Study On The Therapeutic Mechanism Of Yaobishu On Prolapse Of Lumbar Intervertebral Disc Based On JAK2/STAT3 Signaling Pathway

Objective: This study attempts to explore the mechanism of action of Yaobishu in the treatment of lumbar disc herniation from the perspective of JAK2 /STAT3 signaling pathway,so as to provide experimental evidence for the treatment of the disease with traditional Chinese medicine.Method:48 New Zealand white rabbits were numbered from 1 to 48 and then divided into groups by random number table method.According to 16 rabbits in each group were divided into Yaobishu treatment group,control group and blank group.The the fiber ring puncture method was carried out to carry through a lumbar disc herniation model operation in the rabbits of the Yaobishu group,and the lumbar disc herniation model operation was performed by the rabbit of the control group with the same method,and the rabbit of the blank group was not operated.The rabbits in each group were treated with drugs 8 weeks after operation:(1)Yaobishu group: 10ml/kg/ day of Yaobishu was administered by gavage;(2)Control group: 10ml/kg/ day of normal saline was administered by gavage;(3)Blank group: 10ml/kg/ day of normal saline was administered by gavage.Four animals in each of the three groups were randomly killed at the 1st,2nd,4th and 6th weeks after treatment.X-ray examination,anatomical naked eye observation and tissue section observation were performed.Simultaneously,Real-Time-PCR and Western blot methods were used to detect the level of JAK2 and STAT3 m RNA and the expression level of JAK2 and STAT3 proteins in nucleus pulposus of intervertebral disc between the three groups.SPSS18.0 software was used to analyze the statistical data of the experimental results.One-way ANOVA(F-test)was used for comparison between groups.Result:1.X-ray examination results: The Yaobishu group showed intervertebral space stenosis and hyperosteogeny changes after modeling,but gradually improved with the extension of administration time;The control group showed intervertebral space stenosis and hyperosteogeny after operation,and slowly worsened with time.The blank group showed normal lumbar X-ray films at all time periods.2.Anatomical observation results: Bone hyperplasia,fiber ring protruding and other lesions can be seen in Yaobishu group after modeling,but they gradually improve with the prolongation of administration time.Bone hyperplasia and fibrous ring protruding were observed after modeling in the control group and gradually worsened with time.Anatomical observation of blank group was normal at all stages.3.Tissue section observation results: pathological changes such as disruption of intervertebral disc tissue morphology and fibrous ring necrosis occurred in the Yaobishu group after modeling,but gradually improved with the prolongation of administration time;The control group showed disruption of intervertebral disc tissue morphology and fibrous ring necrosis after operation,and gradually worsened with time;The results in the blank group were normal in all periods.4.PCR test results: There was no significant statistical difference in the amplification factor of JAK2 and STAT3 m RNA between the Yaobishu group and the control group 1 week after treatment,but the amplification factor ratio of JAK2 and STAT3 m RNA in Yaobishu group and control group was significantly higher than that of blank group.The amplification factor of JAK2 and STAT3 m RNA in Yaobishu group gradually decreased with the prolonged administration time.The amplification factor of JAK2 and STAT3 m RNA in control group gradually and slowly increased with the administration time.The amplification factor of JAK2 and STAT3 m RNA in blank group basically at the same level as the treatment time prolonged.5.Western blot test results: There was no significant statistical differences in the expression of JAK2 and STAT3 protein between the Yaobishu group and the control group 1 week after treatment,but the test results of the Yaobishu group and the control group were significantly higher than that of blank group.The expression of JAK2 and STAT3 protein in Yaobishu group decreased gradually with the prolongation of administration time.The expression of JAK2 and STAT3 protein in the control group increased gradually with the treatment time.The expression levels of JAK2 and STAT3 proteins in the blank group remained at the same level with the prolongation of administration time.Conclusion:The occurrence and development of lumbar disc herniation is closely related to the excessive activation of the JAK2 / STAT3 signaling pathway.Yaobishu can effectively inhibit the JAK2 / STAT3 signaling pathway,reduce the level of JAK2 and STAT3 m RNA nucleus pulposus and the expression level of JAK2 and STAT3 protein,which can alleviate intervertebral disc degeneration to a certain extent.