大鼠挫伤骨骼肌中M1/M2型巨噬细胞时间相关性研究

Study on time correlation of M1/M2 macrophages in skeletal muscle contusion of rats

目的研究大鼠骨骼肌挫伤后M1/M2型巨噬细胞浸润的动态变化规律结合组织学改变建立损伤时间推断方法。方法建立大鼠骨骼肌挫伤模型,挫伤后6h、12h、24h、36h、48h、60h、72h、96h、144h、216h、336h为实验组,对照组不进行任何处理。采用HE染色法和免疫荧光多重染色方法观察骨骼肌挫伤后修复规律,运用全景组织细胞定量分析系统检测损伤区内单位面积细胞核数量、M1及M2型巨噬细胞数量和比例变化规律并建立与损伤时间的关系。结果在0.5mm×0.5mm范围内,M1型巨噬细胞比例在0.05±0.02~0.18±0.12且M2型巨噬细胞比例在0.02±0.01~0.04±0.02范围内时,推断该组织的损伤时间为伤后6h~48h;M1型巨噬细胞比例0.22±0.02且M2型巨噬细胞比例在0.09±0.02范围内时,推断该组织的损伤时间为损伤后60h;M1型巨噬细胞比例0.1±0.02且M2型巨噬细胞比例在0.16±0.07范围内时,推断该组织的损伤时间为损伤后144h;M1型巨噬细胞比例0.05±0.01~0.09±0.02且M2型巨噬细胞比例在0.07±0.02~0.09±0.01范围内时,推断该组织的损伤时间为损伤后216h~336h。结论大鼠挫伤骨骼肌中M1/M2型巨噬细胞时间相关性表达规律可作为损伤时间推断的新依据,结合病理组织学改变可进一步提高损伤时间推断的准确性。

Objective To study the dynamic changes of M1/M2 macrophage infiltration after skeletal muscle contusion in rats, and to establish a method of injury time estimation based on histological changes. Methods Rat skeletal muscle contusion model was established. 6 h、12 h、24 h、36 h、48 h、60 h、72 h、96 h、144 h、216 h and 336 h after contusion were treated as experimental groups, while the control group received no treatment. HE staining method and immunofluorescence multiple staining method were used to observe the repair process of skeletal muscle after contusion, and panoramic tissue cell quantitative analysis system was used to detect the change of nucleus number per unit area, M1 and M2 type macrophage number and proportion, and establish the relationship with the injury time. Results When the ratio of M1-type macrophage was within the range of 0.05±0.02~0.18±0.12 and the M2-type macrophage was 0.02±0.01~0.04±0.02 in the area of 0.5 mm×0.5 mm, it was inferred that the injury time was 6 h-48 h. When the ratio of M1-type macrophage was 0.22±0.02 and the M2-type macrophage was 0.09±0.02, it was inferred 60 h after injury.When the ratio of M1-type macrophage was 0.1±0.02 and the ratio of M2-type macrophage was 0.16±0.07, it was inferred that the injury time of the tissue was 144 h.When the M1-type macrophage ratio was within the range of 0.07±0.02~0.09± 0.02 and the M2-type macrophage ratio was within the range of 0.07±0.02~0.09±0.01, it was inferred that the injury time of the tissue was 216 h~336 h. Conclusion The time-dependent expression of M1/M2 macrophages in rat contused skeletal muscle can be used as a new basis for the prediction of injury time, and the accuracyof estimation of injury time can be further improved by combining with histopathological changes.