应用单分子荧光成像技术研究人参抗运动性疲劳分子机制

Application of Cytoplasmic Calcium Fluorescence Imaging Techniques to Study the Molecular Mechanisms of the Elimination of Exercise-induced Fatigue by Chinese Herb Ginseng

采用单分子荧光成像技术,通过建立运动性疲劳的细胞模型对中药人参抗运动性疲劳的分子机制进行研究。取新生大鼠骨骼肌细胞,将其随机分成人参组和对照组。在两组细胞的培养基中分别加入2 mg/mL人参提取物和等量的D-hanks液进行培养。两组细胞分化成肌管后,将用钙离子染料处理过的细胞置于共聚焦显微镜上检测其荧光强度。待荧光强度稳定时,分别向两组细胞加1μmol/L地塞米松溶液,记录并比较两组细胞用地塞米松刺激前后荧光强度的改变幅度。结果发现,两组细胞胞浆内钙离子的荧光强度均明显增强(P<0.05),但人参提取物组细胞胞浆内钙离子浓度的增加幅度明显低于对照组(P<0.05)。由此可见,人参提取物对地塞米松刺激引起的疲劳骨骼肌细胞具有保护作用,人参消除运动性疲劳的机制可能与其调节骨骼肌细胞胞浆内钙离子浓度有关。

Through seting up the cell models of exercise-induced fatigue, we study the molecular mechanisms of exercise-induced fatigue eliminated by Chinese herb ginseng extract with single-molecule fluorescence imaging techniques. The skeletal muscle cells from the newborn rats were randomly divided into the ginseng group and the control group. The cells from the two groups were cultured in the medium respectively added 2 mg/mL ginseng extract and 2 mg/mL D- hanks. After differentiating into myotubes, the two groups of cells treated with a fluorescent probe Fluo-3 AM were put on the confocal microscope and the fluorescence intensity of cells were detected. Once the fluorescence intensity was sta-ble, 1 μmol/L dexamethasone were added into the cells from the two groups, then the variation amplitude of fluorescence intensity of the two groups cells were recorded and compared. The results showed that the fluorescence intensity of the two groups cells were markedly increased （ P 〈 0.05 ） , however, the increasing amplitude of fluorescence intensity of the ginseng extract group were significantly lower than that of the control group. It was concluded that the ginseng have a protective effect on the skeletal muscle cells stimulated by dexamethasone. The molecular mechanisms of exercise-induced fatigue eliminated by Chinese medicine ginseng may have a relation to the effect of regulating calcium concentration of skeletal muscle cells.