| Purpose:The thesis to observe the histomorphological changes of nucleus pulposus and endplate of degenerative lumbar intervertebral disc in rabbits,and to detect the changes of MMP-13 protein content in nucleus pulposus and the gene expression of COL-2a.To study the therapeutic mechanism of Duhuo Jisheng Decoction on degenerative intervertebral disc at molecular level,so as to provide experimental basis for Duhuo Jisheng Decoction in treating degenerative lumbar intervertebral disc,and to make sure that Duhuo Jisheng Decoction To delay the degeneration of intervertebral disc,provide strong experimental support and basis for clinical application of Duhuo Jisheng Decoction to prevent and treat the degeneration of intervertebral disc,and promote the development of the characteristics of traditional Chinese medicine.Methodology:Eighty adult healthy New Zealand white rabbits were randomly divided into blank control group,sham operation group,model control group,traditional Chinese medicine group and high concentration Chinese medicine group,with 16 rabbits in each group.Except the blank control group and the sham operation group,the other groups established the rabbit lumbar degeneration model by fibrous annulus puncture method.After the model was successfully established,the drug dosage needed for the experimental rabbits was calculated according to the conversion formula of human and animal surface area.The blank control group,the sham operation group and the model control group were intragastrically administered with normal saline daily.The Chinese medicine group and the high concentration Chinese medicine group were given low and high concentration Duhuo Jisheng decoction,respectively.The whole course lasted for 8 weeks.All rabbits were fed normally and weighed every 1 week.The dosage of gavage was adjusted according to the change of body weight.Four rabbits in each group were sacrificed at random after 2 weeks,4 weeks,6 weeks and 8 weeks respectively.The histomorphological changes of intervertebral discs were observed by HE staining under 400-fold light microscopy;the expression of COL-2a gene in nucleus pulposus was detected by RT-PCR;the protein content of MMP-13 in nucleus pulposus was detected by Western Blot.Results:The results showed that there was no significant difference in the expression of COL-2a and MMP-13 protein between the blank group and the sham-operated group(P > 0.05);the content of COL-2a and MMP-13 protein in the intervertebral disc of the model control group was higher than that of the blank group and the sham-operated group,and the difference was statistically significant(P < 0.05),indicating that the model was successful;the content of COL-2a and MMP-13 protein in the Chinese medicine group and the high concentration Chinese medicine group were lower than that of the model control group The difference was statistically significant(P < 0.05),indicating that Duhuo Jisheng Decoction could effectively delay the degeneration of rabbit intervertebral disc;there was no significant difference in the expression of COL-2a and MMP-13 protein between the traditional Chinese medicine group and the high-concentration Chinese medicine group(P > 0.05),indicating that the concentration of traditional Chinese medicine had no significant difference in delaying the degeneration of rabbit intervertebral disc.Conclusions:1.Fibrous annulus puncture via muscle space approach can successfully create a rabbit model of intervertebral disc degeneration.2.The histological structure and pathological changes of degenerated rabbit intervertebral disc coincide with those of human degenerated intervertebral disc.Fiber ring puncture,i.e.external physical stimulation factors,can accelerate the degeneration of intervertebral disc and mimic the animal model of intervertebral disc degeneration.3.The expression of MMP-13 protein in lumbar intervertebral disc tissue was significantly decreased and the gene expression of type Ⅱ collagen was significantly increased after administration of Duhuo Jisheng Decoction to rabbits with lumbar intervertebral disc degeneration.It was proved that Duhuo Jisheng Decoction could inhibit the activity of MMP-13 protease and increase the expression of key proteins(type Ⅱ collagen)in the nucleus pulposus of intervertebral disc,thus delaying the expression of type Ⅱ collagen.The role of disc degeneration. |
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