| BackgroundRheumatoid arthritis is etiologies unknow chronic inflammatory and immunological disease, characterized by an invasive synovial hyperplasia that leads to cartilage and bone destruction. The disease has a serious influence of the quality of life for its high disability. As one of Chinese Traditional Nuring Techniques, moxibustion could improve the clinical symptoms with good efficacy and relatively small adverse reactions. While the critical mechanistic of moxibustion was still remained unknown, let along the proteomic studies.ObjectivesTo elucidate the mechanism of moxibustion in treating rheumatoid arthritis by analyzing the differential expression proteins of synovitis proteomics. And provide modern scientific evidence for moxibustion in further clinic utilize.Methods(1)The CIArat model was established. The succed model CIA rats were randomly divided into six groups, the moxibusition Ⅰ group, the moxibusitionⅡgroup, the moxibusitionⅢgroup, and CIA group Ⅰ, CIA group Ⅱ, CIA groupⅢ, each of those group has six animals. The moxibustion groups select point Zhu sanli and Shen shu for treatment. No treatment were given to CIA groups. And set normal group Ⅰ, normal group Ⅱ, normal groupⅢ, also each of them has six animals.(2) Baseline comparsion:weight and the degree of paw swelling among all the groups.(3) Comparsion the general condition after three weeks moxibustion among those groups of moxibusition Ⅲ, CIA Ⅲ and normal Ⅲ.(4) Comparsion the weight and degree of paw swelling before moxibustion, a week, two weeks and three weeks after moxibustion among those groups of moxibusition Ⅲ, CIA Ⅲ and normal Ⅲ.(5) Comparsion the arthritis index scores before moxibustion, a week, two weeks and three weeks after moxibustion between the group ofmoxibusition Ⅲ and CIA Ⅲ.(6) Comparsion the HE staining after three weeks moxibustion among those groups of moxibusition Ⅲ, CIA Ⅲ and normal Ⅲ.(7) The total proteins of a week, two weeks and three weeks synovial tissue of rat in all the nine groups were separated by two-dimensional gel electrophoresis (2-DE) respectively. Choice the common proteins among the three period as the target protein for moxibustion treatment. Pep tide mass fingerprints (PMFs) obtained by the MALDI-TOF MS analysis were used to search NCBI database by using Mascot software, and the target protein then treated by the IPA analysis for biological function, signaling pathway and signaling network.Results(1) The baseline of weight and the degree of paw swelling among all the groups before modeling had no statistical significance (P>0.05)(2) After three weeks moxibustion the general condition of the normal Ⅲ group is ok,but we found a loss of apitide, lake of movement and loss of weight in the CIA Ⅲ group,and for moxibusition Ⅲ, the condition was somewhat improved.(3) The weight and degree of paw swelling of moxibusition Ⅲ and CIA Ⅲ group had a remarkable difference with the normal Ⅲ group (P<0.05), a loss of weight and welling of paws were viewed. Moxibusition Ⅲ group had a tendency of increasing the weight and decreasing the swelling, however had no difference with CIA Ⅲ group (P>0.05)(4)Before moxibustion the arthritis index scores of moxibusition Ⅲ and CIA Ⅲ group were all scored higher than four. Moxibusition Ⅲ group had a tendency of decreasing the scores, however had no difference with CIA Ⅲ group (P>0.05)(5) After three weeks moxibustion normal Ⅲ group showed no markable changes in the HE staining.However marked tissues swelling, blood vessels dilation, and accumulation of synovial fluids in the CIA rats were observed. While such pathological changes were much less in the moxibustion treated rats.(6) Well-resolved and reproducible 2-DE maps of synovial from normal, moxibustion and model groups were acquired. The average match spots of each gel were 1316.7±24.7, the average match rate was between 90% and 93%.365 proteins were found after a week moxibustion,332 proteins were found after two weeks moxibustion,202 proteins were found after three weeks moxibustion. And 9 were found among the three periods.All of the nine differential expression protein spots were identified in NCBI database. The functions of these proteins involve in Organismal Injury and Abnormalities, Skeletal and Mucular Disorders, Immunological Disease, Inflammatory Response and Disease, Hematological Disease, Connective Tissue Development and Function.et al.(7) IPA analysis found that PRDX I,EHHADH,MLC3 and HPX mainly through adjusting Fatty Acidβ-oxidation Ⅲ,NRF2-mediated Oxidative Stress Response, Acute Phase Response Signaling, Acin Cytoskeleton Signaling as well as Epithelial Adherens Junction Signaling pathways participate in the network of DNA Replication, Recombination, and Repair, Cellular Compromise and Inflammatory Response, finally involving in the onset and development of RA.Conclusions:(1) Moxibustion could improve the appetite and general coditiona of CIA rats. Less synovial cell edema as well as accumulation of inflammatory cells were viewed after three weeks moxibustion, indicated that moxibustion has anti-inflammtory effects, but the redness and swelling of the joints is difficult to relieve.(2) Nine differential expression proteins related to moxibustion were identified, indicate that synovial tissue protein of CIA could be regulated by moxibustion. The target protein PRDX I, EHHADH,MLC3 and HPX mainly through adjusting NRF2-mediated Oxidative Stress Response, Fatty Acidβ-oxidation Ⅲ, Acute Phase Response Signaling, Acin Cytoskeleton Signaling as well as Epithelial Adherens Junction Signaling pathways participate in the network of DNA Replication, Recombination, and Repair, Cellular Compromise and Inflammatory Response, finally involving in the onset and development of RA. |
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