基于Wnt/β-catenin信号通路探讨推法对骨骼肌过劳性损伤大鼠的影响及作用机制  

Effect and Mechanism of Massage Manipulation on Rats with Skeletal Muscle Overuse Injury Based on Wnt/β-catenin Signaling Pathway

作　　者：段苗苗 黄博 阮磊 穆盼盼 陈亦民 彭亮[1] DUAN Miaomiao;HUANG Bo;RUAN Lei;MU Panpan;CHEN Yiming;PENG Liang(Acupuncture and Moxibustion and Rehabilitation College,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China)

机构地区：[1]湖南中医药大学针灸推拿与康复学院,湖南长沙410208

出　　处：《康复学报》2024年第6期617-624,共8页Rehabilitation Medicine

基　　金：国家自然科学基金项目(82174521)。

摘　　要：目的观察推法对骨骼肌过劳性损伤大鼠修复再生的影响,基于Wnt/β-catenin信号通路探讨其作用机制。方法选择42只SD大鼠按随机数字表法分为空白组、模型组和干预组,每组14只。除空白组外,模型组、干预组参考Armstong离心运动方案训练建立大鼠骨骼肌过劳性损伤模型。空白组、模型组右后肢腓肠肌及相邻肌肉部位涂抹医用级凡士林,但不予推法干预;干预组右后肢腓肠肌部位进行推法干预(压力约为2 N、频率80次/min,3 min/次,每天上午、下午各干预1次,共持续3 d)。分别于造模结束后24 h和干预结束后24 h采用平衡木测试评估大鼠骨骼肌损伤后的运动功能;苏木精-伊红染色法(HE)观察腓肠肌的病理学改变;采用EM-1400型透射电子显微镜观察骨骼肌纤维超微结构;采用荧光定量PCR仪检测肌源性调节因子成肌分化抗原(MyoD)、肌肉生成素(MyoG)和配对盒转录基因7(Pax7)mRNA转录水平;采用Western blot法检测MyoD、MyoG、Pax7、Wnt3a、Axin2和β-catenin蛋白表达水平。结果(1)运动功能:与空白组比较,模型组、干预组平衡木通过时间、滑爪次数均明显增加,差异均具有统计学意义(P<0.05);与模型组比较,干预组干预后平衡木通过时间、滑爪次数均明显减少,差异均具有统计学意义(P<0.05)。(2)腓肠肌组织病理学改变:模型组腓肠肌内大量的肌细胞内可见裂纹,肌原纤维断裂;与模型组比较,干预组腓肠肌组织损伤程度减轻,可见肌纤维形态规则,排列整齐。(3)腓肠肌组织超微结构改变:模型组局部肌原纤维束断裂、溶解,少量Z线结构溶解、模糊;与模型组比较,干预组腓肠肌细胞轻微水肿,肌原纤维束结构清晰,部分肌原纤维束断裂,Z线结构清晰、排列较整齐。(4)腓肠肌组织MyoD、MyoG、Pax7 mRNA转录水平:与空白组比较,模型组和干预组MyoD、MyoG、Pax7 mRNA转录水平均明显更高(P<0.05);与模型组比较,干预组MyoD、MyoGObjective To observe the effect and its mechanism of massage manipulation on the repair and regeneration of skeletal muscle overuse injury in rats based on Wnt/β-catenin signaling pathway.Methods A total of 42 SD rats were randomly divided into blank group,model group and intervention group,with 14 cases in each group.Armstong centrifugal exercise program was used to establish the rat skeletal muscle overuse injury model in the model group and the intervention group,except for the blank group.Medical vaseline was smeared on the right hindlimb gastrocnemius muscle and adjacent muscles in the blank group and the model group,but no massage manipulation was given.Massage manipulation was given at the right hind gastrocnemius muscle in the intervention group,with the pressure of about 2 N,the frequency of 80 beats per minute,three minutes a time,once in the morning and once in the afternoon,lasting for three days.At 24 h after modeling and 24 h after intervention,the balance beam test was used to assess the motor function of rats with skeletal muscle injury;hematoxylin-eosin(HE)staining method was used to observe the pathological changes of the gastrocnemius muscle;an EM-1400 transmission electron microscope was used to observe the ultrastructure of the skeletal muscle fibers;fluorescence PCR was used to detect the myogenic regulatory factors including myogenic differentiation antigen(MyoD),myogenin(MyoG)and paired box gene 7(Pax7)mRNA transcript levels;Western blot was used to detect MyoD,MyoG,Pax7,Wnt3a,Axin2 andβ-catenin protein expression levels.Results(1)Motor function:compared with the blank group,the time to pass the balance beam and the number of paw slips in the model group and the intervention group increased significantly,and the differences were statistically significant(P<0.05).Compared with the model group,the time to pass the balance beam and the number of paw slips in the intervention group decreased significantly after intervention,and the differences were statistically significant(P<0.05).(2)Histop

关 键 词：过劳性损伤 骨骼肌 修复再生 运动功能 推法 WNT/Β-CATENIN信号通路 

分 类 号：R73[医药卫生—肿瘤]