The Effect Of Electroacupuncture At The Middle Point Of The Acupoint On The Autophagy-related Proteins Beclin1, P62 During The Repair Of Lumbar Multi-fissure Injury In Model Rats

BackgroundAt the waist,the multifidus is located in the deep part of the erector spinae.Its function is to stabilize the lumbar spine,ensure the close connection of each lumbar spine,and finely distribute the pressure on the lumbar spine.The damage,dysfunction and atrophy,of the lumbar multifidus can destroy the stability of the lumbar spine and cause back pain.According to surveys,2/3 of the people in Western countries are affected by low back pain,and disability caused by low back pain is even higher in the world,bringing pain to millions of families.Therefore,it is particularly so important to promote the repair and functional reconstruction of the multifidus muscle for the treatment of acute and chronic low back pain and improve the quality of life of patients with low back pain.It will also provide stronger evidence for the treatment of lumbago by acupuncture.The unique therapeutic effect of acupuncture at the "Weizhong"(BL40)Point on low back pain has been highly recognized by the majority of scholars.However,the research on the mechanism of action of Weizhong was focused on the neural sources,analgesia,and microcirculation changes in Weizhong in the past.There is no good research evidence to promote the regeneration and repair of skeletal muscle injury yet.After muscle injury,Activation of muscle satellite cells is a prerequisite for muscle repair,and its activation requires the participation of autophagy after the muscle injuried.So the level of autophagy directly affects the quality and time of repair.On the one hand,this study intends to observe the multifidus muscles from histomorphology;on the other hand,to observe the effect of electroacupuncture "Weizhong" on the expression of autophagy-related proteins Beclinl and p62 to explore the mechanism of electroacupuncture "Weizhong" promotes the repair of multifidus muscle injury.ObjectiveTo observe the recovery of rat model of lumbar multifidus muscle injury caused by bupivacaine from the perspective of morphology,and compare withMethodsExperiment 1:Lumbar multifidus muscle damage was observed in rat model.A total of 54rats were randomly divided into blank group,experiment group and experiment with LY294002 group,24 rats in each group.Both sides of multifidus muscle(L4 and L5)in experiment group and experiment with LY294002 group were injected with 0.5%bupivacaine(0.5%,350mg/kg),the needle(No.4th)is close to the spine to enter the muscle until the contact with the articular process and the mastoid bone surface back casing 1mm no blood,indicating that the needle has reached the 100μl,each point of the injection of bupivacaine,respectively,the time must be longer than 3 seconds,to make sure the absorption of drugs.The rats in experiment group accepted injection with NS once day,and the rats in experiment with LY294002 group accepted injection with LY294002 once day..The blank group will not take any measure.Experiment 272 Healthy adult male SD rats,body mass 280～320g,random cage feeding,adaptive feeding for 7 days,All these rats were stratified by body mass,the random number table method was divided into blank group,model group,and EA"Weizhong"(BL 40)group,each group of 24,each group of rats were randomly divided into 1 days,3 days,7 days three time points,8 rats per time point.The modeling method with the experimental 1.The rats in BL40 electroacupunture group accepted electroacupunture once day after model establishment,the rats in model group were catched and fixed at same time.Injury rats were observed after HE staining by light microscopy multifidus Histological changes on daysl,3,7,The expression of Beclin1,p62 was measured by immunohistochemical method and western-blot method.The electro acupuncture Weizhong,autophagy,multifidus muscle injury as the starting point to further explore the acupuncture Weizhong promote the multifidus muscle injury repair mechanism.Results1.The multifidus muscle tissue of different time points under HE staining,Masson staining,Iron Hematoxylin Staining were changed significantly except the 1th day after modeling,the two groups of the multifidus muscle showed similar histological morphology,while the recovery of skeletal musclethe in the other 3rd and 7 days were better than the experiment with LY294002 group.2.The effects of electroacupuncture at BL40 on the expression of Beclinland p62 after the rat,s multifidus muscle were injuried by the injection of Bupivacaine.The expression of Beclinl at day 1,3,7 ranked as model group was higher than blank group(P<0.01),while ranked as BL40 electroacupunture group higher than model group at day 3 and day 7(P<0.05);The model group-s expression of p62 was higher than blank group at day 3 and day 7(P<0.05);Compared with the model group,the expression of p62 in BL40 electroacupunture group decreased at day 1,day 3 and day7(P<0.05).Conclusion1.Observation of damaged multifidus by three staining methods,we found:Autophagy pathway inhibitor LY294002 can specifically prevent autophagy,the delay the repair of skeletal muscle damage,also shows autophagy can promote the repair of skeletal muscle damage.2.EA "Weizhong"(BL 40)can increase the expression of autophagy associated protein Beclinl,improve autophagy levels and decrease the expression of P62,so as to be beneficial to skeletal muscle repair.