| Objective: The adjuvant arthritis(AA)rats induced by complete Freund’s adjuvant were used as rheumatoid arthritis(RA)model to observe the changes of inflammation-related factors IL-1,TNF-α,autophagosome and autophagy-related molecules ULK1,Beclin1 and LC3 in rats induced by electroacupuncture at "Zusanli" and "Guanyuan" points,so as to explore the related effect and mechanism of electroacupuncture on rheumatoid arthritis.Methods: 24 SD rats were randomly divided into normal group(n =6),model group(n=6),methotrexate group(n=6)and electroacupuncture group(n=6).The model was made by subcutaneous injection of complete Freund’s adjuvant into the foot pad of rats.After successful modeling,the normal group and model group did not receive any treatment,the methotrexate group was given intragastric administration of methotrexate according to the dose of 0.35mg/ ㎏,and the electroacupuncture group was given electroacupuncture at Zusanli and Guanyuan points.The body mass and paw volume of rats were measured every 3 days.After treatment,the pathomorphology of ankle synovium of rats in each group was examined,the contents of IL-1 and TNF-α in serum of rats in each group were detected by Elisa method,the contents of ULK1,Beclin1 and LC3 in synovium of ankle joint were detected by Western-Blotting,and the autophagy bodies of rats in each group were observed under transmission electron microscope.Results:1.Paw volume: on the 3rd day after modeling,compared with the normal group,the paws of rats in the model group,methotrexate group and electroacupuncture group showed varying degrees of swelling,and the paw volume of rats increased significantly(P<0.01),suggesting that the model was successful.On the 10 th day after modeling,the foot volume of the model group,methotrexate group and electroacupuncture group was still significantly higher than that of the normal group(P<0.01).Compared with the model group,the foot volume of methotrexate group and electroacupuncture group decreased,but there was no statistical significance(P>0.05).On the 17 th day after modeling,the paw swelling in the methotrexate group was significantly lower than that in the model group(P<0.05).The paw swelling in the electroacupuncture group was significantly reduced(P<0.01),but there was no significant difference between the methotrexate group and the electroacupuncture group.On the 24 th day after modeling,the toe swelling in the methotrexate group and electroacupuncture group was significantly less than that in the model group(P<0.01),but there was no significant difference between the two groups.2.The results of pathological examination: the structure of synovium in the normal group was clear and the shape was normal;the synovial epithelium in the model group was not smooth,the inflammatory cell infiltration was severe,and the synovial epithelium in the methotrexate group was not smooth,part of the synovium was thickened and a small amount of inflammatory cells infiltrated;in the electroacupuncture group,the synovial epithelium was not smooth,the structure was clear,and the inflammatory infiltration was less than that in the model group.3.Comparison of the contents of IL-1 and TNF-α in the serum of rats: compared with the normal group,the contents of IL-1 and TNF-α in the serum of the model group were significantly higher than those of the normal group(P<0.01),suggesting that the level of inflammation in the serum of rats increased significantly after the establishment of the model.Compared with the model group,the contents of serum IL-1 and TNF-α in the methotrexate group decreased significantly(P<0.01),and the contents of IL-1 and TNF-α in the electroacupuncture group decreased significantly(P<0.01).Compared with methotrexate group,there was no significant difference in serum IL-1 and TNF-α contents between electroacupuncture group and methotrexate group.4.Comparison of ULK1,Beclin1 and LC3 protein expression in synovium of rat ankle joint: compared with the normal group,the expression of ULK1 protein and Beclin1 protein in synovium of model group increased significantly(P<0.05),suggesting that autophagy-related pathway may be activated after modeling,resulting in abnormal activity of autophagosomes.Compared with the model group,the expression of ULK1 protein in synovium of rats in methotrexate group decreased significantly,while the expression of ULK1 and Beclin1 protein decreased significantly in electroacupuncture group.Compared with methotrexate group,the expression of ULK1 in electroacupuncture group was significantly lower than that in methotrexate group,but there was no significant difference in Beclin1 protein expression.Compared with the normal group,the expression of LC3 protein in the model group increased significantly,and the difference was statistically significant.Compared with the model group,the expression of LC3 protein in the methotrexate group was significantly decreased,and the expression of LC3 protein in the electroacupuncture group was significantly lower than that in the model group(P<0.01).Compared with methotrexate group,there was no significant difference in LC3 protein expression between electroacupuncture group and methotrexate group.Conclusion:1.El ectroacupuncture at "Zusanli" and "Guanyuan" points can significantly improve the toe swelling of AA rats,down-regulate the expression of IL-1 and TNF-α,regulate inflammatory reaction and reduce synovial injury.2.Electroacupuncture at "Zusanli" and "Guanyuan" can regulate the autophagy of synoviocytes in AA rats,down-regulate the expression of ULK1 and LC3 in synovium of AA rats,alleviate abnormal synovial proliferation,reduce synovial injury and delay the progression of RA. |
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