流体静压力调控下Rac1信号通路在大鼠BMSCs成软骨响应中的作用.pdfVIP

流体静压力调控下 Rac1信号通路在大鼠 BMSCs成软骨响应 中的作用 易飞舟;赵萤;赵寅华;张旻 【摘要】目的：：观察流体静压力作用下 Rac1 信号通路对大鼠骨髓间充质干细 胞( BMSCs)增殖、细胞骨架改建及成软骨向分化的影响。方法：体外分离培养大 鼠 BMSCs,并随机分为对照组、Rac1 激动剂组、Rac1 拮抗剂组、压力组、Rac1 激动剂 +压力组、Rac1 拮抗剂 +压力组；其中各压力组均置于细胞加载装置中施 加 90 kPa 流体静压力,每天 1 h,其他各组常规条件下进行培养。分别对各组细胞处 理后,采用流式细胞仪检测细胞周期、共聚焦显微镜观察细胞骨架、Real-time PCR 检测成软骨标志基因 ( Sox9、Aggrecan、Col-ⅡmRNA)的表达水平。结果： 流体静压力下 BMSCs 中 DNA 合成期的细胞比例及细胞增殖指数显著高于对照组 (P≤0.05),且该过程依赖 Rac1 活性；而 Rac1 在流体静压力导致的细胞骨架光密度 值升高(P≤0.05)的过程中呈负调控作用；流体静压力可促 BMSCs 中成软骨标志基 因的表达量显著升高(P≤0.05),该过程依赖 Rac1 活性。结论：Rac1 在流体静压力 导致的细胞周期启动、骨架装配及成软骨分化的过程中均具有调控作 用。%AIM:To study the role of Racl signal pathway in chondrogenic transduction of bone marrow mes-enchymal stem cells ( BMSCs) under hydrostatic pressure. METHODS: BMSCs were isolated and cultured in vitro. The cells were then randomly divided into control group, Rac1agonist group, Rac1antagonist group, hydrostatic pres-sure group, hydrostatic pressure plus Aac1 agonist group and pressure plus Aac1 antagonist group. Cell cycle was ex-amined by flow-cytometry and cytoskeleton was observed by confocal microscopy. mRNA expression of Sox9, Aggrecan and Col- Ⅱ were detected by real-time PCR. RESULTS: Under hydrostatic pressure, S-phase fraction and the cell proliferation index of BMSCs were significantly higher than those in the control group (P≤0. 05) in a Rac1- dependent way. However, activation of the Rac1 pathway blocked F-actin cytoskeleton assembly which induced by hydrostatic pressure. The significant increase of chondrogenic marker mRNA expression in chondrogenic differentiation induced by hydrostatic pressure (P≤0. 05) was dependent on the activity of Rac1. CONCLUSION:Rac1 plays an important role in the start of the BMSCs cell cycle, cytoskeleton assembly and chondrogenic differentiation under mechanical pressure. 【期刊名称】《牙体牙髓牙周病学杂志》 【年(卷),期】2015(000)006 【总页数】7 页(P335-341) 【关键词】流体静压力;骨髓间充质干细胞;Rac1;细胞周期;细胞骨架;成软骨分化 【作者】易飞舟;赵萤;赵寅华;张旻 【作者