| Objective: This experiment by warm acupuncture and moxibustion intervention in spinal cord injury(SCI)in rats "baihui ","dazhui" and "yaoyangguan ","zusanli" hole,temperature contrast observation acupuncture acupoints in head of SCI rats behavior and VCAM-1,Wnt-3a andβ-Catenin expression,the effect of acupuncture in the treatment of SCI rats in order to investigate temperature possible mechanism of action,to xingshentongdu warm acupuncture and moxibustion treatment of spinal cord injury to provide experimental basis and theoretical foundation.Methods: The design method of this rat experiment is random grouping,parallel control and double blind method.To prepare SCI rat model,20 rats were randomly divided into model control group,jiaji acupoint acute stage group,jiaji acupoint recovery stage group,head acupoint acute stage group and head acupoint recovery stage group,4 rats in each group,and 4 rats in normal control group.BBB scores were obtained on the 1st and 14 th day after the operation.According to the experimental design,each group of rats were given the corresponding treatment plan.Serum,spinal cord and telencephalon specimens of 24 rats were collected on the 14 th day after operation.The expression of VCAM-1 in rat serum was detected by ELISA.Real-time quantitative PCR was used to detect the m RNA expression changes of VCAM-1 and wnt3 a in spinal cord and brain of rats.Western blotwas used to observe the expression changes of wnt3 a and β-catenin proteins in spinal cord and terminal brain of rats.The differences of BBB score,VCAM-1 expression and protein expression in Wnt/β-catenin signaling pathway were compared in different groups of rats,and the relevant experimental data were statistically analyzed and processed.Results: 1.Behavioral indexes of rats in each group were observed 1 day and 14 days after the operation,the rats in each group were given BBB scores,and the comparison results were as follows: ⑴BBB score of rats in each model building group was 0 after 1 day of model building operation,no voluntary movement of the hind limbs was observed in each group,and complete paraplegia was observed in each group,indicating that the spinal cord injury model was successfully established.⑵Fourteen days after the operation,compared with the model control group,the BBB score of jiaji acupoint acute stage group,head acupoint acute stage group and head acupoint recovery group were significantly improved(P<0.05),while the BBB score of jiaji acupoint recovery group was slightly higher than that of the model control group,but there was no significant improvement(P>0.05).⑶Fourteen days after the operation,BBB score of the acupoint acupoint group was significantly improved compared with that of the jiaji acupoint group(P<0.05).The BBB score of the recovery period of acupoints in the head group was slightly higher than that of the recovery period of jiaji points,but there was no significant improvement(P>0.05).⑷Fourteen days after the surgery,BBB score in the acute phase of jiaji acupoint group was slightly higher than that in the recovery phase of jiaji acupoint group,but there was no significant improvement(P>0.05).The BBB score of acupoint acute group was higher than that of acupoint recovery group,which was statistically significant(P<0.05).2.Comparison of serum VCAM-1 content in each group The experimental results showed that the contents of serum VCAM-1 of the normal control group,the model control group,the jiaji acupoint acute stage group,the jiaji acupoint recovery stage group,the head acupoint acute stage group and the head acupoint recovery stage group were not statistically significant(P>0.05).3.Comparison of m RNA levels of VCAM-1 in spinal cord and terminal brain of rats in each group 3.1 comparison and analysis of the m RNA content of VCAM-1 in the spinal cord of each group were as follows: ⑴Compared with the normal control group,the m RNA of VCAM-1 was significantly increased in the remaining five groups(P<0.05).⑵Compared with the model control group,there was no significant increase in VCAM-1 m RNA in the jiaji acupoint acute stage group and jiaji acupoint recovery stage group(P>0.05),while there was a very significant increase in VCAM-1 m RNA in the acupoint acute group and the recovery stage group(P<0.01).⑶Compared with the jiaji acupoint acute phase group,the m RNA of VCAM-1 in the head acupoint acute phase group increased significantly(P<0.01).Compared with the convalescence group of jiaji acupoints,the m RNA of VCAM-1 in the convalescence group of head acupoints increased significantly(P<0.01).⑷There was no significant increase of VCAM-1 m RNA in the acute phase group(P>0.05).There was no significant increase of VCAM-1 m RNA in the acute acupoint group(P>0.05).3.2 the comparison and analysis of the m RNA content of VCAM-1 in the brain of each group were as follows: ⑴Compared with the normal control group,there was no significant increase of VCAM-1 m RNA in the model control group,jiaji acupoint acute phase group and jiaji acupoint recovery phase group(P>0.05),while therewas a very significant increase of VCAM-1 m RNA in the acute acupoint acute phase group and jiaji acupoint recovery phase group(P<0.01).⑵Compared with the model control group,there was no significant increase in VCAM-1 m RNA in the jiaji acupoint acute stage group and jiaji acupoint recovery stage group(P>0.05),while there was a very significant increase in VCAM-1 m RNA in the acupoint acute group and the recovery stage group(P<0.01).⑶Compared with the jiaji acupoint acute phase group,the m RNA of VCAM-1 in the head acupoint acute phase group increased significantly(P<0.01).Compared with the convalescence group of jiaji acupoints,the m RNA of VCAM-1 in the convalescence group of head acupoints increased significantly(P<0.01).⑷There was no significant increase of VCAM-1 m RNA in the acute phase group(P>0.05).There was no significant increase of VCAM-1m RNA in the acute acupoint acupoint group(P>0.05).4.Comparison of m RNA levels of wnt-3a in spinal cord and terminal brain of rats in each group 4.1 the m RNA levels of wnt-3a in the spinal cord of each group were compared and analyzed as follows: ⑴Compared with the normal control group,the m RNA of wnt-3a in the model control group,the acute phase of head acupoints group and the recovery phase of head acupoints group all increased significantly(P<0.01),while the m RNA of wnt-3a in the acute phase of jiaji point group and the recovery phase of jiaji point group did not increase significantly(P>0.05).⑵Compared with the model control group,there was no significant increase of wnt-3a m RNA in the jiaji acupoint acute stage group and jiaji acupoint recovery stage group(P>0.05),while there was a very significant increase of wnt-3a m RNA in the acute acupoint acute stage group and the recovery stage acupoint head acupoint group(P<0.01).⑶Compared with the jiaji acupoint acute phase group,the m RNA of wnt-3a in the head acupoint acute phase group increased significantly(P<0.01).Compared with the recovery group of jiaji acupoints,the m RNA of wnt-3a in the recovery group of head acupoints increased significantly(P<0.01).⑷There was no significant increase in the m RNA of wnt-3a in the acute phase of jiaji acupoint group compared with that in the recovery phase of jiaji acupoint group(P>0.05).There was no significant increase of m RNA of wnt-3a in the acute acupoint group(P=0.05).4.2 the comparison and analysis of the m RNA content of wnt-3a in the brain of each group were as follows: ⑴Compared with the normal control group,there was no significant increase of wnt-3a m RNA in the model control group,the jiaji acupoint acute phase group and the jiaji acupoint recovery phase group(P>0.05),while there was an extremely significant increase of wnt-3a m RNA in the acute acupoint acute phase group and the head acupoint recovery phase group(P<0.01).⑵Compared with the model control group,there was no significant increase of wnt-3a m RNA in the jiaji acupoint acute stage group and jiaji acupoint recovery stage group(P>0.05),while there was a very significant increase of wnt-3a m RNA in the acute acupoint acute stage group and the recovery stage acupoint head acupoint group(P<0.01).⑶Compared with the acute phase of jiaji acupoint group,the m RNA of wnt-3a in the acute phase of head acupoint group increased significantly(P<0.01).Compared with the recovery group of jiaji acupoints,the m RNA of wnt-3a in the recovery group of head acupoints increased significantly(P<0.01).⑷There was no significant increase in the m RNA of wnt-3a in the acute phase of jiaji acupoint group compared with that in the recovery phase of jiaji acupoint group(P>0.05).The m RNA of wnt-3a in the acute acupoint group increased significantly(P<0.01).5.Comparison of wnt-3a protein content in spinal cord and terminal brain of rats in each group 5.1 the comparative analysis of wnt-3a protein content in spinal cord of each group was as follows: ⑴Compared with the normal control group,the wnt-3a protein in the remaining five groups increased significantly(P<0.01).⑵Compared with the model control group,there was no significant increase in wnt-3a protein in the acute phase of jiaji acupoint group,recovery phase of jiaji acupoint group and recovery phase of head acupoint group(P>0.05),while there was a very significant increase in wnt-3a protein in the acute phase of head acupoint group(P<0.01).⑶Compared with the acute phase of jiaji acupoint group,wnt-3a protein in the acute phase of head acupoint group increased significantly(P<0.01).There was no significant increase in wnt-3a protein in the head acupoint recovery group(P>0.05).⑷Compared with the jiaji acupoint convalescence group,wnt-3a protein in the acute acupoint group increased significantly(P<0.05).Wnt-3a protein in the acute acupoint group increased significantly(P<0.01).5.2 comparative analysis of wnt-3a protein content in the terminal brain of each group was as follows: ⑴Compared with the normal control group,there was no significant increase in wnt-3a protein in the model control group,the jiaji acupoint acute stage group and the jiaji acupoint recovery stage group(P>0.05),while there was an extremely significant increase in wnt-3a protein in the acupoint acute stage group and the jiaji acupoint recovery stage group(P<0.01). ⑵Compared with the model control group,there was no significant increase in wnt-3a protein in the jiaji acupoint acute stage group and jiaji acupoint recovery stage group(P>0.05),while there was a significant increase in wnt-3a protein in the acupoint acute stage group and acupoint recovery stage group(P<0.05).⑶Compared with the acute phase of jiaji acupoint group,the wnt-3a protein in the acute phase of head acupoint group increased significantly(P<0.01).There was no significant increase in wnt-3a protein in the head acupoint recovery group(P>0.05).⑷Compared with the jiaji acupoint convalescence group,there was no significant increase in wnt-3a protein in the acute acupoint group(P>0.05).Compared with the recovery group of head acupoints,the wnt-3a protein in the acute group of head acupoints increased significantly(P<0.05).6.Comparison ofβ-Catenin protein content in spinal cord and terminal brain of rats in each group 6.1 comparison and analysis of the content of β-Catenin protein in the terminal brain of each group were as follows: ⑴compared with the normal control group,the remaining five groups of rats all showed significant increase inβ-Catenin protein(P<0.05).⑵Compared with the model control group,there was no significant increase in the extract of β-Catenin protein in the acute phase of jiaji acupoint group,the recovery phase of jiaji acupoint group and the recovery phase of head acupoint group(P>0.05),while there was a very significant increase in the extract of β-Catenin protein in the acute phase of head acupoint group(P<0.01).⑶Compared with the acute phase of jiaji acupoint group,there was an extremely significant increase in the amount of β-Catenin protein in the acute phase of head acupoint group(P<0.01).Compared with the recovery group of jiaji acupoints,there was no significant increase in the recovery group(P>0.05).⑷Compared with the jiaji acupoint recovery group,there was no significant increase of β-catenin protein in the jiaji acupoint acute phase group(P>0.05).Compared with the recovery group of head acupoints,the acute group of head acupoints showed an extremely significant increase in the amount of β-catenin protein(P<0.01).6.2 comparison and analysis of the contents of β-Catenin protein in the end brain of rats in each group are as follows: ⑴Compared with the normal control group,there was no significant increase in the extracellularβ-Catenin protein in the model control group,In addition,the jiaji acupoint acute phase group and the jiaji acupoint recovery phase group(P>0.05),while there was an extremely significant increase in the extracellularβ-Catenin protein in the acute acupoint acute phase group and the head acupoint recovery phase group(P<0.01).⑵Compared with the model control group,there was no significant increase in the extract ofβ-Catenin protein in the acute phase of jiaji acupoint group and the recovery phase of jiaji acupoint group(P>0.05),while there was a very significant increase in the extract ofβ-Catenin protein in the acute phase of head acupoint group and the recovery phase of head acupoint group(P<0.01).⑶Compared with the acute phase of jiaji acupoint group,there was an extremely significant increase in the amount ofβ-Catenin protein in the acute phase of head acupoint group(P<0.01).Compared with the convalescence group of jiaji acupoints,there was a significant increase in the amount ofβ-Catenin protein in the convalescence group of head acupoints(P<0.05).⑷The results showed that there was no significant increase ofβ-Catenin protein in the acute phase group(P>0.05).Compared with the recovery group of head acupoints,the acute group of head acupoints showed an extremely significant increase in the amount ofβ-Catenin protein(P<0.01).Conclusion: 1.The head acupoints of warm acupuncture and moxibustion had a certain improvement effect on the behavioral indicators of SCI rats,which could improve the motor dysfunction of both hind limbs in rats,and the intervention in the acute phase had a better improvement effect on the motor dysfunction of hind limbs.2.The expression of VCAM-1 in the spinal cord and the terminal brain can be significantly up-regulated by warm acupuncture and the head acupoints,and the acute phase intervention is more significant.3.The expression of key proteins Wnt-3a and β-Catenin in the Wnt/β-Catenin signaling pathway in the spinal cord and terminal brain of rats was significantly up-regulated by acupoints on the head of warm acupuncture and moxibustion,and the promoting effect of Wnt-3a andβ-Catenin in the acute phase was more significant.4.According to the legislation of awakening the mind,the acute phase intervention of SCI rats can better improve the motor dysfunction of both hind limbs,and the mechanism may be related to the obvious up-regulation of VCAM-1 and key protein expression in Wnt/β-Catenin signaling pathway by acupoints on the head stimulated by acute phase warm acupuncture and moxibustion. |
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