| Objective: to study the epithelial mesenchymal transition phenomenon(epitheliamesenehymalatrnsiton,EMT)in the synovial tissue of rheumatoid arthritis from the perspective of tumor pathogenesis,with the aim of clarifying the role of EMT phenomenon in the development of RA disease,and to explore the intervention mechanism of Wen Yang Tongluo Formula in this way.Methods: 105 DBA/1 mice were replicated as a stable CIA animal model and divided into blank group,model group,Wenyang Tongluo Formula group and ETN group by random number table method;the inflammation of mice joints was observed by AI score,the ankle joints were scanned and imaged by microCT,and the cortical bone and bone trabeculae were quantified.The synovial tissues were collected and the differences in the expression of the corresponding proteins(E-cad,VM and FN)in the synovial membranes of mice and the expression of bone transformation-related indicators(MMP9,BMP7,OPG)between the experimental groups were examined by immunohistochemistry and western blot.Results :1.AI scores of mice: After modeling,compared with the blank group,the AI scores of mice in the model group,the ETN group and the Wenyang Tongluo Fang group increased significantly,and the difference was statistically significant(P<0.05);after 28 d of treatment,compared with the model group,the AI scores of mice in the ETN group and the Wenyang Tongluo Fang group decreased significantly(P<0.05),and the AI scores of mice in the Wenyang Tongluo Fang group were higher than those in the ETN group.2.Micro-CT:After moulding,compared with the blank group,the ankle joint of the experimental mice in the model group was obviously swollen,the bone of the ankle joint was obviously eroded and severely damaged,the bone surface of the joint was rough and uneven in all parts,and the integrity of the joint was obviously damaged;the swelling of the joint of the mice in the model group in the ETN group and Wenyang Tongluo Fang group was reduced,the pits on the bone surface were improved,and the expression of the bone parameters BMD,BV/TV and Tb.N were There was an increase in the expression of bone parameters BMD,BV/TV and Tb.N.The differences were statistically significant(P<0.05),but there were no significant differences in the expression of TV,BV and Tb.th in the other groups.Immunohistochemistry: compared with the blank group,the expression of E-cad protein in the synovial tissues of mice in the model group was significantly increased,and the difference was statistically significant(P<0.05).4.Immunoblot: compared with the blank group,the expression of E-cad protein in the synovial tissues of mice in the model group was significantly decreased.The expression of E-cad was significantly lower in the model group compared with the blank group(P<0.05);the expression of VM,FN,MMP-9,BMP-7,OPG and other proteins were significantly higher in the model group(P<0.05);compared with the model group,the above values were significantly different in the blank group,ETN group and Wenyang Tongluo Fang group(P<0.05);there was no significant difference between the Wenyang Tongluo Fang group and ETN group.Conclusion:1.Both ETN and Wenyang Tongluo Fang have the effect of treating arthritis in mice,but the efficacy of Wenyang Tongluo Fang is inferior to that of ETN.2.The three-dimensional reconstructed graphs showed that ETN and Wenyang Tongluo Fang had some osteoprotective effect,but there was no significant increase in bone volume.3.Wenyang Tongluo Fang could up-regulate E-CAD expression and down-regulate VM and FN to inhibit EMT phenomenon.4.The protective effect of Wenyang Tongluo Formula on bone destruction was related to the down-regulation of related proteins MMP-9,BMP-7 and OPG. |
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