小鼠骨骼肌挫伤修复过程中肌再生因子、炎症因子及趋化因子的表达规律研究

发布时间：2018-03-07 19:17

本文选题：骨骼肌钝挫伤　切入点：损伤修复　出处：《上海体育学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过观察小鼠骨骼肌挫伤修复过程中肌再生因子、炎症因子以及相关趋化因子的表达规律,探讨肌再生因子、炎症因子以及相关趋化因子在骨骼肌挫伤修复过程中的表达规律以及可能发挥的作用。方法:共使用40只雄性C57小鼠,随机选择8只作为对照组(C组,n=8),其余小鼠进行骨骼肌钝挫伤(S组,n=32),损伤后分别在1,3,7,14进行小鼠腓肠肌取材。小鼠右侧腓肠肌制作石蜡标本,用于HE染色法显示骨骼肌损伤以及损伤修复的骨骼肌组织形态结构,左侧腓肠肌用于基因表达检测,用荧光定量PCR检测肌再生调节因子(MyoD,myogenin,IGF-1,MGF,HGF,TGF-β1,Myostain),炎症因子(IL-1β,IL-6,IL-10)以及趋化因子(CCL2,CCL3,CCL5,CCL8,CXCL9,CXCL10,CXCL12)mRNA 表达变化。结果:1、骨骼肌钝挫伤后第3天有少量再生肌纤维,第7天显著增加,第14天基本恢复正常。2、MyoD和Myogenin分别是肌卫星细胞增殖和分化特异性标志物,在骨骼肌钝挫伤后显著表达(P0.01)。3、M1型巨噬细胞标志物CD68 mRNA在骨骼肌损伤的第1天和第3天显著增加(p0.01),M2型巨噬细胞标志物CD163和CD206 mRNA都在骨骼肌损伤的第1天显著增加(p0.01),之后CD163 mRNA迅速下降,而CD206 mRNA在骨骼肌损伤后第三天继续增加(p0.01)4、与对照组相比,生肌调节因子(IGF-1,MGF,HGF,TGF-β 1)mRNA在损伤后表达显著增加(P0.05或P0.01),MSTNmRNA在损伤后再生阶段的第7天到第14天,MSTN mRNA表达显著低于对照组(P0.01)。5、炎性细胞因子(IL-1β,IL-6,IL-10)mRNA 在骨骼肌损伤后显著性表达(p0.01或P0.05)6、趋化因子(CCL2,CCL3,CCL5,CCL8,CXCL9,CXCL10,CXCL12)mRNA 在骨骼肌钝挫伤后显著性表达,其中CCL2 mRNA在骨骼肌损伤后第1天已经开始显著性升高(P0.01),CCL3,CCL5与CCL8 mRNA在骨骼肌损伤后第1天显著性增加(p0.01),第7天与第3天相比,mRNA水平虽有下降,但与对照组相比,仍显著增加(p0.01)。CXCL9,CXCL12 mRNA均在第7天显著性增加(p0.01),随后下降。其中CXCL10 mRNA在损伤后第一天呈增加趋势,损伤后第3天与对照组相比呈显著性差异,这种增加趋势直到损伤后第7天后才开始下降。结论:1、骨骼肌挫伤后,M1、M2型巨噬细胞标志物及多种炎性因子(IL-1β,IL-6,IL-10)表达上调,它们可能在骨骼肌损伤修复过程中发挥重要作用。2、趋化因子CCL2,CXCL10可能参与了骨骼肌挫伤修复阶段免疫细胞的趋化。3、骨骼肌挫伤修复过程中,IGF-1,MGF,HGF,TGF-β1,Myostain等肌再生调控因子可能参与了对骨骼肌再生的调控。
[Abstract]:Objective: to investigate the expression of muscle regeneration factors, inflammatory factors and related chemokines during the repair of skeletal muscle contusion in mice. Expression of inflammatory factors and related chemokines in skeletal muscle contusion repair and their possible role. Methods: 40 male C57 mice were used. Eight mice were randomly selected as control group C, and the rest mice were treated with skeletal muscle contusion group S, and then the gastrocnemius muscle samples were taken from the gastrocnemius muscle at 1: 3 and 714, respectively. Paraffin wax specimens were made from the right gastrocnemius muscle of the mice. The left gastrocnemius muscle was used to detect gene expression. Fluorescence quantitative PCR was used to detect the mRNA expression of myogenic regulatory factor MyoDmyogenin IGF-1MGF- 尾 1 (Myostaina), the inflammatory factor IL-1 尾 -IL-6 IL-10) and the chemokine CCL2CCL5CCL5CCL8CXCL9CXCL10CXCL1212. Results: there was a small amount of regenerated muscle fibers on the 3rd day after blunt contusion of skeletal muscle, which was significantly increased on the 7th day. On the 14th day, MyoD and Myogenin were respectively specific markers for proliferation and differentiation of myosatellous cells. After skeletal muscle contusion, the expression of CD68 mRNA, a type M 1 macrophage marker, was significantly increased on the 1st and 3rd day after skeletal muscle injury. Both CD163 and CD206 mRNA were significantly increased on the first day of skeletal muscle injury, and the expression of CD206 mRNA increased significantly on the first day of skeletal muscle injury. Then CD163 mRNA dropped rapidly, However, CD206 mRNA continued to increase on the third day after skeletal muscle injury, compared with the control group. The expression of MSTN mRNA of IGF-1 / MGF- HGF- 尾 _ (1) increased significantly after injury. The expression of MSTN mRNA was significantly lower than that of control group from 7th to 14th day after injury, and the expression of inflammatory cytokine IL-1 尾 IL-6IL-10 mRNA was significantly higher than that of control group (P 0.01 or P 0.01 or P 0.01 or P 0.01 or P 0.01) after the injury of the skeletal muscle. The expression of MSTN mRNA was significantly lower than that of the control group from day 7 to day 14. The expression of the inflammatory cytokine IL-1 尾 IL-6mGF- 尾 IL-10 mRNA was significantly higher than that of the control group (P 0.01 or P 0.01). The expression of CXCL10, CXCL10, CXCL12 mRNA in skeletal muscle after blunt contusion was significant. CCL2 mRNA began to increase significantly on the first day after skeletal muscle injury. P0.01CCL5 and CCL8 mRNA increased p0.01a on the 1st day after skeletal muscle injury, and decreased on the 7th day compared with the 3rd day, but compared with the control group. The mRNA of CXCL9 + CXCL12 was significantly increased on the 7th day, and then decreased. The CXCL10 mRNA increased on the first day after injury, and on the third day after injury, there was a significant difference compared with the control group. This increased trend did not begin to decrease until 7 days after injury. Conclusion the expression of M1M2-type macrophage markers and various inflammatory cytokines (IL-1 尾 / IL-6 / IL-10) is up-regulated after skeletal muscle contusion. They may play an important role in the repair of skeletal muscle injury. The chemokine CCL2CXCL10 may be involved in the chemotaxis of immune cells in the repair stage of skeletal muscle contusion. IGF-1, MGFHGF- 尾 1, Myostain and other muscle regeneration regulatory factors may be involved in the repair of skeletal muscle contusion. Participate in the regulation of skeletal muscle regeneration.
【学位授予单位】：上海体育学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：G804.2

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