| Objective: By observing the effect of Bushen Huoxue Decoction on NLRP3/Caspase-1inflammasome in degenerated intervertebral discs of rats,this thesis expounds the mechanism of Bushen Huoxue Decoction in treating intervertebral disc degeneration,and provides molecular scientific basis for Bushen Huoxue Decoction to improve intervertebral disc degeneration.Methods: 1.50 SD rats(half male and half female)were randomly divided into three groups: 30 rats in group I were used to establish the model of intervertebral disc degeneration,10 rats in group II were fed routinely as the blank group(group A),and 10 rats in group III were used as the sham operation group(group B).After the establishment of the model,3 male and 3 female rats were randomly selected from group I,1 male and 1 female rat from group II and group III,respectively,to verify whether the model was successful.After the successful establishment of the model,the remaining 24 rats in group I were randomly divided into 8 rats in model group(group C),8 rats in inhibitor group(group D),and 8 rats in Chinese medicine group(group E),each half male and female.2.In group A,B and C,normal saline was perfused daily,in group D,inhibitor(VX-765)was perfused daily,and in group E,Bushen Huoxue Decoction was perfused daily.After two weeks and four weeks of intragastric administration,4 rats in each group were killed and L3/4～L5/6intervertebral discs were taken out.3.HE staining was used to observe the histomorphology of intervertebral discs;NLRP3 and Caspase-1 protein were extracted from tissues,and their protein contents were detected by Western blot method;IL-1β and TNF-α protein were extracted from tissues and the contents were detected by Elisa method;Total RNA was extracted from the tissue,and c DNA was obtained by reverse transcription.The relative expression of NLRP3 and Caspase-1 genes in the tissue was detected by RT-q PCR.Results: 1.HE staining results showed that two weeks after intragastric administration,the fibrous rings in group C were broken and uneven,the nucleus pulposus was shrunk,and there were fewer chordal cells in the nucleus pulposus;In group D and E,the fibrous ring showed a trend of reconstruction,the nucleus pulposus cells were relatively full,and the nucleus pulposus and chordal cells were relatively more.After four weeks of intragastric administration,the structure of fibrous ring in group C was disordered,there were fewer normal nucleus pulposus cells,more nucleus pulposus cells with no nucleus,irregular shape and vacuole shape,and angiogenesis was seen in nucleus pulposus;In group D and E,the fibrous ring gradually recovered and normal nucleus pulposus cells increased.2.Western blot results,Elisa results and RT-q PCR results all showed that no matter two weeks or four weeks after intragastric administration,in the same experimental cycle,the corresponding indexes of NLRP3,Caspase-1,IL-1β and TNF-α in intervertebral disc tissue showed that group C was significantly higher than group B(P<0.01),group D and group E were significantly lower than group C(P<0.01),and group D was significantly lower than group E(P<0.01).3.Comparisons of various indicators between the two experimental cycles in Group D,the effect of four weeks is better than that of two weeks(P<0.01).Comparisons of various indicators between the two experimental cycles in Group E,the effect of four weeks is better than that of two weeks(P<0.01).Conclusion: Bushen Huoxue Decoction can effectively inhibit the expression of NLRP3/Caspase-1 inflammasome in intervertebral discs and reduce IL-1β and TNF-α,and then,plays a role in treating the degeneration of intervertebral discs,and its effect is positively related to the medication cycle. |
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