| Object:By detecting the serum CK and CK-MM levels and the expression of Pax7,NICD,RBPJ-k,COLV,and CalcR proteins in the gastrocnemius muscle of rats,the effect of massage on the injury intervention of EIMD rats was observed,and the mechanism of Notch/CollagenV/CalcR axis in the intervention of MSC microenvironment in EIMD model rats by massage was explored.Methods:72 male Sprague-Dawley rats were randomly divided into group C(blank control group),group E(simple model group)and group T(model massage group).They were divided into immediate group,12h,24h and 48h according to the sampling time.The model was established by "disposable high-load centrifugal exercise".Group T was given massage immediately after model establishment,and another massage intervention was given in the corresponding time phase(parameters:strength 400 ± 50g,10 min/time,60 times/min).The general condition of rats was observed,histomorphology was observed by HE staining,ultrastructure was observed by electron microscopy,serum CK and CK-MM content levels were measured by ELISA,and Pax7,NICD,RBPJ-K,COLV,and CalcR protein expression in muscle tissue was detected by Western blot.SPSS 19.0 statistical software was used for data processing.Results:1.General observation:The rats in the model group(E group,T group)had decreased motor ability,uneven breathing,and swelling of some limbs.After the end of modeling,the motor ability was recovered,and T group was superior to E group.2.Hi stomorphol ogi cal observation:Immediately to 48 hours,myofibrils were disorganized and inflammatory cell infiltration was gradually aggravated in group E and peaked at 48 hours,myofibrils were disorganized and inflammatory cell infiltration was observed in group T and peaked at 24 hours,and the recovery of all phases in group T was better than that in group E.3.Ultrastructure observation:Immediately to 48 hours,myofibrils were disorganized and mitochondrial swelling was gradually aggravated in group E,peaking at 48 hours,myofibrils were disorganized and inflammatory cells were infiltrated in group T,peaking at 24 hours,and the recovery of all phases in group T was better than that in group E.4.CK and CK-MM levels:The serum CK and CK-MM levels of rats in groups E and T reached the peak value immediately,gradually decreased with time,and approached the level of group C by 48 hours,and decreased significantly in group T compared with group E.5.Pax7 expression:(1)Intragroup analysis:Immediately to 48 hours,the expression was stable in group C,gradually increased in groups E and T,and was highest by 48 hours.(2)Between-group analysis:The expression of Pax7 in E and T groups was significantly higher than that in C group,and that in T group was higher than that in E group.There were significant differences among C,E and T groups(P ?0.05).6.Notch/CollagenV/CalcR axis related protein expression results:(1)NICD expression:?Intragroup analysis:Immediately to 48 hours,NICD expression was slightly decreased in group C,peaked immediately in groups E and T,and then decreased,tended to be stable at 24 hours in group E,and still decreased at 48 hours in group T.?Between-group analysis:The expression of NICD in group E and T was significantly higher than that in group C,and group E ranked first.At immediate,12 h,and 24 h,there was a significant difference in C,E,and T(P ?0.05);at 48 h,there was a significant difference in the increase between groups E and T compared with group C(P ?0.05),and there was no significant difference in the decrease between group T and group E(P?0.05).(2)RBPJ-K expression:?Analysis within the group:Immediately to 48 hours,RBPJ-K expression increased slightly and then decreased in group C,peaked immediately in groups E and T,then decreased,tended to be stable at 24 hours,and slightly decreased at 48 hours.?Between-group analysis:The expression of protein in each phase of group E and T was significantly higher than that of group C,and group E ranked first.Immediately,compared with group C,the increase in group E was significantly different(P ?0.05),the increase in group T was not significantly different(P? 0.05),compared with group E,the decrease in group T was not significantly different(P? 0.05);at 12h,24h,and 48 h,the differences in C,E,and T were significant(P ?0.05).(3)COLV expression:?Intragroup analysis:Immediately to 48 hours,collagenV expression decreased first and then increased in group C,decreased significantly immediately to 12 hours and then increased in group E,and slightly decreased and then increased immediately to 12 hours in group T,all peaking at 48 hours.?Between-group analysis:CollagenV expression was significantly higher in group C than in groups E and T,and higher in group T than in group E.Immediately,compared with group C,group E and T decreased significantly(P ?0.05),compared with group E,group T increased without significant difference(P>0.05);at 12h and 24h,compared with group C,group E and T decreased significantly(P ?0.05),compared with group E,group T increased significantly(P ?0.05);at 48h,compared with group C,group E decreased significantly(P ?0.05),T decreased slightly,without significant difference(P?0.05);compared with group E,group T increased significantly(P ?0.05).(4)CalcR expression:?Intragroup analysis:Immediately to 48 hours,CalcR expression increased steadily and slightly in group C,increased first and then decreased in group E immediately to 12 hours,and continued to decrease in group T.?Between-group analysis:CalcR expression was significantly higher in group E than in group C,higher in group E than in group T immediately,12 hours,and 24 hours,and higher in group T than in group E at 48 hours.Immediately,12h and 24h,compared with group C,group E and T had significant increase(P ?0.05),compared with group E,group T had no significant increase(P? 0.05);at 48h,compared with group C,group E had significant increase(P ?0.05),T had slight decrease,without significant difference(P?0.05);compared with group E,group T had significant difference(P ?0.05).Conclusion:(1)Massage can promote the recovery of myofibril arrangement,reduce the degree of mitochondrial swelling,down-regulate serum CK and CK-MM to normal levels,and effectively improve the symptoms of injury in a rat model of EIMD induced by a single high-load eccentric exercise.(2)Pax7 expression increased after skeletal muscle injury in rats,suggesting that MSC initiated proliferation,and the massage group was more significant and lasted until 48 hours,and massage promoted MSC activation and proliferation.(3)During the intervention of EIMD by massage,Notch/CollagenV/CalcR axis maintains the microenvironment state,maintains MSC function,promotes the repair and regeneration of damaged skeletal muscle fibers,and maintains the normal structure and functional stability of skeletal muscle cells by regulating the expression of NICD,RBPJ-K,COLV,and CalcR at different phases in the microenvironment of muscle stem cells. |
PDF Full Text Request