School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong Province, China.
Clinical Research Center, Department of Orthopaedic, Panzhihua Central Hospital, Panzhihua, Sichuan Province, China.
J Immunol Res. 2024 Nov 5;2024:5556042. doi: 10.1155/2024/5556042. eCollection 2024.
Manual therapy (MT) has been widely used in China to treat local tissue inflammation for a long time. However, there is a lack of scientific evidence for using MT in anti-inflammatory therapy, and its anti-inflammatory mechanism needs further clarification. We utilized MT to treat cardiotoxin (CTX) injury-induced skeletal muscle inflammation in C57BL6/J mice. We analyzed the underlying mechanism by integrating single-cell RNA sequencing (scRNA-seq) with molecular techniques. Hematoxylin and eosin (H&E) and immunohistochemical (IHC) staining were used to assess skeletal muscle inflammation and muscle fiber cross-sectional area (CSA). scRNA-seq, immunofluorescence, and western blot were performed to determine cellular and molecular outcome changes. Compared with CTX injury-induced skeletal muscle inflammatory mice, MT intervention significantly reduced proinflammatory cytokines interleukin (IL)-1, IL-6, and tumor necrosis factor alpha (TNF-) expression levels; scRNA-seq detected that neutrophil numbers and activity were maximum proportions increased in injured skeletal muscle among macrophage, T cells, B cells, endothelial cells, fast muscle cells, fibroblasts, and skeletal muscle satellite cells; and S100A9 gene expression was supreme in neutrophils. However, after treatment with MT, S100A9 protein expression and the numbers and activity of Ly6g+/Mpo+ neutrophils were significantly inhibited, thus reducing the inflammatory cytokine levels and exerting an anti-inflammatory effect by early clearing neutrophils. MT can mitigate localized inflammation induced by injured skeletal muscle, achieved by decreasing S100A9 protein expression and clearing neutrophils in mice, which may help advance therapeutic strategies for skeletal muscle localized inflammation.
手法治疗（MT）在中国长期以来一直被广泛用于治疗局部组织炎症。然而，MT 在抗炎治疗中的应用缺乏科学证据，其抗炎机制需要进一步阐明。我们利用 MT 治疗 C57BL6/J 小鼠的心脏毒素（CTX）损伤诱导的骨骼肌炎症。我们通过整合单细胞 RNA 测序（scRNA-seq）与分子技术来分析潜在的机制。苏木精和伊红（H&E）和免疫组织化学（IHC）染色用于评估骨骼肌炎症和肌纤维横截面积（CSA）。scRNA-seq、免疫荧光和 Western blot 用于确定细胞和分子结果的变化。与 CTX 损伤诱导的骨骼肌炎症小鼠相比，MT 干预显著降低了促炎细胞因子白细胞介素（IL）-1、IL-6 和肿瘤坏死因子-α（TNF-α）的表达水平；scRNA-seq 检测到在受伤的骨骼肌中，巨噬细胞、T 细胞、B 细胞、内皮细胞、快肌细胞、成纤维细胞和骨骼肌卫星细胞中，中性粒细胞的数量和活性增加比例最大；S100A9 基因表达在中性粒细胞中最高。然而，经 MT 治疗后，S100A9 蛋白表达以及 Ly6g+/Mpo+中性粒细胞的数量和活性均显著受到抑制，从而降低了炎症细胞因子水平，通过早期清除中性粒细胞发挥抗炎作用。MT 可以减轻受伤骨骼肌引起的局部炎症，其机制可能是通过降低 S100A9 蛋白表达和清除小鼠中的中性粒细胞，这可能有助于推进骨骼肌局部炎症的治疗策略。
