牙体牙髓牙周病学杂志
牙體牙髓牙週病學雜誌
아체아수아주병학잡지
CHINESE JOURNAL OF CONSERVATIVE DENTISTRY
2015年
6期
335-341
,共7页
易飞舟%赵萤%赵寅华%张旻
易飛舟%趙螢%趙寅華%張旻
역비주%조형%조인화%장민
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
流體靜壓力%骨髓間充質榦細胞%Rac1%細胞週期%細胞骨架%成軟骨分化
류체정압력%골수간충질간세포%Rac1%세포주기%세포골가%성연골분화
hydrostatic pressure%bone marrow mesenchymal stem cells ( BMSCs)%Ras-related C3 botulinum toxin substrate 1 (Rac1)%proliferation%cell cytoskeleton%chondrogenic differentiation [Chinese Journal of Conservative Dentistry,2015,25(6):335
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。
目的：：觀察流體靜壓力作用下Rac1信號通路對大鼠骨髓間充質榦細胞( BMSCs)增殖、細胞骨架改建及成軟骨嚮分化的影響。方法：體外分離培養大鼠BMSCs,併隨機分為對照組、Rac1激動劑組、Rac1拮抗劑組、壓力組、Rac1激動劑+壓力組、Rac1拮抗劑+壓力組；其中各壓力組均置于細胞加載裝置中施加90 kPa流體靜壓力,每天1 h,其他各組常規條件下進行培養。分彆對各組細胞處理後,採用流式細胞儀檢測細胞週期、共聚焦顯微鏡觀察細胞骨架、Real-time PCR檢測成軟骨標誌基因( Sox9、Aggrecan、Col-ⅡmRNA)的錶達水平。結果：流體靜壓力下BMSCs中DNA閤成期的細胞比例及細胞增殖指數顯著高于對照組(P≤0.05),且該過程依賴Rac1活性；而Rac1在流體靜壓力導緻的細胞骨架光密度值升高(P≤0.05)的過程中呈負調控作用；流體靜壓力可促BMSCs中成軟骨標誌基因的錶達量顯著升高(P≤0.05),該過程依賴Rac1活性。結論：Rac1在流體靜壓力導緻的細胞週期啟動、骨架裝配及成軟骨分化的過程中均具有調控作用。
목적：：관찰류체정압력작용하Rac1신호통로대대서골수간충질간세포( BMSCs)증식、세포골가개건급성연골향분화적영향。방법：체외분리배양대서BMSCs,병수궤분위대조조、Rac1격동제조、Rac1길항제조、압력조、Rac1격동제+압력조、Rac1길항제+압력조；기중각압력조균치우세포가재장치중시가90 kPa류체정압력,매천1 h,기타각조상규조건하진행배양。분별대각조세포처리후,채용류식세포의검측세포주기、공취초현미경관찰세포골가、Real-time PCR검측성연골표지기인( Sox9、Aggrecan、Col-ⅡmRNA)적표체수평。결과：류체정압력하BMSCs중DNA합성기적세포비례급세포증식지수현저고우대조조(P≤0.05),차해과정의뢰Rac1활성；이Rac1재류체정압력도치적세포골가광밀도치승고(P≤0.05)적과정중정부조공작용；류체정압력가촉BMSCs중성연골표지기인적표체량현저승고(P≤0.05),해과정의뢰Rac1활성。결론：Rac1재류체정압력도치적세포주기계동、골가장배급성연골분화적과정중균구유조공작용。
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.
