肌损伤浸液对骨髓间充质干细胞增殖及其成肌诱导作用

背景骨髓问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5一氮杂胞昔可诱导问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问充质干细胞,探讨肌损伤浸液对问充质于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5一氮杂胞昔诱导的问充质干细胞成肌组进行对照.方法实验于2001一04/200309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采用四甲基偶氮哩盐法检测伤浸液对问充质于细胞增殖的影响,与未加浸液的问充质干细胞作对照.采用反转录聚合酶链反应法检测其作用后66生肌素的表达,并与用5一氮杂胞音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肌损伤浸液对问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问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,5一氮杂胞耷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于细胞的增殖,但对问充质干细胞无成肌诱导分化作用.

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

BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) can be induced differentiating into osteoblasts and chondroblasts, DNA methylate depressant 5-azacytidine can induce BMMSCs expressing myogenic regulating factors: Myf5 and myopoietin, which involving in the differentiation of BMMSCs into myoblasts.OBJECTIVE: Muscular traumatic fluid containing the highest protein content was 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, Bums and Combined Injury,Institute of Combined Injury, Medical College of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: Muscular traumatic model was established on 18 Balb/C pure rats for the extraction of muscular traumatic fluid, the inductive effect of the fluid 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 Medicine, Third Military Medical University of Chinese PLA from April 2001 and September 2003. Bradford colorimetric was used to detect the protein content in the muscular traumatic fluid, and the fluid with the highest protein was used to co-culture with BMMSC, whose effect on the proliferation of BMMSC was measured with MTT 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.fluid on BMMSC.eration of BMMSC: the proloferative activity of BMMSC in traumatic fluid genic effect of traumatic fluid on BMMSC: myopoietin could not be found expressed in traumatic fluid group, but strongly expressed in 5-azacytidine group.CONCLUSION: Muscular traumatic fluid can promote the proliferation of BMMSC, but has no myogenic effect.