Madecassoside Impedes Invasion Of Rheumatoid Fibroblast-like Synoviocyte From Adjuvant Arthritis Rats Via Inhibition Of NF-?B-mediated Matrix Metalloproteinase-13 Expression

Part one The expression of MMPs in RA synovial tissue and RA-FLSObjectives:To observe the expression of metal matrix protease family(MMPs)in cervical rheumatoid arthritis fibroblast(RA-FLS).Methods:Extracted from the cervical spine in patients with RA synovial tissue and cultured RA-FLS,using immunohistochemistry(IHC)and hematoxylin eosin stain(HE)staining,protein imprinting testing(WB),immunofluorescence(IF)detection method for determination of MMPs expression in cervical RA into fibrous tissue.Results:MC can relieve the symptoms of cervical RA rat,neck at the back of the injection of the MC of the treatment group rats significantly reduce joint swelling degree and the atlanto-axial joint diameter and arthritis inflammatory score has dropped significantly.And treatment of pathological changes of rats get improved significantly,cartilage injury,synovial hyperplasia,reduce inflammatory cells infiltration.Conclusion:These results indicated that MMPs were expressed in the nucleus and cytoplasm of the cervical RA-FLS,and suggested that MMPs were involved in the proliferation of RA synovial membrane in the cervical spine through NF-?B pathway.Part two Experimental study on the bone destruction of the adjuvant cervical vertebrae RA rats by hydroxyl.Objectives:To observe the migration and invasion ability of the synovial fibroblasts in cervical rheumatoid arthritis induced by MMPS.Methods:Three groups which were normal blank control group,cervical spine RA disease model group,and hydroxyl acyclovir(MC)treatment group.And purification of the MC solution,X-ray situation of rats by observing the general situation and the inflammatory swelling of the score to determine the role of cervical RA FLS-MC solution,and use the Hoechst staining and double antibody sandwich enzyme-linked immunosorbent(ELISA)to determine the MC on regulate MMPs to curb RA cervical spine bone damage in rats.Results:MC can relieve the symptoms of RA rats in the cervical spine,and the degree of joint swelling of the rats in the treatment group with MC at the back of the neck was significantly reduced,and the diameter of atlantoaxial joints and inflammatory scores of arthritis decreased significantly.In addition,the pathological changes of the treated rats were significantly improved,cartilage lesion,synovial hyperplasia was relieved,and inflammatory cell infiltration decreased.Conclusion:These results indicate that MC solution can inhibit the MMPs induced cervical RA-FLS,which can inhibit bone destruction.Part three The expression of hydroxyl glycyrrhizin via NF-?B pathway regulates the expression of mmp-13 and inhibits bone destruction in the adjuvant cervical vertebrae RA rats.Objectives:The rat model was used to study the regulation of MC through NF-?B pathway to inhibit the bone destruction of adjuvant rats by inhibiting the expression of mmp-13.Methods:The effects of MC on the binding activity of transcription factor NF-?B pathway promoter were analyzed by using the dual-luciferase reporter gene,and the expression of mmp-13 was inhibited by the down-regulation of NF-?B pathway.At the same time,by means of enzyme-linked immunoassay(ELISA),MMp-13 was used to control the secretion of inflammatory cytokines in RA of cervical vertebrae.Chromatin immunoprecipitation assay(ChIP)was used to further verify the effect of MC on NF-?B pathway and mmp-13.Results: RA-FLS cells after dealing with the MC treatment,than with the control,number of CD4 +,CD8 + T lymphocytes were large increase obviously,give play to the role of the immune conditioning section,at the same time ChIP to detect the MC for the NF-?B combined with MMP-13 promoter activity and the effect of promoting RA-FLS-cell apoptosis.In rat RA,the NF-?B was activated and the signal was expressed.After treatment by MC solution,the expression of NF-?B was significantly reduced.Like the NF-?B pathway proteins,the experimental results showed that after 24 h and 48 h after dealing with the MC,IL-2,the secretion of IL-?and IFN-?expression increased obviously,prompt the MC solution can promote the proliferation of T lymphocytes.Conclusion: MC can inhibit the expression of mmp-13 by down-regulating the NF-?B pathway,while MC can inhibit the bone destruction of RA-FLS through the above immunoregulation and cross regulation.Conclusion:1.These results indicate that MC solution can inhibit the MMPs induced cervical RA-FLS,which can inhibit bone destruction.2.MC can inhibit the expression of mmp-13 by down-regulating the NF-?B pathway,while MC can inhibit the bone destruction of RA-FLS through the above immunoregulation and cross regulation.3.These results indicated that MMPs were expressed in the nucleus and cytoplasm of the cervical RA-FLS,and suggested that MMPs were involved in the proliferation of RA synovial membrane in the cervical spine through NF-?B pathway.