| Objective:Rheumatoid Arthritis(RA)is a chronic inflammatory autoimmune disease with a high prevalence worldwide.A large number of studies have demonstrated the anti-inflammatory effect of moxibustion on RA,and the nuclear factor kappa B(NF-κB)signaling pathway has a significant role in the inflammation of the synovial membrane and bone destruction during RA pathology.The phosphorylation of NF-kappa-B inhibitor alpha(IκB-α)is an important factor in the activation of this pathway,and IκB kinase β(IKKβ)is the main upstream regulatory protein of IκB-α phosphorylation.Therefore,in this study,IκB-α was used as the entry point and "IKKβ/IκB-α-inflammatory factor" was used as the main line of research,and protein immunoblot(Western Blot,WB)and enzyme-linked immunosorbent assay(ELISA)were used to observe the effects of IκB-α on inflammation.The study was conducted to observe the anti-inflammatory effect of moxibustion on experimental RA rats,to investigate the mechanism of moxibustion on IKKβ/IκB-α,the key protein of NF-κB pathway,to analyze the mechanism of the effect of moxibustion on NF-κB pathway,and to further elucidate the mechanism of the anti-inflammatory effect of moxibustion on RA,so as to provide some experimental basis for clinical moxibustion treatment of RA.Methods:A total of 30 male adult SD rats(weighing 180 ±20g)were randomly divided into blank control group(blank group),RA model group(model group),moxibustion treatment group(moxibustion group),IκB-αphosphorylation inhibitor Bay11-7082 group(medicine group),moxibustion + inhibitor group,6 rats in each group.On the first day of the experiment,SD rats in the model group,the moxibustion group,the drug group and the medicine and moxibustion group were injected with 0.5 ml/kg of F?rster’s complete adjuvant into the hindfoot pads bilaterally.One week after modeling,rats in the moxibustion and drug-aid groups were treated with moxibustion of "BL23" and "ST36" with 5 strokes of each point,once/day,alternating sides.Bay 11-7082(1 mg/kg)was administered intraperitoneally to the rats in the drug group and the medicine and moxibustion group,once/day,for 6 days for both interventions,for a total of 3 sessions,with a 1-day break between sessions.The right foot pad thickness of each group of rats was measured on days 1,7 and 28,respectively;serum,left ankle synovial tissue and right ankle joint samples were collected on day 28 to observe the pathological morphology of rat synovial membrane,and the levels of serum IKKβ,IL-4and IL-17 were measured by ELISA,and the expression of IκB-α and p-IκB-α protein in synovial tissue was measured by WB method.Results:1.moxibustion can improve the foot pad thickness in RA rats.The results of the histopathological morphological examination of rat synovial tissue showed that the hyperplasia and inflammatory cell infiltration and macrophage proliferation were significantly increased in the model group compared with the blank group(P<0.05).Compared with the model group,synovial tissue hyperplasia and inflammatory cell infiltration and macrophage proliferation were decreased in the moxibustion group(P <0.05).2.The results of serum IKKβ protein assay showed that the serum IKKβ level was increased in the model group compared with the blank group(P<0.05).Compared with the model group,the serum IKKβ levels of rats in the moxibustion and drug groups decreased(P<0.05),and the IKKβ levels in the moxibustion + inhibitor groups decreased insignificantly.There was no significant difference in serum IKKβ levels between the moxibustion and drug groups,and the serum IKKβ levels in the moxibustion + inhibitor group were higher than those in the drug group(P<0.05).3.The results of synovial tissue IκB-α and p-IκB-α protein assay showed that compared with the blank group,the p-IκB-α protein level was increased in the model group rats(P<0.01),and the IκB-α level was elevated but not significantly different.Compared with the model group,the p-IκB-α protein level decreased but not significantly different in the moxibustion group rats,and the IκB-α protein level increased(P<0.05);the p-IκB-α protein level decreased(P<0.01)and the IκB-α protein level increased but not significantly different in the drug group rats;the p-IκB-α protein level decreased(P<0.01)and the IκB-αprotein levels increased(P<0.05)in the moxibustion + inhibitor group;the p-IκB-α/IκB-αratios of rats in the moxibustion,drug and moxibustion + inhibitor groups were lower than those in the model group(P<0.01).Compared with the drug group,there were no significant changes in the p-IκB-α and IκB-α protein levels in the synovial membrane of rats in the moxibustion and moxibustion + inhibitor groups.4.The results of serum cytokine IL-4 and IL-17 assay showed that compared with the blank group,the serum IL-4 level decreased but not statistically significant and IL-17 level increased in the model group rats(P<0.05).Compared with the model group,the serum IL-4 level of rats in the moxibustion group increased but not statistically significant,and IL-17 level decreased(P<0.05);the serum IL-4 level of rats in the drug group increased(P< 0.01),and IL-17 level increased(P<0.01);the serum IL-4 level of rats in the moxibustion+ inhibitor group increased(P<0.01),and IL-17 level increased(P<0.01).Compared with the drug group,the serum IL-4 level of rats in the moxibustion group was lower(P<0.01),and the IL-17 level decreased(P<0.01);the serum IL-4 and IL-17 levels of rats in the moxibustion + inhibitor group did not show significant changes.Conclusions:1.moxibustion had a significant anti-inflammatory effect on RA and inhibited the local inflammatory response in the joints of experimental RA rats.2.Moxibustion inhibits serum IKKβ levels and promotes synovial IκB-α expression in experimental RA rats,which acts on NF-κB pathway by regulating IKKβ/IκB-α expression and may directly act on subsequent inflammatory factors to achieve anti-inflammatory effects on RA. |
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