中华骨科杂志
中華骨科雜誌
중화골과잡지
CHINESE JOURNAL OF ORTHOPAEDICS
2012年
5期
494-499
,共6页
李稚君%马信龙%付鑫%马剑雄%张华峰%孙晓雷%李鸿雁%赵秀宝
李稚君%馬信龍%付鑫%馬劍雄%張華峰%孫曉雷%李鴻雁%趙秀寶
리치군%마신룡%부흠%마검웅%장화봉%손효뢰%리홍안%조수보
应力,物理%成骨细胞%蛋白激酶C
應力,物理%成骨細胞%蛋白激酶C
응력,물리%성골세포%단백격매C
Stress,mechanical%Osteoblasts%Protein kinase C
目的 探讨蛋白激酶Cξ(protein kinases Cξ,PKCξ)蛋白通路参与响应调控周期性牵张应力促进成骨细胞增殖的相关机制.方法 无菌环境下,组织分离法培养SD大鼠成骨细胞,应用BioDynamic细胞应力加载系统对第三代大鼠成骨细胞进行单轴周期性牵张应力加载,形变量为2%,加载频率为0.25 Hz,单轴向周期性牵张应力180 min(即实验组);应用同样的方法加载以PKCξ特异性经典抑制剂Myristoylate预处理的成骨细胞(即经典抑制剂组);使用同样的培养加载方式形变量为0%单轴向周期性牵张应力180 min(即静态对照组).应用流式细胞术检测大鼠成骨细胞的细胞周期表达,并使用免疫印迹法(Western blotting)、免疫荧光法分别检测实验组、经典抑制剂组和静态对照组大鼠成骨细胞内转导蛋白通路PKCξ与磷酸化PKCξ(p-PKCξ)的表达.结果 与静态对照组、经典抑制剂组相比,单轴向周期性牵张应力可以使大鼠成骨细胞S期显著前移,并显著上调大鼠成骨细胞转导蛋白通路PKCξ与p-PKCξ的表达;同时,抑制剂可以明显抑制p-PKCξ活化,从而抑制PKCξ的表达.结论 成骨细胞PKCξ蛋白通路参与感知周期性牵张应力刺激,并调控促进大鼠成骨细胞增殖,是力学信号传导通路中重要的环节.
目的 探討蛋白激酶Cξ(protein kinases Cξ,PKCξ)蛋白通路參與響應調控週期性牽張應力促進成骨細胞增殖的相關機製.方法 無菌環境下,組織分離法培養SD大鼠成骨細胞,應用BioDynamic細胞應力加載繫統對第三代大鼠成骨細胞進行單軸週期性牽張應力加載,形變量為2%,加載頻率為0.25 Hz,單軸嚮週期性牽張應力180 min(即實驗組);應用同樣的方法加載以PKCξ特異性經典抑製劑Myristoylate預處理的成骨細胞(即經典抑製劑組);使用同樣的培養加載方式形變量為0%單軸嚮週期性牽張應力180 min(即靜態對照組).應用流式細胞術檢測大鼠成骨細胞的細胞週期錶達,併使用免疫印跡法(Western blotting)、免疫熒光法分彆檢測實驗組、經典抑製劑組和靜態對照組大鼠成骨細胞內轉導蛋白通路PKCξ與燐痠化PKCξ(p-PKCξ)的錶達.結果 與靜態對照組、經典抑製劑組相比,單軸嚮週期性牽張應力可以使大鼠成骨細胞S期顯著前移,併顯著上調大鼠成骨細胞轉導蛋白通路PKCξ與p-PKCξ的錶達;同時,抑製劑可以明顯抑製p-PKCξ活化,從而抑製PKCξ的錶達.結論 成骨細胞PKCξ蛋白通路參與感知週期性牽張應力刺激,併調控促進大鼠成骨細胞增殖,是力學信號傳導通路中重要的環節.
목적 탐토단백격매Cξ(protein kinases Cξ,PKCξ)단백통로삼여향응조공주기성견장응력촉진성골세포증식적상관궤제.방법 무균배경하,조직분리법배양SD대서성골세포,응용BioDynamic세포응력가재계통대제삼대대서성골세포진행단축주기성견장응력가재,형변량위2%,가재빈솔위0.25 Hz,단축향주기성견장응력180 min(즉실험조);응용동양적방법가재이PKCξ특이성경전억제제Myristoylate예처리적성골세포(즉경전억제제조);사용동양적배양가재방식형변량위0%단축향주기성견장응력180 min(즉정태대조조).응용류식세포술검측대서성골세포적세포주기표체,병사용면역인적법(Western blotting)、면역형광법분별검측실험조、경전억제제조화정태대조조대서성골세포내전도단백통로PKCξ여린산화PKCξ(p-PKCξ)적표체.결과 여정태대조조、경전억제제조상비,단축향주기성견장응력가이사대서성골세포S기현저전이,병현저상조대서성골세포전도단백통로PKCξ여p-PKCξ적표체;동시,억제제가이명현억제p-PKCξ활화,종이억제PKCξ적표체.결론 성골세포PKCξ단백통로삼여감지주기성견장응력자격,병조공촉진대서성골세포증식,시역학신호전도통로중중요적배절.
Objective To investigate the proliferation of rat's osteoblasts promoted by mechanical strain via the protein kinases C ξ (PKCξ) protein path-way.Methods Osteoblasts were retrieved from SD rats' skulls in the sterilized environment.BioDynamic testing instrument was used to exert 2% and 0% mechanical strain on rats' osteoblasts for 180 min on the each group (2% mechanical strain was experimental group and 0% mechanical strain was control group); and the same method was applied on the rats' osteoblasts which pretreated with the classical PKC ξ inhibitor with 2% mechanical strain (inhibitor group).The cell cycles of rats' osteoblasts were measured by flow cytometry; and Western blotting and immunofluorescence were used to detect expression of protein PKC ξ and phosphorylationed PKC ξ in the rats' osteoblasts.Results The mechanical strain obviously increased the ratio of S period in the cell cycles.Compared to the control group and inhibitor group,a significant increase of the expression of protein PKC ξ and phosphorylationed PKC ξ in the osteoblasts was detected in the mechanical strain experimental group.ConclusionPKCξ can respond to the stimulus of the mechanical strain,which promotes the proliferation of osteoblasts through PKCξ pathway.And it plays an important role in the process of signal conduction.