中华口腔医学研究杂志（电子版）
中華口腔醫學研究雜誌（電子版）
중화구강의학연구잡지（전자판）
CHINESE JOURNAL OF STOMATOLOGICAL RESEARCH(ELECTRONIC VERSION)
2008年
4期
352-358
,共7页
机械应力%细胞外信号调节激酶%酪氨酸蛋白激酶%细胞松弛素
機械應力%細胞外信號調節激酶%酪氨痠蛋白激酶%細胞鬆弛素
궤계응력%세포외신호조절격매%락안산단백격매%세포송이소
Osteoblast%Extracellular signal regulated kinase%Tyrosine protein kinase%Cytochalasin
目的 观察机械应力下酪氨酸蛋白激酶(TPK)抑制剂和细胞松弛素D对体外培养的成骨样细胞细胞外信号调节激酶(ERK)1/2早期表达变化的影响.方法 采用四点弯曲细胞力学加载装置系统,对人成骨样细胞进行加力:频率0.5 Hz,加力板变形量4000 μstrain牵张应力作用成骨样细胞(MG-63)5 min时,使细胞发生单轴牵张和压缩应变.运用Western免疫印迹检测不同方向应力应变下TPK抑制剂和细胞松弛素D对ERK1/2表达变化的影响.结果 TPK抑制剂预处理后,牵张应力激活ERK的作用被明显阻断,与单纯张力组相比差异有统计学意义(P ＜ 0.01);而压缩应力的诱导作用未受影响(P ＞ 0.05);低剂量的细胞松弛素D处理后,压应力的诱导作用被明显阻断,ERK的磷酸化水平甚至低于基态,与单纯压力组相比差异有统计学意义(P ＜ 0.01),而牵张应力对ERK的激活仅受到一定程度的影响,与单纯张应力组相比差异有统计学意义(P ＜ 0.05).结论 力学信号可同时激活细胞内的细胞骨架、TPK转导通道,但各自主要激活的部分存在差别,无论是牵张还是压缩,成骨细胞对力学信号的感知和转导都与微丝细胞骨架密切相关,TPK的活化可能是牵张应力引起黏附斑复合物形成后的主要后续反应,压缩应力的主要后续反应与之关系不大.
目的 觀察機械應力下酪氨痠蛋白激酶(TPK)抑製劑和細胞鬆弛素D對體外培養的成骨樣細胞細胞外信號調節激酶(ERK)1/2早期錶達變化的影響.方法 採用四點彎麯細胞力學加載裝置繫統,對人成骨樣細胞進行加力:頻率0.5 Hz,加力闆變形量4000 μstrain牽張應力作用成骨樣細胞(MG-63)5 min時,使細胞髮生單軸牽張和壓縮應變.運用Western免疫印跡檢測不同方嚮應力應變下TPK抑製劑和細胞鬆弛素D對ERK1/2錶達變化的影響.結果 TPK抑製劑預處理後,牽張應力激活ERK的作用被明顯阻斷,與單純張力組相比差異有統計學意義(P ＜ 0.01);而壓縮應力的誘導作用未受影響(P ＞ 0.05);低劑量的細胞鬆弛素D處理後,壓應力的誘導作用被明顯阻斷,ERK的燐痠化水平甚至低于基態,與單純壓力組相比差異有統計學意義(P ＜ 0.01),而牽張應力對ERK的激活僅受到一定程度的影響,與單純張應力組相比差異有統計學意義(P ＜ 0.05).結論 力學信號可同時激活細胞內的細胞骨架、TPK轉導通道,但各自主要激活的部分存在差彆,無論是牽張還是壓縮,成骨細胞對力學信號的感知和轉導都與微絲細胞骨架密切相關,TPK的活化可能是牽張應力引起黏附斑複閤物形成後的主要後續反應,壓縮應力的主要後續反應與之關繫不大.
목적 관찰궤계응력하락안산단백격매(TPK)억제제화세포송이소D대체외배양적성골양세포세포외신호조절격매(ERK)1/2조기표체변화적영향.방법 채용사점만곡세포역학가재장치계통,대인성골양세포진행가력:빈솔0.5 Hz,가력판변형량4000 μstrain견장응력작용성골양세포(MG-63)5 min시,사세포발생단축견장화압축응변.운용Western면역인적검측불동방향응력응변하TPK억제제화세포송이소D대ERK1/2표체변화적영향.결과 TPK억제제예처리후,견장응력격활ERK적작용피명현조단,여단순장력조상비차이유통계학의의(P ＜ 0.01);이압축응력적유도작용미수영향(P ＞ 0.05);저제량적세포송이소D처리후,압응력적유도작용피명현조단,ERK적린산화수평심지저우기태,여단순압력조상비차이유통계학의의(P ＜ 0.01),이견장응력대ERK적격활부수도일정정도적영향,여단순장응력조상비차이유통계학의의(P ＜ 0.05).결론 역학신호가동시격활세포내적세포골가、TPK전도통도,단각자주요격활적부분존재차별,무론시견장환시압축,성골세포대역학신호적감지화전도도여미사세포골가밀절상관,TPK적활화가능시견장응력인기점부반복합물형성후적주요후속반응,압축응력적주요후속반응여지관계불대.
Objective To study the initial responses and the effect of mechanical strain with tyrosine protein kinase TPK and cytochalasin D on the expression of ERK1/2 in osteoblast-like cells in vitro. Methods Osteoblast-like cells were cultured with tyrosine protein kinase TPK and cytochalasin D and then subjected to mechanical strain by self-made four-point bending system at a0.5Hz frequency for 5 min, In each time-phase, the cells were loaded with tension and compression stress at 4000 μstrain respectively. The phosphorylation levels of ERK1/2 were measured by Western Blotting. Changes in the experiments were expressed as ratio to the controls(P ＜ 0.05). Results With tyrosine protein kinase TPK blocking agent 4000 μstrain tensile stimulus could induce the expression of ERK in a few minutes, while compressive stimulus could not. On the contrary, the effect with cytochalasin D of compressive tension strain stimulus was more significant. Conclusions The reception and transduction of osteoblasts to the mechanical signal of tension or compressive strain are relative to the microfilament cytoskeleton, while the activation of tyrosine protein kinases is mainly connected to response to the tension strain.
