推拿疗法对骨骼肌纤维化大鼠MMP-1和TIMP-1表达的影响

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。

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

Objective] To investigate the pathogenesis of the rat model of coronary heart disease with phlegm and blood-stasis syndrome based on PI3K/Akt pathway.Methods] Wistar male rats were randomly divided into blank group,sham operation group,turbidity group,blood stasis group and phlegm and blood-stasis group. On the 54th day,the left anterior descending coronary artery of the rats in blood-stasis group and the phlegm and blood-stasis group were ligated,and the rats in the sham operation group only threaded without ligation. The rats in the blank group,the blood-stasis group and the sham operation group were fed with normal feed for 8 weeks,while the phlegm group and the phlegm and blood-stasis group were fed with high fat diet for 8 weeks. The apoptosis of cardiomyocytes was observed by Tunel staining. The expressions of Akt,p-Akt,Bax,Bcl-2 and Caspase-3 were detected by Western blot.Results] The results of Tunel staining showed that the apoptosis of phlegm group,blood-stasis group and phlegm and blood-stasis group increased significantly. Western blot results showed that the expression levels of Akt,p-Akt and Bcl-2 protein in blood-stasis group and phlegm and blood-stasis group were significantly lower than those in the blank group (P<0.05 or P<0.01). The expression levels of Bax and Caspase-3 were significantly increased (P<0.05 or P<0.01). The expression of p-Akt protein in the phlegm group was significantly increased (P<0.05),and there was no significant difference in the expression of Akt,Bcl-2,Bax and Caspase-3.Conclusion] It is suggested that PI3K/Akt pathway maybe one of the pathogenesis of coronary heart disease with phlegm and blood-stasis syndrome by down-regulating Akt,p-Akt and Bcl-2 proteins in cardiomyocytes,up-regulating Bax and Caspase-3 proteins.