| Purpose:The protective effect of electroacupuncture combined with vibration training on motor function of HIE rats was explained by observing the effects of neurobehavioral HIE,brain tissue morphology and skeletal muscle cell apoptosis in rats with ischemic hypoxic encephalopathy(HIE).To explore the mechanism of this combined therapy to regulate the signaling pathway of nuclear factor kB(NF-kB)in skeletal muscle and to provide experimental basis for clinical guidance of motor function rehabilitation in children with cerebral palsy(CP)in rats.Material and method:Selected 100 male healthy SD rats of 21 days old(weighing about 150±20 g).After 7 days of adaptive feeding with free water intake,they were divided into sham operation group,model group,electroacupuncture group,vibration group and combined group according to random digital table method.Then the rats in the sham operation group were separated from the right common carotid artery without any disposal and suture incision,and the rats in the other groups were prepared HIE the rat model by modified Rice.After 24 h after operation,the rats in each group were evaluated,and the rats in electroacupuncture group,vibration group and combined group were treated after removing the failed and dead rats.The rats in theelectroacupuncture group needled Dazhui point,Renzhong point,Baihui point every day,and then connected with electroacupuncture instrument to adjust the frequency of 7-10 Hz treatment.The treatment lasted 15 minutes once a day for 2 weeks;The vibration group put the rats in a vibration platform which with a frequency of 25 Hz and an amplitude of 1.5mm.The treatment lasted 15 minutes once a day for2 weeks.The treatment of rats in the combined group was electroacupuncture plus vibration group.The treatment lasted 15 minutes once a day for 2 weeks.The neurobehavioral evaluation of each group of rats was carried out after the experiment.The rats’ s right hind limb gastrocnemius tissue and brain tissue were obtained by abdominal anesthesia.Detection of brain tissue by HE method;Detection of NF-kB P65、Bcl-2 expression in gastrocnemius by immunohistochemical method;The gene expression of NF-kB p65 、 Bcl-2 、 Bax in gastrocnemius tissue was detected by PCR method.Results:1.The neurobehavioral scale of rats showed that there was no neurological deficit in the sham operation group.After modeling 24 h,the neurobehavioral score increased in each group,which was statistically significant compared with the sham operation group(P<0.05).Two weeks after treatment,the three groups were lower than the model group(P<0.05),compared with the electroacupuncture group(P<0.05)and the vibration group(P<0.05).Compared with the electroacupuncture group,the vibration group score,but there was no significant difference(P>0.05).2.HE staining of brain tissue in the injured area showed that there was no injury,the cells were arranged neatly and the structure was clear.After modeling,there were ischemic necrosis lesions in all groups except sham operation,and there were cell arrangement disorder and necrosis.After 2 weeks of intervention,the degree of cell necrosis in each treatment group was improved compared with that in the model group,the cell death in the brain injury area of the combined group was significantly reduced,and the cell arrangement was basically normal.But cell death is still more,structure arrangement is still chaotic.3.The content of NF-kB p65 protein in gastrocnemius was detected by immunohistochemistry:After the establishment of the HIE model,the NF-kB p65 protein in the gastrocnemius muscle of the model rats was increased compared with that in the sham operation group(P<0.05).Two weeks after treatment,the electroacupuncture combined with vibration training group decreased(P<0.05),and the NF-kB p65 protein content in the vibration intervention group was higher than that in the electroacupuncture intervention group(P>0.05).4.The content of Bcl-2 protein in gastrocnemius was detected by immunohistochemistry:After the establishment of the model,the Bcl-2 in gastrocnemius muscle of each group was lower than that of sham-operated rats(P<0.05).Two weeks after intervention,electrovibration combined with vibration therapy Bcl-2 protein values were higher than those of electroacupuncture and vibration therapy(P<0.05).Electroacupuncture group increased more than vibration group(P>0.05).5.The expression of gastrocnemius NF-kB p65 、 Bax in rats by PCR assay showed:After modeling,the gene expression Bcl-2 the model group was lower than that of the sham operation group.The Bcl-2 gene expression in the combined group,vibration group and electroacupuncture group was higher than that in the model group(P<0.05)after 2 weeks of treatment,and the Bcl-2 gene expression in the two groups treated alone was lower than that in the combined treatment group(P<0.05),in which the electroacupuncture treatment was higher than that in the vibration therapy(P>0.05).6.PCR detection of Bcl-2 gene expression in rat gastrocnemius:After modeling,the Bcl-2gene expression in the model group was lower than that in the sham operation group.The expression of Bcl-2 genes in combined group,vibration group and electroacupuncture group was higher than that in model group(P<0.05).The expression of Bcl-2 gene in the two groups was lower than that in the combined treatment group(P<0.05),and the expression Bcl-2electroacupuncture therapy was higher than that in vibration therapy(P>0.05).Conclusion:1.Electroacupuncture combined with vibration training can improve the motor function of HIE rats.After treatment,the Bederson score was significantly lower than that of the simple treatment group,and the area of ischemic necrotic brain tissue was improved significantly.2.Electroacupuncture combined with vibration training can inhibit the apoptosis of skeletal muscle cells and reduce muscle atrophy by regulating.The results show that electroacupuncture combined with vibration training can inhibit NF-kB signaling pathway,Bax expression,enhance Bcl-2 expression,and reduce skeletal muscle cell apoptosis by inhibiting the cascade reaction effect of NF-kB regulation.and this may be one of the mechanisms by which combination therapy improves CP motor dysfunction. |
