質金屬蛋白酶9%MAP激酶信號繫統%週期性張應變%牙週膜細胞
기질금속단백매9%MAP격매신호계통%주기성장응변%아주막세포
Matrix metalloproteinases-9%MAP kinase signaling system%Cyclic strain%Periodontal ligament cells
目的 探讨周期性张应变对人牙周膜细胞(human periodontal ligament cells,hPDLC)迁移的影响及其相关机制,为进一步了解机械力对hPDLC功能的影响提供资料.方法 应用FX-4000T细胞应变加载系统,对体外培养的hPDLC施加周期性张应变,牵张幅度分别为10%和20%,加载时间为6 h和24 h,频率均为0.1 Hz,以未加载的静态细胞作为对照组,应用划痕法观察hPDLC的迁移,蛋白质印迹法检测hPDLC的基质金属蛋白酶9(matrix metalloproteinases-9,MMP-9)的表达变化;用细胞外信号调节蛋白激酶(extracellular signal-regulated kinase,ERK)的特异性抑制剂PD98059预处理hPDLC后,在0.1 Hz、10%幅度条件下,加载6 h,检测细胞p-ERK1/2和MMP-9表达的变化.用细胞迁移划痕法观察加载10%和20%幅度张应变,观察24 h后PD98059和MMP家族的抑制剂强力霉素(doxycycline,DOX)对hPDLC迁移的影响.结果 细胞迁移划痕实验结果显示,牵张幅度和加载时间均可显著促进细胞迁移.与静态组相比,施加牵张幅度分别为10%和20%的张应变6 h后,hPDLC迁移变化不明显,但加载24 h后,10%和20%幅度的张应变均显著促进了hPDLC迁移(P<0.05),细胞迁移率分别为(45 ±8)%和(66±14)%,且20%比10%幅度牵张对细胞迁移的促进作用更显著(P<0.05).蛋白印迹法结果显示,周期性张应变促进了hPDLC的MMP-9表达.牵张幅度对细胞MMP-9的表达有显著影响(P<0.05),但加载时间对细胞MMP-9的表达无显著影响.ERK1/2的抑制剂PD98059不仅可以明显抑制张应变诱导的p-ERK1/2活化,而且可通过ERK信号通路明显抑制张应变诱导的细胞MMP-9表达.细胞迁移划痕实验还证实,加载幅度分别10%和20%的张应变24 h后,抑制剂PD98059和DOX均能有效抑制周期性张应变诱导的hPDLC迁移.结论 周期性张应变可通过激活hPDLC的ERK信号通路,促进细胞的MMP-9表达,从而诱导体外培养的hPDLC迁移.
目的 探討週期性張應變對人牙週膜細胞(human periodontal ligament cells,hPDLC)遷移的影響及其相關機製,為進一步瞭解機械力對hPDLC功能的影響提供資料.方法 應用FX-4000T細胞應變加載繫統,對體外培養的hPDLC施加週期性張應變,牽張幅度分彆為10%和20%,加載時間為6 h和24 h,頻率均為0.1 Hz,以未加載的靜態細胞作為對照組,應用劃痕法觀察hPDLC的遷移,蛋白質印跡法檢測hPDLC的基質金屬蛋白酶9(matrix metalloproteinases-9,MMP-9)的錶達變化;用細胞外信號調節蛋白激酶(extracellular signal-regulated kinase,ERK)的特異性抑製劑PD98059預處理hPDLC後,在0.1 Hz、10%幅度條件下,加載6 h,檢測細胞p-ERK1/2和MMP-9錶達的變化.用細胞遷移劃痕法觀察加載10%和20%幅度張應變,觀察24 h後PD98059和MMP傢族的抑製劑彊力黴素(doxycycline,DOX)對hPDLC遷移的影響.結果 細胞遷移劃痕實驗結果顯示,牽張幅度和加載時間均可顯著促進細胞遷移.與靜態組相比,施加牽張幅度分彆為10%和20%的張應變6 h後,hPDLC遷移變化不明顯,但加載24 h後,10%和20%幅度的張應變均顯著促進瞭hPDLC遷移(P<0.05),細胞遷移率分彆為(45 ±8)%和(66±14)%,且20%比10%幅度牽張對細胞遷移的促進作用更顯著(P<0.05).蛋白印跡法結果顯示,週期性張應變促進瞭hPDLC的MMP-9錶達.牽張幅度對細胞MMP-9的錶達有顯著影響(P<0.05),但加載時間對細胞MMP-9的錶達無顯著影響.ERK1/2的抑製劑PD98059不僅可以明顯抑製張應變誘導的p-ERK1/2活化,而且可通過ERK信號通路明顯抑製張應變誘導的細胞MMP-9錶達.細胞遷移劃痕實驗還證實,加載幅度分彆10%和20%的張應變24 h後,抑製劑PD98059和DOX均能有效抑製週期性張應變誘導的hPDLC遷移.結論 週期性張應變可通過激活hPDLC的ERK信號通路,促進細胞的MMP-9錶達,從而誘導體外培養的hPDLC遷移.
목적 탐토주기성장응변대인아주막세포(human periodontal ligament cells,hPDLC)천이적영향급기상관궤제,위진일보료해궤계력대hPDLC공능적영향제공자료.방법 응용FX-4000T세포응변가재계통,대체외배양적hPDLC시가주기성장응변,견장폭도분별위10%화20%,가재시간위6 h화24 h,빈솔균위0.1 Hz,이미가재적정태세포작위대조조,응용화흔법관찰hPDLC적천이,단백질인적법검측hPDLC적기질금속단백매9(matrix metalloproteinases-9,MMP-9)적표체변화;용세포외신호조절단백격매(extracellular signal-regulated kinase,ERK)적특이성억제제PD98059예처리hPDLC후,재0.1 Hz、10%폭도조건하,가재6 h,검측세포p-ERK1/2화MMP-9표체적변화.용세포천이화흔법관찰가재10%화20%폭도장응변,관찰24 h후PD98059화MMP가족적억제제강력매소(doxycycline,DOX)대hPDLC천이적영향.결과 세포천이화흔실험결과현시,견장폭도화가재시간균가현저촉진세포천이.여정태조상비,시가견장폭도분별위10%화20%적장응변6 h후,hPDLC천이변화불명현,단가재24 h후,10%화20%폭도적장응변균현저촉진료hPDLC천이(P<0.05),세포천이솔분별위(45 ±8)%화(66±14)%,차20%비10%폭도견장대세포천이적촉진작용경현저(P<0.05).단백인적법결과현시,주기성장응변촉진료hPDLC적MMP-9표체.견장폭도대세포MMP-9적표체유현저영향(P<0.05),단가재시간대세포MMP-9적표체무현저영향.ERK1/2적억제제PD98059불부가이명현억제장응변유도적p-ERK1/2활화,이차가통과ERK신호통로명현억제장응변유도적세포MMP-9표체.세포천이화흔실험환증실,가재폭도분별10%화20%적장응변24 h후,억제제PD98059화DOX균능유효억제주기성장응변유도적hPDLC천이.결론 주기성장응변가통과격활hPDLC적ERK신호통로,촉진세포적MMP-9표체,종이유도체외배양적hPDLC천이.
Objective To study the effect of cyclic strain on migration of human periodontal ligament cell(hPDLC) and underlying mechanism. Methods The cultured hPDLC were subjected to 10% or 20%-elongation magnitude cyclic strain at frequency of 0. 1 Hz by FX-4000T system for 6 or 24 hours-duration respectively, while the static group serves as control, hPDLC migration was assayed by wound healing method. The expressions of matrix metalloproteinases-9 (MMP-9) and p-ERK1/2 in hPDLC without or with cyclic srain were analyzed by Western blotting. To investigate the effect of ERK signaling pathway and MMP-9 on migration of hPDLC, the cells were incubated with PD98059, a specific extracellular signal-regulated kinase (ERK) kinase inhibitor, or doxycycline, a MMP inhibitor. Then the expressions of p-ERK1/2 and MMP-9 and hPDLC migration were analysized. Results In wound healing tests, the migration of hPDLC exposed to 10% or 20%-cyclic strain at 0. 1 Hz-frequency for 6 hours was not apparent but became significantly different for 24 hours (P <0. 05) compared to control. Furthermore, the 20%-elongation magnitude of cyclic strain had more remarkable effect on migration of hPDLC than 10%-elongation magnitude at 24 hours-duration(P <0. 05). Cyclic strain obviously increased the expression of MMP-9w in hPDLC(P <0. 05). PD98059 could repress not only the activation of p-ERK1/2 but also the expression of MMP-9 induced by eyclic strain in hPDLC. The migration of hPDLC enhanced by cyclic strain was repressed by DOX or PD98059 in wound healing tests. Conclusions Cyclic strain promotes the migration of hPDLC through activating ERK signaling pathway and inducing the expression of MMP-9.
