| ObjectiveThis study established rabbit models of cervical spondylosis (CS) by the method of imitating human long-term flexed neck posture,applied acupotomy intervention after modeling, and compared to the group intervented by electro-acupuncture. Morphology of rabbit's cervical muscles by hematoxylin-eosin staining,and epimysium of rabbit's cervical muscles by electronic speculum were observed. All the animals were cervical X-rayed for the observation of changes of the cervical vertebrae and intervertebrale.Expression of apoptosis genes of type I collagen and TGFβ1 by RT-PCR test,and expression of apoptosis protein of type I collagen and TGFβ1 by Western blot test were conducted.Thus to observe the interaction relationship between type I collagen and TGFβ1, and it is expected to explore the relationship between fibrosis and cervical spondylosis in the neck muscle, enrich understanding of the pathogenesis of cervical spondylosis,and provide evidences for the mechanism research and efficacy of acupuncture therapy.Methods30 Six-month-old healthy adult New Zealand rabbits, equal between genders, provided by Xinglong experimental animals farms in Haidian District, Beijing,from which each weight 2.5±0.5kg.. number by weight and divided into four groups randomly:normal control group,model group, acupotomy group,electro-acupuncture(EA) group.Each group had 6 rabbits,and the rest 6 rabbits are raised to be the back-ups of model group. CS models were made by the method of imitating human long-term flexed neck posture of 45 degrees for 5 hours a day, After modeling, acupotomy and electro-acupuncture were applied to treat in acupotomy group and electro-acupuncture group. Once a week in acupotomy group, three times a week in electro-acupuncture group, both groups were treated for three weeks.Morphology of rabbit's posterior cervical muscles by hematoxylin-eosin staining,and epimysium by electronic speculum were observed.All the animals were cervical X-rayed for the observation of changes of the cervical vertebrae and intervertebrale, Expression of apoptosis genes of type I collagen and TGFβ1 by RT-PCR test,and expression of apoptosis protein of type I collagen and TGFβ1 by Western blot test were conducted.Results1.In this study, CS models were made by the method of fixing the rabbits'neck posture in specialized boxes thus simulated the pathological features of the initial status of cervical spondylosis. With the observation of morphology of cervical muscles and manifestations of cervical X-ray, the results verified model of CS was established successfully and the modeling method is reliable.And acupotomy therapy has good effects in regulating the local inflammatory response, and promoting damaged tissue repairation,and the effect of the acupotomy therapy is better than EA.2.The results of Real-time PCR tests showed that,in comparison with normal control group after modeling,expression of apoptosis genes of type I collagen increased significantly in other 3 groups, there was a highly significant difference between the model group and normal control group (P<0.01);also with the EA group and normal control group (P<0.01); and significant difference between the model group and acupotomy group (P<0.05); also significant difference between the acupotomy group and EA group (P<0.05).3. The results of Western blot tests showed that, in comparison with normal control group after modeling, expression of apoptosis protein of type I collagen increased significantly in other 3 groups, and there was a highly significant difference between the model group and normal control group (P<0.01);also with the EA group and normal control group (P<0.01);and significant difference between the model group and acupotomy group (P<0.05);also significant difference between the acupotomy group and EA group (P<0.05).4. The results of Real-time PCR tests showed that, in comparison with normal control group after modeling, expression of apoptosis genes of TGF-β1 increased significantly in other 3 groups, there was a significant difference between the model group and normal control group (P<0.05);also with the acupotomy group and normal control group (P<0.05);the acupotomy group and EA group (P<0.05).;and highly significant difference between the EA group and normal control group (P<0.01).5. The results of Western blot tests showed that, in comparison with normal control group after modeling, expression of apoptosis protein of TGF-β1 increased significantly in other 3 groups,and there was a highly significant difference between the model group and normal control group (P<0.01);also with the EA group and normal control group (P<0.01);and significant difference between the model group and acupotomy group (P<0.05);also significant difference between the acupotomy group and EA group (P<0.05).ConclusionIn this experiment, the X-ray observations can not only display lesion location, extent and degree of physiological bending, vertebral hyperostosis,intervertebral space changes the image changes from a flat film, C3-C6 between the making model was successful and reliable basis. Taking into account the modeling time of the cervical syndrome, cervical spondylosis in the 3 months of the factors in this experiment as a preliminary stage of cervical spondylosis. Provided a favorable basis.From the local organization of HE staining and electron microscopy results showed that acupuncture therapy can effectively reduce the collagen weakness of the lesion scar tissue formation, needle knife method can improve the back of the neck muscle fibrosis. Acupuncture therapy, the muscle fibers arranged in recovery neat, indicating that acupuncture therapy can not only effectively prevent pathological scar formation, and the local static and dynamic equilibrium can be restored, so that the organization does not affect muscle function after recovery. Directly with the needle knife scar local tissue shovel aside, cutting related.Three experimental observation of the needle knife and electric rabbit back of the neck muscle of type I collagen in the back of the neck muscles in the rabbit model RT-PCR analysis showed that the level of type I collagen gene expression results showed that after modeling, model group and blank group between the level of type I collagen gene expression increased significantly there are very significant differences (P<0.01)between the knife group with the model group, the level of type I collagen gene expression levels have decreased significantly and there are significant differences (P<0.05) with a blank between groups was no significant difference (P> 0.05), the knife group detected close to the blank group, indicating that acupuncture therapy in cervical spondylosis rabbit model of type I collagen gene level expression of good regulation.Western-Blot shows in the back of the neck muscles in the rabbit model of type I collagen protein levels increased significantly, compared to the model group and blank group had significantly (P<0.01), indicating a rabbit model of cervical disease through fibrosis The main factors of type I collagen cause muscle fibrosis. Type I collagen levels between the knife group with the model group expression levels have significantly reduced there are significant differences (P<0.05),the knife group detected close to the blank group, indicating that acupuncture therapy in cervical spondylosis rabbit model of type I the level of collagen expression of good regulation.In the back of the neck muscles in the rabbit model RT-PCR analysis showed that the level of cervical spondylosis rabbit model of TGF-β1 gene expression was significantly increased after modeling, model group and blank group was highly significant (P<0.01). illustrates the rabbit model of cervical disease and fibrosis of the signal of the substance released by TGF-β1 gene. The expression levels of TGF-β1 gene level compared with the model in acupuncture therapy significantly reduced, significant changes (P<0.05)between the knife group and model group, closer to the blank control group, showed good regulation. Significant difference (P<0.05) between the knife group and EA group. Show acupotomology better than the EA differences.From the Western-Blot back of the neck muscles in the rabbit model of TGF-β1 protein level was significantly increased compared to the model group and blank group there was a significant difference(P<0.01) a rabbit model of cervical spondylosis caused by the local long-term strain so that the body in order to be able to adapt the strain increase of TGF-β1 protein expression of TGF-β1 is the most rapid collagen synthesis stimulating factor to promote the process of muscle fibrosis.Changes between the knife group and model group were significantly (P<0.05) and the knife group and EA group, significant differences (P<0.05), indicating the knife treatment of cervical spondylosis rabbit model of TGF-β1 good regulation, as well as close to normal levels, as well as acupuncture therapy strengths and electro-acupuncture treatment.
|
PDF Full Text Request