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骨骼肌挫伤修复过程中巨噬细胞的趋化机制

The Mechanism of Macrophage Chemotaxis during the Repair Process Following Contusion of Skeletal Muscle

  • 摘要:
      目的  探究巨噬细胞在挫伤骨骼肌修复过程中的趋化机制。
      方法  将66只ICR雄性小鼠随机分为对照组(C组,n=11)和骨骼肌挫伤组(S组,n=55)。骨骼肌挫伤后12 h、1 d、5 d、10 d和15 d分别取双侧腓肠肌。HE染色观察骨骼肌挫伤后修复过程中的形态学变化,荧光定量PCR检测骨骼肌挫伤后巨噬细胞和趋化因子的时空表达规律。
      结果  ① 骨骼肌挫伤后12 h,可见肌纤维坏死、肿胀和大量炎性细胞浸润。伤后第5天,可见少量再生肌纤维,伤后第10天再生肌纤维数量大大增加,伤后第15天虽有少量再生肌纤维,但肌纤维结构完整性较好,骨骼肌损伤修复基本完成。②与对照组相比,M1巨噬细胞标志物(CD68)在骨骼肌挫伤后显著增加,随后M2巨噬细胞标志物(CD206和CD163)显著增加。③骨骼肌挫伤后MCP-1及其受体CCR2 mRNA表达均显著增加(P < 0.01)。SDF-1 mRNA在挫伤后12 h、10 d和15 d表达均显著增加(P < 0.01),其受体CXCR4表达无显著变化。④骨骼肌挫伤后,MCP-1与CD68存在强相关,亦与其受体CCR2呈强相关,而SDF-1只与其受体CXCR4相关。
      结论  MCP-1/CCR2轴可能参与了挫伤骨骼肌修复过程中M1巨噬细胞的趋化。

     

    Abstract:
      Objective  The objective of this study is to explore the mechanism of macrophage chemotaxis during the repair process following contusion of skeletal muscle.
      Methods  66 ICR male mice were randomly divided into control group(C group, n=11) and muscle contusion group(S group, n=55).Their gastrocnemius muscles were harvested at the time points of 12 h, 1 d, 5 d, 10 d and 15 d post-injury. The changes in skeletal muscle morphology were assessed by HE staining. The gene expression of macrophage and chemokines was analyzed by real-time PCR.
      Results  ① On 12 h post-injury, cross sections of gastrocnemius muscles showed substantial fiber damage and inflammatory infiltrate.On day 5 post-injury, a small quantity of centronucleated myofibers were observed.On day 10 post-injury, central nucleation phenomenon became more pronounced.On day 15 post-injury, central nucleation almost disappeared.② The molecule marker of M1 macrophages(CD68) increased significantly post-injury, and the molecule marker of M2 macrophages(CD206 and CD163) increased significantly later.③ MCP-1 and CCR2 mRNA increased significantly post-injury(P < 0.01).SDF-1 mRNA increased significantly at 12 h, 10 d and 15 d (P < 0.01).However, CXCR4 mRNA did not change significantly post injury.④ The results showed that there was significant correlation between MCP-1 and CD68, and CCR2.In addition, there was significant correlation between SDF-1 and CXCR4.
      Conclusion  MCP-1/CCR2 may be involved in the chemotaxis of M1 macrophages during skeletal muscle regeneration post-injury.

     

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