浮针对骨骼肌挫伤小鼠肌肉恢复的影响

Effect of Fu's subcutaneous needling on muscle recovery in mice with skeletal muscle contusion

关键词:  骨骼肌挫伤  纤维化  炎症  巨噬细胞  浮针  针灸  疼痛  小鼠

Key words:  skeletal muscle contusion  fibrosis  inflammation  macrophage  Fu's subcutaneous needling  acupuncture  pain  mice

摘要:

目的探讨浮针对骨骼肌挫伤的治疗效果和相关机制。 方法所有C57/BL6雄性小鼠随机分为4组: 对照组（control group, C组），模型组（model group, M组），浮针组（Fu's subcutaneous needling group, FSN组）和针灸组（acupuncture group, ACP组）。除了对照组，其他小鼠的左侧腓肠肌被挫伤损伤。FSN组每2d治疗1次，配合再灌注方法，一共6次，分别为第1、3、5、7、9、11天。ACP组的治疗时间与FSN组相同。通过von Fery和Hargreaves实验评估痛觉过敏的改变,通过测量两侧腓肠肌湿重和周长评估肌肉水肿程度，通过组织学和RT-qPCR评估炎症和纤维化，使用流式细胞术评估巨噬细胞亚群。 结果FSN组和ACP组的热痛反射潜伏期明显延长。浮针干预明显减轻损伤肌肉的水肿程度，针灸治疗无明显影响。FSN治疗在第3天降低了炎症因子和浸润白细胞的基因表达。同时，在第7天和第14天，它减少了受损肌肉的纤维化面积。FSN组在第7天，M1巨噬细胞的数量减少，而M2巨噬细胞的数量增加。ACP组巨噬细胞亚群变化不明显。 结论骨骼肌挫伤后FSN治疗不仅可以缓解热痛觉过敏，减轻肌肉损伤中的早期炎症水平和肌肉水肿，还可以减少纤维化。这些可以通过促进M2巨噬细胞极化来实现。

Abstract:

ObjectiveTo investigate the therapeutic effect and related mechanisms of Fu's subcutaneous needling(FSN) for skeletal muscle contusion in mice. MethodsC57/BL6 male mice were randomly divided into 4 groups: control group, model group, FSN group and acupuncture（ACP） group. Except for control group, the left gastrocnemius muscle of mice was injured by contusion. The FSN group received FSN treatment plus reperfusion once every alternative day for 6 times; the ACP group received acupuncture at the acupoint Zusanli(ST 36). The changes in hyperalgesia were evaluated by von Fery test and Hargreaves test; the degree of muscle edema was assessed by measuring the wet weight and circumference of gastrocnemius muscles; inflammation and fibrosis were evaluated by histology and RT-qPCR; and subpopulations of macrophage were evaluated using flow cytometry. ResultsThe thermal withdrawal latency significantly prolonged in FSN and ACP group. FSN treatment significantly reduced the degree of muscle edema, and acupuncture treatment has no effect. FSN treatment reduced the gene expression of inflammatory factors and infiltrating leukocytes on the 3 rd day. Meanwhile, it decreased the fibrotic area of damaged muscles on 7th day and 14th day. The number of M1 macrophages decreased, while the number of M2 macrophages increased after FSN treatment on 7th day. The macrophage subgroup in the acupuncture did not change significantly. ConclusionThe study suggests that FSN treatment not only alleviate thermal hyperalgesia and reduce early inflammation levels and muscle edema, but also reduce fibrosis in mice with skeletal muscle contusion, which may be achieved by promoting M2 macrophage polarization.