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目的探讨大鼠骨骼肌挫伤后损伤区外周血来源的纤维细胞数量变化规律。方法制作大鼠骨骼肌挫伤模型,应用CD45、procollagenⅠ双重免疫荧光染色法检测大鼠骨骼肌挫伤后各时间点外周血来源的纤维细胞的变化。结果对照组大鼠骨骼肌中未见CD45、procollagenⅠ阳性的纤维细胞分布;损伤组大鼠骨骼肌挫伤后6 h～1 d,仅见少数CD45阳性的单个核细胞;伤后3 d,开始有少量CD45、procollagenⅠ阳性的纤维细胞聚集于损伤区;伤后5d,阳性的纤维细胞数量明显增加;伤后7d达高峰,此后逐渐减少,同时还发现相对于3 d组,纤维细胞的体积随着时间延长逐渐变大;伤后14 d,纤维细胞胞质中的CD45和pro-collagenⅠ的表达强度都相应减弱。结论大鼠骨骼肌挫伤后,挫伤区外周血来源的纤维细胞的数量变化具有一定的时间规律性,可望作为推断骨骼肌挫伤时间的指标之一。
Objective To investigate the changes of peripheral blood-derived fibroblasts in injured area of skeletal muscle of rats. Methods Skeletal muscle contusion model was made in rats. CD45 and procollagen Ⅰ double immunofluorescence staining were used to detect the changes of peripheral blood-derived fibroblasts at each time point after contusion of skeletal muscle in rats. Results There were no CD45 and procollagen Ⅰ-positive fibroblasts in the skeletal muscle of rats in the control group. Only a few CD45 positive mononuclear cells were found in the skeletal muscle of rats in injury group from 6 h to 1 d after contusion; CD45, procollagen I-positive fibroblasts gathered in the injured area; 5d after injury, the number of positive fibroblasts increased significantly; reached a peak on the 7d after injury, and then gradually decreased, and also found that relative to the 3d group, the volume of fibroblasts over time After 14 days of injury, the expression of CD45 and pro-collagen I in the cytoplasm of fibroblasts decreased correspondingly. Conclusion The skeletal muscle contusion in rats, contusion area of peripheral blood-derived fibroblasts changes in the number of time has a certain regularity, is expected to be used as a reference to infer skeletal muscle contusion time.