中文摘要：

研究目的：探讨不同浓度红景天苷对成肌细胞株C2C12体外分化的影响及可能机制,有望为成肌细胞增殖分化寻求更有效的外源调控物,以促进成肌细胞的移植和临床应用,同时为进一步开展红景天苷在体内骨骼肌损伤中的作用及机制研究奠定基础。研究方法：研究1：红景天苷对成肌细胞体外分化的影响：1）在C2C12中加入不同浓度红景天苷处理,在诱导成肌分化过程中,应用相差显微镜观察不同浓度红景天苷对C2C12分化过程中汇聚、融合和形成多核肌管的影响;2）应用免疫荧光组化方法,分析不同浓度红景天苷对C2C12成肌分化融合率的影响;3）应用real-time PCR和Western Blotting方法,检测红景天苷对C2C12成肌分化特异基因和蛋白表达的影响;研究2：红景天苷影响成肌细胞体外分化的机制初探：应用Western Blotting方法,检测红景天苷处理后C2C12成肌分化中TGF-β/Smad信号通路的变化。结果：50μg红景天苷处理：研究1：1）光镜观察结果从形态学上显示,红景天苷处理显著抑制C2C12细胞的融合和多核肌管的形成;2）免疫荧光检测结果表明Myosin-neonatal（E-MHC）阳性面积和myogenin阳性核均显著减少（P〈0.05）,显著抑制C2C12细胞分化过程中肌管形成,促进细胞增殖;3）real-time PCR和Western blotting检测结果表明,成肌分化特异因子MyoD和myogenin基因和蛋白表达水平均显著下降（P〈0.05）,显著抑制C2C12细胞分化过程中细胞的融合和肌管的形成;研究2：Western blotting检测结果表明,红景天苷显著抑制C2C12细胞分化效应,伴有Smad2/Smad3磷酸化激活增加,尤其是Smad3激活水平显著增加（P〈0.01）;30μg红景天苷处理作用不显著（P〉0.05）。结论：1）红景天苷能显著抑制骨骼肌成肌细胞体外成肌分化,并促进骨骼肌成肌细胞激活增殖,促进卫星细胞库储备增加。2）红景天苷对骨骼肌成肌细胞体外分化增殖的?

英文摘要：

Objective： This study is aim to explore the effect of different concentrations of sali- droside on myoblast differentiation in vitro and its possible mechanism, which can help to find a more effective exogenous regulating substance for myoblast differentiation to promote the myo blast transplantation, and set the foundation for further study of the effect of salidroside on skeletal muscle in vivo. Methods：Part one：The effect of different concentrations of salidroside on myoblast differentiation in vitro： 1）observe the morphology of differential myoblast C2C12 which were treated with 0,30,50 μg/ml salidroside to induce differentiation by Phase micro- graphs; 2）Detect the cell fusion index affected by salidroside using immunofluorescence; 3）Detect the expressions of myogenic differentiation-specific mRNAs and proteins affected by sali- droside using Western Blotting and real time PCR. Part two：The possible mechanism of above study ：Detect TGF-β/Smad3 pathway protein expression in differential C2C12 cell treated with 0,30,50μg/ml Sal using Western Blotting. Results ： Treated with 50 /,g/ml salidroside ： Part one： 1）Salidroside inhibit C2C12 fusion and myotube form in morphological by Phase mi- crographs 2）Myosin-neonatal Positive area and myogenin Positive Nuclei decreased significantly by irnmunofluoreseence （ P 〈 0. 05 ）, which inhibited myotube form notably ; 3 ） The expres- sions of MyoD and myogenin in mRNA and protein both decreased notably by real-time PCR and Western blotting（P〈0.05）, which inhibited cell fusion and myotube form during C2C12 differentiation. Part two： Salidroside inhibit C2C12 differentiation associated with increases in Smad2 / Smad3 phosphorylation, especially p-Smad3 （ P 〈 0. 01 ）. The results treated with 30 μg/ml salidroside is insignificant （P〈0.05）. Conclusion： 1）Salidroside inhibit myogenic dif-ferentiation of C2C12 cell significantly, and promote cell proliferation, to add satellite cells band reserve. 2 ）Salidroside