肌损伤浸液对骨髓间充质干细胞增殖及其成肌诱导作用(英文)  

Inductive effect of muscular traumatic fluid on the proliferation and myogenesis of bone marrow mesenchymal stem cells

作　　者：王劲[1] 罗成基 冉新泽[2] 许利民[1] 冯江[1] 

机构地区：[1]解放军第三军医大学大坪医院野战外科研究所血液科,重庆市400042 [2]解放军第三军医大学预防医学院复合伤研究所创伤,烧伤与复合伤国家重点实验室,重庆市400038

出　　处：《中国临床康复》2005年第18期270-272,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家规划"九七三"基金资助项目(G1999054205);全军"十五"医药卫生科研基金(青年基金)(01Q110)~~

摘　　要：背景:骨髓间充质干细胞具有可诱导分化为成骨细胞及成软骨细胞,利用DNA甲基化抑制剂5-氮杂胞苷可诱导间充质干细胞表达生肌调控因子中的Myf5及生肌素,可以使间充质干细胞定向分化为成肌细胞。目的:通过筛选蛋白含量最高时相的肌损伤浸液作用于骨髓间充质干细胞,探讨肌损伤浸液对间充质干细胞增殖及成肌诱导作用。设计:标准对照实验。单位:解放军第三军医大学预防医学院复合伤研究所,创伤、烧伤与复合伤国家重点实验室。材料:Balb/C纯系小鼠18只造成肌肉组织的创伤模型,用于肌损伤浸液的提取,同时以5-氮杂胞苷诱导的间充质干细胞成肌组进行对照。方法:实验于2001-04/2003-09在第三军医大学预防医学院全军复合伤研究所,创伤、烧伤及复合伤国家重点实验室进行。Bradford比色法检测不同时相肌浸液蛋白含量,筛选蛋白含量最高时相的肌损伤浸液作用于骨髓间充质干细胞,于0,3,6,9,12,15d采用四甲基偶氮唑盐法检测伤浸液对间充质干细胞增殖的影响,与未加浸液的间充质干细胞作对照。采用反转录聚合酶链反应法检测其作用后6d生肌素的表达,并与用5-氮杂胞苷诱导的间充质干细胞成肌分化进行对照。主要观察指标:Balb/c小鼠①肌损伤浸液蛋白含量测定。②肌损伤浸液对间充质干细胞增殖的影响。③以生肌素的表达观察肌损伤浸液对间充质干细胞的成肌诱导作用。结果:①肌损伤浸液蛋白含量分析:以损伤后12h为最高。②肌损伤浸液对间充质干细胞增殖的影响:伤浸液作用组间充质干细胞的增殖活力在各时相均明显高于对照组。③肌损伤浸液对间充质干细胞的成肌诱导作用:伤浸液作用组无生肌素的表达,5-氮杂胞苷作用组有较强的生肌素表达。结论:肌损伤浸液可促进间充质干细胞的增殖,但对间充质干细胞无成肌诱导分化作用。BACKGROUND: Bone marrow mesenchymal stem cells(BMMSCs) can be induced differ entiating into osteoblasts and chondroblasts, DNA methylate depressant 5- aza cytidine can induce BMMSCs expressing myogenic regulating factors: Myf5 and myop oietin, which involving in the differentiation of BMMSCs into myoblasts. OBJECTIVE: Muscular traumatic fluid containing the highest protein content wa s screened out and co- cultured with BMMSCs, in order to explore the inductive effect on the proliferation and myogeneis of BMMSCs. DESIGN: Standardized comparative study. SETTING: At State Key Laboratory of Trauma, Burns and Combined Injury, Insti tute of Combined Injury,Medical College of Preventive Medicine, Third Military M edical University of Chinese PLA. MATERIALS: Muscular traumatic model was established on 18 Balb/C pure rats f or the extraction of muscular traumatic fluid, the inductive effect of the flui d on BMMSC was then compared with 5- azacytidine. METHODS: This study was carried out at State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, College of Preventive Medici ne, Third Military Medical University of Chinese PLA from April 2001 and Septemb er 2003.Bradford colorimetric was used to detect the protein content in the musc ular traumatic fluid,and the fluid with the highest protein was used to co- cul ture with BMMSC, whose effect on the proliferation of BMMSC was measured with M TT methods at day 0, 3, 6, 9, 12, 15. RT- PCR technique was used to detect the expression of myopoietin at day 6, with its myogenic effect compared with that of 5- azacytidine. MAIN OUTCOME MEASURES: ① The protein content in the muscular traumatic flui d;② The influence of the fluid on the proliferation of BMMSC; ③ The expression of myopoietin and the myogenic inducement of traumatic fluid on BMMSC. RESULTS: ① The protein content in the traumatic fluid: It reached to the pe ak at post- traumatic 12 hours.② The influence of the fluid on the proliferati on of BMMSC: the proloferative activity of

关 键 词：骨髓细胞 干细胞 细胞培养 肌/损伤 组织 培养基 

分 类 号：E329.2[军事—军事理论]