| Objective Through the rabbit muscle injury model,to observe the prevention and treatment mechanism of Li Jin manipulation on static injury of rabbit posterior cervical muscle.Methods 63 Japanese big eared rabbits were randomly divided into three groups: blank group(21 rabbits),model group(21 rabbits),prevention and control group(21 rabbits).The three groups were divided into three dynamic observation groups(20 days,40 days,60 days).In addition to the blank group,the other rabbits were made by cervical flexion fixation.The control group was treated with tendon management until the end of 60 days observation after the model was made every day,and the model group was not treated after the model was made every day.A dynamic observation point was set up every 20 days in the whole modeling cycle to observe the prevention and treatment effect and central analgesia related indexes.1.Prevention and treatment effect:(1)local body surface temperature change: use infrared thermal imaging system to detect the neck temperature of each group of rabbits;(2)muscle histopathology: observe the pathological change of neck muscle by HE staining;(3)tissue ultrastructure: observe the ultrastructural change of muscle by transmission electron microscope;(4)muscle cytoskeleton protein: detect the expression of desmin and α-actin by immunohistochemistry.2.Central analgesic effect: enzyme-linked immunosorbent assay(ELISA)detection:(1)substance P(SP),(2)beta endorphin(β EP),(3)enkephalin(ENK),(4)cholecystokinin-8(CCK-8).The experimental data were analyzed by spss22.0.Result1.Local body surface temperature changes: in three dynamic observation points,the body surface temperature of the neck injury in the model group was significantly higher than that in the blank group(P<0.05);the body surface temperature of the neck injury in the prevention and control group was significantly lower than that in the model group(P<0.05).2.HE staining: the skeletal muscles of each group were observed by HE staining.It was found that the muscle fibers of the blank group were arranged orderly and regular in shape within 60 days.Most of the muscle fibers in the model group were disorganized,twisted andloose,accompanied by inflammatory cell infiltration.At the 20,40 and 60 day observation points,the injury degree of posterior cervical muscle gradually increased with the time axis.Compared with the model group,the injury degree of the three dynamic observation points was relatively light.3.TEM observation: the myofibrils in the blank group were arranged compactly,with clear transverse lines,alternating light and dark bands,and complete mitochondrial structure.In the model group,the myofibril damage gradually increased within 60 days.On the 20 th day,the myofibrils were arranged in disorder and the myofilaments were twisted,showing focal lesions;on the 40 th day,the myofibrils were arranged in disorder and loose,the myofilaments were dissolved and broken,a large number of vacuoles were seen,and the mitochondrial cristae were dissolved and disappeared;on the 60 th day,the myofibrils were arranged in disorder and loose,the myofilaments were twisted and broken,the sarcomeres were destroyed,the mitochondrial cristae were dissolved,and the number of mitochondria was reduced.In the 60 day observation period,the damage degree of the three observation points in the control group was less than that of the model.In general,the arrangement of myofibrils was compact and orderly,the myofilaments were rarely twisted,vacuoles were occasionally seen,the structure of mitochondria was complete,and the damage of the whole skeletal muscle fiber structure was not obvious.4.Cytoskeletal protein immunohistochemical test resultsThe average optical density(AOD)of α-actin expression in each group: comparison among groups: three observation points,the AOD value of model group was significantly lower than that of blank group(P<0.05);20 day observation point,the AOD value of control group was significantly higher than that of model group(P<0.05),and there was no significant difference between control group and blank group(P>0.05);40 day and 60 day observation point,the AOD value of control group was significantly higher than that of model group(P<0.05),and the AOD value of control group was significantly higher than that of blank group(P<0.05)Compared with the blank group,the white group was still significantly lower(P<0.05).Comparison within the group: there was no significant difference between the three observation points in the model group(P>0.05);there was no significant difference between the three observation points in the control group(P>0.05).The average optical density(AOD)of desmin expression in each group: comparison among three observation points: AOD value of model group was significantly lower than that of blank group(P<0.05);AOD value of control group was significantly higher than that of model group(P<0.05)at observation points of 20 days and 40 days,and there was no significant difference between control group and blank group(P>0.05);AOD value of control group was significantly higher than that of model group at observation points of 60 days(P<0.05),but it was still higher than that of model group(P<0.05).There was significant difference between the blank group(P<0.05).Comparison within the group: there was no significant difference between the three observation points in the model group(P>0.05);there was no significant difference between the three observation points in the control group(P>0.05).5.ELISA test resultsSP results: the SP content of cervical spinal cord and hypothalamus in the three observation points model group was significantly higher than that in the blank group(P<0.05);the SP content of the three observation points prevention group was significantly lower than that in the model group(P<0.05).The SP content of cervical spinal cord in the model group was significantly higher than that in the model group for 20 days and 60 days(P<0.05);the SP content of hypothalamus in the model group had no significant difference at three observation points(P>0.05);the SP content of thalamus in the prevention group for 60 days was significantly lower than that in the prevention group for 20 days and 40 days(P<0.05);the SP content of cervical spinal cord in the prevention group had no significant difference at three observation points(P>0.05).β-EP results: the content of β-EP in the cervical spinal cord and hypothalamus of the rabbits was significantly lower than that in the control group(P<0.05);the content of β-EP in the cervical spinal cord of the rabbits in the control group was higher than that in the model group for 40 days(P<0.05);the content of β-EP in the hypothalamus of the rabbits in the control group was higher than that in the model group for 60 days(P<0.05);the content of β-EP in the thalamus of the control group was significantly higher than that in the control group for 20 days and 40 days(P The content of β-EP in the control group was significantly higher than that in the control group on the 20 th day(P<0.05);there was no significant difference in the content of β-EP in the cervical spinal cord and hypothalamus between the three observation points in the model group(P>0.05).ENK results: the ENK content of cervical spinal cord and hypothalamus in the three observation points model group was significantly lower than that in the blank group(P<0.05);except for the 60 day observation point of hypothalamus,the ENK content of the three observation points prevention group was significantly higher than that in the model group(P<0.05).The ENK content of cervical spinal cord and hypothalamus in the control group was significantly higher than that in the control group at 20 and 40 days(P<0.05),and there was no significant difference in the ENK content of cervical spinal cord and hypothalamus in the model group at three observation points(P>0.05).CCK-8 results: the CCK-8 content of cervical spinal cord and hypothalamus in the three observation points model group was significantly higher than that in the blank group(P<0.05)(except for the 60 day observation point of hypothalamus);the CCK-8 content of cervical spinal cord and hypothalamus in the control group was significantly lower than that in the model group(P<0.05)at the 20 and 40 day observation points,and there was no significant difference between the control group and the blank group(P>0.05).The CCK-8 content in the cervical spinal cord of the model group was significantly lower than that of the model group for 40 days and 60 days(P<0.05);the CCK-8 content in the hypothalamus of the model group for 60 days was significantly lower than that of the model group for 20 days and 40 days(P<0.05);the CCK-8 content in the cervical spinal cord of the control group for 60 days was significantly higher than that of the control group for 20 days and 40 days(P<0.05);the CCK-8 content in the hypothalamus of the control group for 40 days was significantly higher than that of the control group for 20 days and 60 days(P<0.05).Conclusion1.The early prevention and treatment of Li Jin manipulation can effectively reduce the damage of pathological structure and ultrastructure of local muscle tissue,reduce the body surface temperature of the damaged part,and alleviate the damage degree of static load on skeletal muscle.2.The mechanism of the prevention and treatment of Li Jin manipulation may be related to the activation of the expression of desmin and α-actin and the promotion of cytoskeleton repair.3.The manipulation of regulating tendons can also regulate the central pain transmitters SP,β-EP,ENK and CCK-8 to achieve a certain prevention and treatment effect,and the early prevention and treatment effect is significant. |
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