| Objective:Observed articular cavity injection of ozone(O3) curative effect of interference in the CIA model rats, and explore ozone in serum levels of articular cavity after injection, factors involved in bone metabolism in CIA rats serum RANKL and OPG and inflammatory chemokines, RANTES MCP- 1, the influence of CXCL10 level,further in the organization and the molecular level study of intra-articular injection of ozone RANKL/OPG of CIA rats bone destruction pathway and its important transfer protein TRAF6 and the influence of CXCL10 / CXCR3 chemokines axis.Methods:40 weight and age of rat of Wistar rats were randomly divided into 2 groups, blank control group(8) and module(32), build module with bovine type II collagen and freund’s complete adjuvant emulsion two immune animals, CIA rats model was established, after building success(24) were randomly divided into 8 O3 group, only the CIA team;MTX group 8.O3 set inside the double ankle injection concentration of40 mu g/ml of O3 each 1 ml, 1 times a week, a total of injection for three weeks.By intraperitoneal injection of MTX group 0.9 mg/kg, 1 times a week, a total of three weeks.Measured foot swelling degree, and radiographic assessment of arthritis index score curative effect evaluation of CIA rats.Detection technology with high flux flow more factor determination of each group rats serum RANKL and OPG neutrons, MCP- 1, RANTES, the level of CXCL10.Application of immunohistochemical technique testing TRAF6 synovial tissue expression and CXCR3 level;Rt-pcr method is applied to detect the lymph nodes, synovial, spleen tissues RANKL and OPG, TRAF6 and CXCL10, CXCR3 m RNA expression level;Using t test or Wilcoxon rank of each application and test compared between groups.Results:1. The articular cavity injection of ozone intervention effect on CIA rats:After 40 ug/ml of articular cavity injection of ozone foot swelling degree, and radiographic arthritis index score improved significantly, prompt articular cavity injection of ozone has definite curative effect on CIA rats2. The effects on bone destruction system:1) in serum RANKL and OPG and RANKL/OPG changes:?RANKL/OPG and RANKL: CIA group increased significantly compared with normal group, the difference was statistically significant(P < 0.05), compared with the CIA group O3 group was obviously lower, the difference was statistically significant(P < 0.05), compared with MTX group level, there was no statistically significant difference(P > 0.05).A longitudinal OPG levels: there was no statistically significant difference between the groups(P > 0.05).2) TRAF6 protein expressed in the synovium The CIA synovial cell TRAF6 protein expression was stronger than normal group,MTX group and O3 TRAF6 weak protein expression in the group with the CIA;O3TRAF6 protein expression weaker than MTX group;3) lymph nodes, synovial membrane, RANKL/OPG and TRAF6 m RNA expression in the spleen(1)lymph nodes and synovial: RANKL/OPG ratio and TRAF6: CIA group m RNA expression significantly higher than normal group, the difference was statistically significant(P < 0.05);O3 group significantly decreased in the CIA group, the difference was statistically significant(P < 0.05);There was no statistically significant difference O3 and MTX group(P > 0.05).(2) spleen: CIA group TRAF6 m RNA expression significantly higher than normal group, the difference was statistically significant(P < 0.05);O3 group significantly decreased in the CIA group, the difference was statistically significant(P <0.05);There was no statistically significant difference O3 and MTX group(P >0.05).RANKL/OPG ratio has no obvious difference between groups(P > 0.05).3. The effects of chemokines1) in serum, RANTES, CXCL10 MCP- 1 level(1) RANTES MCP- 1: CIA group increased significantly compared with normal group, the difference was statistically significant(P < 0.05), the difference of O3 and CIA group has no statistical significance(P > 0.05).(2) CXCL10: CIA group increased significantly compared with normal group, the difference was statistically significant(P < 0.05), O3 group significantly decreased compared with the CIA group, the difference was statistically significant(P < 0.05),compared with MTX group level, there was no statistically significant difference(P >0.05)2) CXCR3 in synovial membrane protein expression The CIA synovial cell CXCR3 protein expression was stronger than normal group,MTX group and O3 CXCR3 weak protein expression in the group with the CIA;O3CXCR3 protein expression weaker than MTX group3) CXCL10, CXCR3 m RNA expression in organization(1) synovial: CIA group increased significantly compared with normal group, the difference was statistically significant(P < 0.05), O3 group is significantly lower with the CIA group, the difference was statistically significant(P < 0.05).There was no statistically significant difference O3 and MTX group(P > 0.05)(2) lymph nodes and spleen O3 group with the CIA group had no statistical difference(P > 0.05)... |
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