大鼠骨髓间充质干细胞和颅骨成骨细胞在张应力下细胞骨架的改变
The Changes of Cytoskeleton F-actin in Rat Bone Marrow Mesenchymal Stem Cells and Calvarial Osteoblasts under Me-
chanical Strain
作者：戚孟春,胡静,邹淑娟,韩力赤,罗恩
Author：QIMeng-chun, HUJing, ZOUShu-juan, HANLi-chi, LUO En
收稿日期：2005-04-25 年卷（期）页码：2005,23(02):110-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：间充质干细胞,成骨细胞,机械张应力,激光共聚焦显微镜,
Key words：mesenchymal stem cells,osteoblasts,mechanical stretch,laser scanning confocal microscope,
基金项目：
国家自然科学基金资助项目(30371553);高等学校博士学科点专项科研基金资助项目(20020610062)
中文摘要
目的 探讨大鼠骨髓间充质干细胞(MSCs)和颅骨成骨细胞(OB)在机械张应力刺激下的反应和细胞骨架蛋白F-actin的变化。方法 体外分离培养大鼠骨髓MSCs和颅骨OB,应用四点弯曲加力系统对细胞施加2 000με 的机械张应力,加力时间分别为0 h、2 h、6 h、12 h。采用激光共聚焦显微镜观察特异性荧光染料标记的微丝蛋白 F-actin和细胞核,并用图像分析软件对细胞形态学参数进行测量。结果 MSCs和OB荧光强度减弱,F-actin纤维束稀疏,排列紊乱;胞核轮廓模糊,部分细胞可观察到凋亡小体;MSCs体积明显变小,OB体积变化不明显。定量分析表明, MSCs的细胞总面积、总荧光密度、绿色荧光(F-actin)密度均显著降低(P
英文摘要
Objective To explore the response of rat bone marrowmesenchymal stem cells (MSCs) and calvarial osteoblasts to mechanical strain and the consequent changes of cytoskeleton F-actin.Methods Bone marrow MSCs and calvarial osteoblasts were isolated from SD rats and culturedin vitro. Mechanical stretchwas performed on passage 3 cells at 2 000μεfor 0, 2, 6 and 12 hours using four-point bending system. The response of cells and the distribution of F-actin were observed using fluorescent staining under laser scanning confocal microscope and the morphological parameters were quantified using image analysis software Laserpix.Results Under mechanical stretch, the fluorescent staining decreased obviously at bothMSCs and osteoblasts, and F- actin filaments were rearranged and became tenuous, thinner, and abnormally distributed. The outline of nucleus became unclear and apoptotic changes were observed at some of both cells. Cellular size decreased more significantly inMSCs than in osteoblasts. Quantity analysis showed that total area of cells, total fluorescent density and green fluorescent density (F-actin) were all signifi- cantly decreased in MSCs(P
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