Effects Of Acupotomy Therapy On PI3K/Akt Signaling Pathway Of Posterior Cervical Extensor Muscles Cells In Cervical Spondylosis Rabbits

Objective:Cervical spondylosis(CS)is a common public health problem worldwide.It has a high disability rate and brings huge pressure to individuals,families,medical system and social economy.In recent years,acupotomy for the treatment of cervical spondylosis due to significant curative effect,convenient operation,labor saving and other advantages has been widely concerned by the community.Evidence-based medicine systematic evaluation and Meta analysis showed that acupotomy was safe and reliable in the treatment of cervical spondylosis.Also,literature metrology studies showed that cervical spondylosis was the dominant disease and indication of acupotomy.However,the basic research on the treatment of cervical spondylosis with acupotomy is weak.this study to simulate the clinical characteristics of the onset of cervical spondylosis,established a rabbit model of cervical spondylosis,observed the histological and ultrastructural changes of the posterior cervical muscle with light microscopy and transmission electron microscopy.And to investigate the effect of acupotomy intervention on the PI3K/Akt signaling pathway of the posterior cervical extensor muscles cells in cervical spondylosis rabbits model,elaborating the possible effect mechanism of acupotomy in the treatment of cervical spondylosis,so as to provide experimental basis for the selection of acupotomy for clinical treatment of cervical spondylosis.Methods:1.Animals grouping: according to the random number table,40 New Zealand male rabbits were randomly divided into 4 groups(blank group,model group,acupotomy group and electro-acupuncture group)with 10 rabbits in each group;2.Establishment of cervical spondylosis rabbit model: except the blank group,the other three groups established animal models of cervical spondylosis by long-term bow position for 12 weeks.The success of the model was verified by cervical X-ray plain film.3.Treatment methods: after successful modeling,acupotomy was performed at the point of indurate,cord-like mass and spinous process,once a week for three weeks.For electro-acupuncture group,electro-acupuncture group was applied to bilateral “Tianzhu”(BL 10),“Jingbailao”(EX-HN 15),“Dazhu”(BL 11)of the rabbit neck muscle,once daily,three times a week for 3 weeks.The blank group and model group were fed normally.4.One week after the end of intervention,the posterior extensor muscles were taken from bilateral rabbits neck muscles.The changes of HE and ultrastructure of cervical muscle were observed under light microscope and electronic microscope respectively.5.The expression level of PI3 K gene was detected by Real-time PCR method,and the expression of PI3 K,Akt and P-Akt proteins were detected by Western blot.Results:1.Under the light microscope,the muscle fibers were disordered,stained unevenly,even ruptured,the nuclear distribution was disordered,and a large number of inflammatory cells appeared in the model group.Under electron microscopy,myofilolysis,degeneration,extensive mitochondrial edema and vacuolation,and a large number of autophagy and apoptotic bodies were observed in the model group.After acuputomy and EA intervention,the whole condition of rabbit neck muscle was improved obviously,and the acupotomy group was superior to the electro-acupuncture group.In acupotomy group,muscle fiber was arranged neatly,muscle satellite cells could be seen,and mitochondria were evenly distributed.2.Compared with control group,the expression level of PI3 K gene and protein in model group were decreased(P<0.05),so was the P-Akt protein.And PI3 K gene and protein in acupotomy group were apparently increased than that in model and electro-acupuncture groups(P<0.01).Compared with model group,the expression level of P-Akt protein in both acupotomy and electro-acupuncture groups were obvious increased(P<0.05).There was no statistical difference in the expression of Akt protein among each groups(P>0.05).Conclusions:1.Acupotomy therapy can improve the histopathology and ultrastructural changes of the injured neck muscles and promote the regeneration and repair of the damaged neck muscles;2.Acupotomy has a positive effect on the treatment of CS rabbits,which may be related to it can regulate the autophagy and apoptosis balance in cervical muscle cells through PI3K/Akt signaling pathway and may be a potential target for repairing cervical muscle strain.