The Effects Of Electroacupuncture On Apoptosis In Rat Model Of Chronic Cervical Muscle Injury

Objective: To observe the effect of Electroacupuncture on apoptosis and expression of bax,bcl-2,Caspase-3 and Caspase-9 proteins in chronic neck muscle injury model of rats.Methods: Twenty-eight Wistar rats of SPF grade were selected in the experiment.After a week of adaptation,the rats were divided into blank group,model group,meloxicam group and electro-acupuncture group,with an average of 7 rats in each group according to the random number table method,the other 4 were used only for the validation of successful modeling.Blank group only do the binding,and the blank group was fed with the rest of the rats as a control group.Except for the blank group,the other groups of rats were modeled by chronic neck muscle injury model,which imitated the state replication model of the state of human bowing to the desk.Rats were placed in a special fixator for cervical anterior flexion in rats.The head and neck of rats were fixed at 45 degrees of anterior flexion.The degree of tightness was based on the criterion of not affecting the normal breathing of rats.The fixing time was 5 hours,once a day,and the model was repeated for 3 months.At the end of modeling,two rats were randomly selected from the blank group and the rats after modeling.Transmission electron microscopy was used to detect the morphological changes of the posterior cervical muscle.After the modeling was completed,rats were treated with intervention.Electro-acupuncture group: Fixed rats with self-made fixed board,needled "Fengchi acupoint"(both sides),connected with Korean acupoint nerve stimulation apparatus.The stimulation parameters were as follows: the frequency of density wave was 2/100 Hz,the frequency of density wave was alternating,once a day,25 minutes per time,10 consecutive days was a course of treatment,the interval between courses was 2 days,a total of 2 courses of treatment.Meloxicam group: Meloxicam tablets were given intragastric administration once a day for the same course as electro-acupuncture group.Model group: fixed without treatment.After the intervention,the muscle tissues of rats in each group were taken,and the expression levels of bax,bcl-2,Caspase-3 and Caspase-9 proteins were detected by immunohistochemistry,and the apoptotic rate was detected by Tunel method.The other part of rat muscle tissue was used to prepare electron microscopic specimens and observe the morphological changes of muscle under electron microscopy.Results:(1)The results of electron microscopy were as follows: Blank group: It can be seen that myofibril structure is closely arranged,transverse striations are obvious,A-zone,Z-zone,I-zone,H-zone,and M-line are clear and arranged alternately and regularly,muscular serosa is intact without rupture,muscular karyotype is normal,nuclear membrane is smooth,mitochondrial structure is not abnormal and evenly distributed,and cristae are not fractured.Model group: The discontinuous arrangement of myofibril structure was slightly disordered,some mitochondria swelled and vacuoles appeared,occasionally mitochondria of different sizes were observed,and the muscle gap was almost normal.Electroacupuncture group: A small amount of myofibrils have discontinuous structure and relatively disordered arrangement,clear transverse striations,complete sarcomere and serosa,swelling of individual mitochondria,A-zone,Z-zone,I-zone,H-zone,and clear but discontinuous M-line.Meloxicam group: myofibril structure is discontinuous and cell arrangement is disordered,and muscle serosa rupture and obvious transverse striation can be seen,A-zone,H-area,Z-line,I-zone and M-line are clear but discontinuous.(2)Tunel method: The apoptotic cells of rats were brown or summer tan.Compared with the blank group,the number of apoptotic cells in the model group was more;compared with the model group,the number of apoptotic cells in the electro-acupuncture group and the meloxicam group decreased significantly.All difference have statistical significance.(P<0.05)(3)Immunohistochemical results Compared with the blank group,the positive expression of Bcl-2 in the model group was less than that in the blank group,while the positive expression of Bax,Caspase-3 and Caspase-9 increased relatively.All difference have statistical significance.(P<0.05)Compared with the model group,the positive expression of Bcl-2 in EA group and meloxicam group increased,while the positive expression of Bax,Caspase-3 and Caspase-9 decreased.(P<0.05)Conclusion :(1)Electroacupuncture can effectively inhibit cell apoptosis in rat model of chronic cervical muscle injury.(2)Electroacupuncture can down-regulate the apoptotic rate and promote the expression of Bcl-2.At the same time,it can inhibit the expression of Bax,Caspase-3 and Caspase-9 proteins,inhibit cell apoptosis,effectively promote cell proliferation and repair damaged muscle cells.