| Background:Rheumatoid arthritis is one of the chronic autoimmune diseases,which is characterized by chronic,symmetrical,and multi arthritis.According to traditional Chinese medicine(TCM)theory,RA is named as a "bi" symptom.Moxibustion is one of the TCM therapies for "bi" symptoms,with the meridians,qi and blood circulation effect.According to ancient literature,moxibustion has been applied to the treatment of "bi" symptom.Until now,animal experiments have proved that moxibustion interventions can reduce inflammatory reactions in rats and rabbits with CIA,which is an animal model of RA,through suppressing IL-1?,TNF-?.The preliminary work of our research team discovered a predicted proteomic network related to the moxibustion mechanism of CIA from the serum proteomics.Proteomic pattern analysis can identify the potential target molecules in the effect of moxibustion and understand the possible mechanism of treatment.However,comparing with proteomics,metabolomics is concerned with the final product of all metabolic processes in different biological systems.Based on these researches,the aim of this study was to discovery the biochemical changes in the serum and synovial tissue of moxibustion treatment in rats with CIA by GC-MS,which may improve the understanding the therapeutic mechanism of moxibustion.Objective:Establishing CIA rats models,and evaluating the influences of moxibustion in weight,hind paw volume,AI,related inflammatory cytokines and the pathology of ankle joints.The metabolic profiles of serum and articular synovium in CIA rats were analyzed by GC-MS.To explore the possible mechanism of its effective in the moxibustion treatment of CIA rats.Methods:(1)24 male SD(Sprague Dawley)rats were divided into the control group,the model group and the moxibustion group according to the random number table,8 rats in each group.The rats in the model group and moxibustion group were established in type? CIA.After establishing rats model successfully,rats in moxibustion group received moxibustion treatment in "Shenshu"(BL23),"Zusanli"(ST36).One treatment course consisted daily moxibustion sessions for 6 days taking one day off per course and each rat was received 3 courses of treatment.(2)Baseline comparison:body weight and hind paw volume of rats in each group before modeling.(3)The body weight:measuring the changes of body weight in 3 groups after establishing the CIA model successfully and moxibustion treatment.(4)The hind paw volume:measuring the changes of hind paw volume in 3 groups after establishing the CIA model successfully and moxibustion treatment.(5)AI scores:the scores of AI were observed after establishing the CIA model successfully and moxibustion treatment.(6)Inflammatory factors:collecting 3 groups of rats serum to compare the level of rats serum IL-1??TNF-? and PGE2 by ELISA.(7)The pathological tissue of the ankle joint of rats:the left ankle joints were collected and pathological histomorphological results were observed,after the intervention of moxibustion.(8)After the intervention of moxibustion,the serum and articular synovium tissue of 3 groups were collected.The metabolic fingerprints of serum and articular synovium tissues of 3 groups were analyzed with GC-MS technique and multivariate statistical analysis.MetaboAnalyst3.0 and related literature were used to explore the metabolic products and metabolic pathways involved.Results:(1)The baseline of weight and the degree of hind paw swelling among all the groups before modeling had no statistical significance(P>0.05).(2)Weight comparison:there was no significant difference in weight between the 3 groups(P>0.05)before establishing the CIA model,and the baseline was comparable.Compared with the control group,the weight gain in model group and moxibustion group was slower than that in the control group after moxibustion intervention,P<0.05.There was no significant difference between the model group and the moxibustion group(P>0.05).After moxibustion intervention,compared with the control group,the weight gain slowly in the model group and the moxibustion group,P<0.05,and compared with the moxibustion group,the weight gained slowly in the model group,P<0.05.(3)The hind paw volume comparison:there was no siginicant difference between the 3 groups(P>0.05)before establishing the CIA model,and the baseline was comparable.After successful modeling,compared with the control group,the hind paw volume of the rats in the model group and the moxibustion group increased,P<0.05,and there was no difference between the model group and the moxibustion group(P>0.05).After moxibustion intervention,compared with the control group,the hind paw volume of the model group and the moxibustion group increased significantly(P<0.05),and compared with the model group,the hind paw volume decreased in the moxibustion group significantly(P<0.05).(4)Comparison of AI scores:in the model group and the moxibustion group,AI were more than 4 points.Compared with the model group,there was no significant difference in AI scores between the moxibustion group and the model group(P>0.05),and the baseline was comparable.After the intervention of the moxibustion,the AI scores of the moxibustion group was lower than in the model group significantly(P<0.05).(5)Inflammatory factors:the level of IL-1?:after the moxibustion intervention,conpared with the control group,the rat serum IL-1? increased significantly in the model group(P<0.05).Compared with the model group,the rat serum IL-1?decreased significantly(P<0.05).the level of TNF-?:after the moxibustion intervention,compared with the control group,the rat serum TNF-? increased in the model group significantly(P<0.05),and compared with the model group,the rat serum TNF-? decreased significantly in the moxibustion group(P<0.05).the level of PGE2:after the moxibustion intervention,compared with the control group,the rat serum PGE2 level in the model increased significantly(P<0.05),and compared with the model group,the rat serum PGE2 level in the moxibustion group decreased significantly(P<0.05).(6)The pathological tissue of the ankle joint of rats:the rats ankle joint tissue structure were clear,smooth surface,non synovial cell lesions,no inflammatory cell infiltration in the control group;the rats ankle joint tissue in the model group were synovial cell degeneration,synovial tissue fibrous tissue proliferation and inflammatory cell infiltration;the rats ankle joint tissue in the moxibustion group were only synovial thickening.(7)The serum metabonomics analysis:compared with the control group,the level of threonine,serine,glutamic acid,aspartic acid,leucine,isoleucine,taurine and phenylalanine increased significantly(P<0.05),and the level of citric acid decreased significantly(P<0.05).Compared with the model group,the level of serine,leucine,isoleuvine and taurine decrease in the moxibustion group,while the citric acid increased(P<0.05).The level of other metabolites decreased,but the difference has no statistically significant(P>0.05).(8)The articular synovium tissues metabonomics analysis:compared with the control group,the level of lysine,glycine,asparagine,aspartic acid,valine and phenylalanine in the model group increased significantly,the level of Petroselinic acid,cystathionine,cholesterol decreased significantly(P<0.05).Compared with the model group,the level of valine,glycine,asparagine,aspartic acid and phenylalanine decreased in the moxibustion decreased(P<0.05),and the level of cystathionine,cholesterol increased significantly(P<0.05).The level of other metabolites decreased,but the difference was no statistically significant(P>0.05).(9)16 metabolic pathways were involved in the screening of differential metabolites:Valine,leucine and isoleucine biosynthesis;D-Glutamine and D-glutamate metabolism;Glycine,serine and threonine metabolism;Phenylalanine,tyrosine and tryptophan biosynthesis;Alanine,aspartate and glutamate metabolism;Taurine and hypotaurine metabolism;Methane metabolism;Phenylalanine metabolism;Glyoxylate and dicarboxylate metabolism;Cysteine and methionine metabolism;Primary bile acid biosynthesis;Arginine and proline metabolism;Glutathione metabolism;Citrate cycle(TCA cycle);Steroid biosynthesis;Aminoacyl-tRNA biosynthesis.Conclusion:(1)In this study,moxibustion can improve CIA rats' body weight,hind paw volume and AI scores,and moxibustion can also improve the level of serum IL-1?,TNF-?,PGE2.The pathology of ankle joint in the moxibustion group only showed synovial thickening.Moxibustion in rats can improve joint function,reduce inflammation and had a therapeutic effect of regulating immune.(2)The metabolism of serum and articular synovium changed significantly after CIA modeling.The potential markers and their related metabolic pathways were screeded out which were associated with RA.(3)Moxibustion can restore the abnormal metabolites in the serum and articular synovium of CIA rats.The metabonomics can screen potential therapeutic targets of moxibustion. |
PDF Full Text Request