推拿疗法对骨骼肌纤维化大鼠MMP-1和TIMP-1表达的影响

Effect of massage therapy on the expression of MMP-1 and TIMP-1 in rats with skeletal muscle fibrosis

关键词:  推拿  骨骼肌  纤维化  MMP-1  TIMP-1

Key words:  massage  skeletal muscle  fibrosis  MMPs  TIMP-1

摘要:

[目的] 观察推拿疗法对大鼠急性腓肠肌钝挫伤后纤维修复的影响，探讨其可能的作用机制。[方法] 将30只雄性SD大鼠随机分为对照组、模型组、推拿治疗组（n=10），运用自制砸伤装置复制经典骨骼肌钝挫伤模型。治疗组进行推拿治疗，每次干预10 min，每日1次，连续实施25 d。通过苏木精-伊红（HE）染色观察大鼠腓肠肌纤维化程度；qPCR和蛋白印迹检测大鼠腓肠肌基质金属蛋白酶-1（MMP-1）、金属蛋白酶组织抑制因子-1（TIMP-1）蛋白的表达情况。[结果] HE染色镜下发现对照组大鼠骨骼肌纤维排列整齐，形态结构完整；模型组大鼠骨骼肌纤维排列紊乱，甚至断裂，肌成纤维细胞增多，细胞外基质ECM）增多且胶原纤维明显沉积；治疗组大鼠骨骼肌纤维排列较整齐、间距小，且大小均一，胶原纤维生成减少，仅有少量ECM生成，损伤基本恢复。qPCR和蛋白印迹结果显示模型组大鼠损伤腓肠肌MMP-1的mRNA和蛋白表达量及MMP-1/TIMP-1比值明显降低（P<0.01），而TIMP-1的mRNA和蛋白表达水平明显升高；给予推拿治疗后可明显提高损伤大鼠腓肠肌MMP-1的mRNA和蛋白水平及MMP-1/TIMP-1的比值（P<0.01，P<0.05），且降低TIMP-1的转录水平（P<0.01，P<0.05）。[结论] 推拿治疗可有效减轻骨骼肌钝挫伤后纤维化程度，其作用可能与上调MMP-1/TIMP-1比值有关。

Abstract:

[Objective] To observe the effect of massage therapy on fiber repair in rats with acute gastrocnemius contusion,explore its possible mechanism of action. [Methods] Thirty male SD rats were randomly divided into control group,model group and massage treatment group (n=10),the classic skeletal muscle blunt trauma model was replicated using a self-made bruising device. Treatment group for massage treatment,ten min per intervention,once a day,25 days in a row. The degree of rat gastrocnemius fibrosis was observed by HE staining;qPCR and Western blot were used to detect the expression of MMP-1 and TIMP-1 protein in rat gastrocnemius muscle. [Results] Under the HE staining microscope,the skeletal muscle fibers of the control group were arranged neatly,complete morphological structure;disorder of skeletal muscle fibers in the model group,even broken,increased myofibroblasts,increased ECM and significant deposition of collagen fibers;the skeletal muscle fibers of the treatment group were arranged neatly,small spacing,and uniform in size,erduced collagen fiber production,only a small amount of ECM generation,the damage is basically restored. qPCR and Western blot results showed that the mRNA and protein expression of MMP-1 and the ratio of MMP-1/TIMP-1 in the gastrocnemius muscle of the model group were significantly decreased (P<0.01),TIMP-1 mRNA and protein expression levels were significantly increased;After giving massage treatment,can significantly improve the mRNA and protein levels of MMP-1 in the gastrocnemius of injured rats,and the ratio of MMP-1/TIMP-1 (P<0.01,P<0.05),and the transcription level of TIMP-1 was decreased (P<0.01,P<0.05). [Conclusion] Massage treatment can effectively reduce the degree of fibrosis after skeletal muscle blunt trauma,it's effect may be related to up-regulation of the MMP-1/TIMP-1 ratio.