| Objective:Establish animal models of cervical disc degeneration through the bow-bit+ wind cold stimulation" method,Intervention animal model with acupotomoy, From the aspect of the cervical disc tissue morphology and extracellular matrix components, metabolic enzymes and cytokines content changes to explore the acupotomoy therapy can delay or even by repairing intervertebral disc degeneration to achieve the purpose of treatment of cervical disease.Methods:Purchase 6±0.5-month-old male New Zealand rabbits and divide into four groups randomly:normal control group,model group, acupuncture group and the acupotomoy groups,N=6. except the control group, other groups were established experimental model of "bow-bit+wind cold stimulation"method. After created models,comparing the model group and the control group rabbit cervical spine X-ray to determine whether the model is successful or not.After successfully modeling,Using acupotomoy and acupuncture intervened the acupotomoy groups and acupuncture group respectively, the acupotomoy groups intervention once a week for a total of 4 times intervention, the acupuncture groups intervention 3 times a week for a total of 12 times intervention. After the intervention, HE staining, light microscope morphological changes four groups of animals neck muscles and intervertebral discs.Four groups of experimental animals to detect the disc MMP-3, TGF-β1 content by ELISA, disc II collagen, aggrecan protein levels with the Western blot.Results:Normal control animals cervical X-ray showed:normal cervical lordosis,intervertebral space without widening or narrowing, vertebral body neat, smooth and free of vertebral osteophytes around.Modeling of experimental animals after cervical spine X-ray showed:Normal cervical lordosis disappeared, straight or even anti-bow, disc space narrowing, vertebral irregular arrangement, there are rough edges vertebral osteophyte formation.After the intervention of acupotomoy and acupuncture, neck musclestaining under the light microscope showed:The muscle tissue structure of the normal control group animals is clearly visible, no muscle cell degeneration and edema, no inflammatory cell infiltration. Model group muscle fiber arrays irregularly,the nuclears of muscle cell distribut unevenly, Numerous inflammatory cells infiltrate.The muscle tissue structure of acupotomoy group is neat than the model group, inflammatory cells infiltration significantly reduce. Acupuncture group showed muscle fibers are arranged neatly compared with the model group, muscle cells still mild swelling, distortion, a small amount of interstitial inflammatory cell infiltration, but compared with the model group was significantly reduced.Disc HE staining under the light microscope showed:Normal control group nucleus centered normal size,the fiber annulus wrapped around in neat rows. Model group showed obviously condensation nucleus,fiber annulus alignment messy, distorted, and even ruptured, acupotomoy group, acupuncture group were still visible condensation nucleus, but compared with the model group, improved significantly. The fiber annulus compared with the model group was neat,small fissures.ELISA results:The MMP-3 content of the model group rabbits disc was significantly higher than the normal control group, there are great significant differences between the two (P<0.01); The acupuncture group were lower than the model group, there are significant differences (P <0.05);The acupotomoy group were lower than the model group, there are great significant differences between the two (P<0.01); The acupotomoy group were lower than the acupuncture group, there are great significant differences between the two (P<0.01); The TGF-β1 content of the model group rabbits disc was significantly lower than the normal control group, there are great significant differences between the two (P<0.01); The acupuncture group were higher than the model group, there are significant differences (P<0.05);The acupotomoy group were higher than the model group, there are great significant differences between the two (P<0.01); The acupotomoy group were higher than the acupuncture group, there are no statistically significant (P>0.05)Western blot results:The II collagen protein expression levels of four groups experimental animals, the model group was significantly lower than the normal control group, there are great significant differences between the two (P<0.01); The acupuncture group were higher than the model group, there are significant differences (P<0.05);The acupotomoy group were higher than the model group, there are significant differences between the two (P<0.05); The acupotomoy group were higher than the acupuncture group, there are great significant differences between the two (P<0.01); The acupotomoy group were higher than the acupuncture group, there are significant differences between the two (P<0.05). The aggrecan protein expression levels of four groups experimental animals, the model group was significantly lower than the normal control group, there are great significant differences between the two (P<0.01); The acupuncture group were higher than the model group, there are great significant differences (P<0.01);The acupotomoy group were higher than the model group, there are significant differences between the two (P<0.05); The acupotomoy group were higher than the acupuncture group, there are great significant differences between the two (P<0.01); The acupuncture group were higher than the acupotomoy group,there are no statistically significant (P>0.05)Conclusions:1.Compare the changes of the normal control group and model group cervical spine X-ray,neck muscles and cervical disc tissue morphological,"bow-bit+wind cold stimulation"can successfully established experimental model of disc degeneration, the experimental model was successful.2. Acupotomoy and acupuncture intervention all can make the protein expression levels of II collagen and aggrecan increased, MMP-3 content decreased, TGF-β1 content increased,indicating that acupotomoy and acupuncture intervention curing the cervical disease may through benign regulating the contents of cytokines and metabolic enzymes, inhibiting extracellular matrix degradation,to alleviate or even repair disc degeneration.3.From the comparison of neck muscle and disc tissue morphology, trace substances in the disc, Overall, the efficacy of acupotomoy is slightly better than the acupuncture. |
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