| Background:Rheumatoid arthritis(RA)is a heterogeneous,systemic,autoimmune disease characterized by symmetrical polyarthritis.Heterogeneity means that the patient's genetic background is different,and the cause may not be single,so the pathogenesis is not the same.RA Chinese medicine belongs to the category of sickness",and the"Clinical Guide to Medical Treatment"points out:"The singer,the closed but not the same,the righteousness is evil,the visceral meridians can not be smooth,all suffer from loss of blood and blood,and carelessness,The wind,cold and dampness can be taken up by the virtual,and the stagnation is caused by the stagnation of the sputum.The current pathogenesis of RA is not very clear.Treatment,and toxic side effects,and moxibustion as a kind of external treatment of traditional Chinese medicine,with a holistic,simple and fast,small side effects and other advantages.Moxibustion can warm the collaterals,promote blood circulation and remove phlegm to treat snoring.Although moxibustion is a commonly used traditional Chinese medicine treatment for RA,its mechanism of action is not clear.In the early stage,our research group conducted research on moxibustion treatment of RA rats based on proteomics and serum metabolomics.The results showed that there were changes in protein and serum metabolites.The results of the study laid the foundation for further exploration of the mechanism of action of moxibustion in the treatment of RA.Compared to proteomics,metabolomics has the unique advantage of being able to perform high-throughput analysis of metabolites and to amplify small changes in protein levels on metabolites,making them easier to detect.The most commonly used samples for metabolomics are blood and urine.Compared with blood samples,urine samples have the advantages of more stable biological composition and relatively simple extraction steps.At the same time,due to the high throughput and large-scale nature of metabolomics,As a result,single serum samples can not meet the research needs,and blood metabolomics data and urine metabolomics data can complement each other and verify.Therefore,this study used urine metabolomics technology to analyze the mechanism of moxibustion treatment of collagen-induced arthritis ratsObjective:To explore the changes of urine metabolic profile of rats with collagen-induced arthritis(CIA)by using metabolomics(gas chromatography-mass spectrometry)to explore the mechanism of actionMethods:Twenty-four SPF male Sprague-Dawley rats were randomly divided into control group,model group and moxibustion group,with 8 rats in each group.The model group and the moxibustion group all underwent CIA rat model.After successful modeling,the moxibustion group performed moxibustion on both sides of"Jinshu"and"Zusanli"points,once a day,each acupuncture point was 10 min,for 6 consecutive days as a course of treatment,a total of 3 courses of moxibustion,treatment room One day rest;the control group and the model group were not treated with moxibustion,but the rats were fixed at the same time by the same method.In order to evaluate the incidence of RA rats and the efficacy of moxibustion treatment of RA,the body weight,left and right toe volume of rats before and after successful modeling(ie before moxibustion),moxibustion for 1 week,2 weeks,3 weeks.The mean value was measured;the arthritis index(AI)was measured before moxibustion,moxibustion for 1 week,2 weeks,and 3 weeks;histopathological results were analyzed after moxibustion.The urine samples of rats in each group were collected for 1 week,2 weeks and 3 weeks.The GC-MS analysis technique was used to explore the changes of urine metabolism spectrum in different treatment courses of RA and the three courses of moxibustion for urine metabolites.VarietyResult:(1)General situation:Before the model was established,the rats in the three groups were generally in good condition;before the moxibustion,the rats in the control group were generally in good condition,and the rats in the model group and the moxibustion group showed a decrease in the color of the fur,and the joints were red and swollen;After 3 weeks,there was no abnormality in the control group.There was no change in diet,activity and fur color in the model group.The joint redness and swelling were obvious.The joint redness in the moxibustion group was significantly improved,which was more active than the model group(2)Body weight:Before modeling,there was no significant difference in body weight between the three groups(P>0.05),and the baseline was comparable.Before moxibustion,compared with the control group,the body weight of the model group and the moxibustion group were statistically significant.Learning differences(P<0.05);moxibustion intervention for 1 week,2 weeks,compared with the control group,the weight of the model group and moxibustion group still had statistical difference(P<0.05);moxibustion intervention for 3 weeks,and compared with the control group,the body weight of the model group was statistically different(P<0.05).There was no significant difference in the body weight of the moxibustion group(P>0.05).(3)Toe volume:Before the model was established,there was no significant difference in the mean volume of the left and right toes in the three groups(P>0.05),and the baseline was comparable;before moxibustion and moxibustion for 1 week,compared with the control group.The volume of the toes in the model group and the moxibustion group increased significantly,and there was a statistical difference(P<0.05).There was no significant difference in the toe volume between the model group and the moxibustion group(P>0.05).Moxibustion intervention for 2 weeks and 3 weeks,compared with the control group,the toe volume of the model group and the moxibustion group was statistically different,and the toe volume between the model group and the moxibustion group was also statistically significant difference(P<0.05).(4)Al score:Before moxibustion,the Al scores of the model group and the moxibustion group were higher than 4 points.There was no significant difference between the model group and the moxibustion group(P>0.05),suggesting that the baseline was comparable;there was no significant difference in AI scores between the model group and the moxibustion group(P>0.05),but the AI score of the moxibustion group was significantly smaller than that of the model group;moxibustion 2 weeks and 3 weeks,the moxibustion group and the model group were large.The rats showed a significant decrease in AI scores(P<0.01).(5)Histopathology:HE staining of the ankle joints of the three groups showed no abnormalities in the ankle joint of the control group;the model group showed joint stenosis,severe congestion and edema,and a large number of inflammatory cell infiltration and tissue necrosis.In the moxibustion group,the joint synovial inflammation was alleviated,the joint capsule cavity was smooth,the structure level was clear,and there was mild congestion and edema.(6)Urine metabolomics analysis:GC-MS-based urine metabolomics describes the regulation of urinary metabolites during different intervention periods of rheumatoid arthritis.After 1 week of moxibustion intervention,the model group compared with the control group,Nicotinic acid,Pyruvate,Hippuric acid,Xanthine,Palmic acid,Stearic acid,Oleic acid,Glycolic acid,2-methylhippuric acid,L-Valine,urinary adenine,5 The concentration of-hydrosy-6-methoxyindole glucuronide and Spermidine increased,and the difference was statistically significant(P<0.05).The concentrations of L-Phenylalanine and Xanthurenic acid decreased,and the difference was statistically significant(P<0.05).After moxibustion treatment,compared with the model group,the 15 metabolite levels in the moxibustion group were all adjusted,but there was no statistical differenceAfter 2 weeks of moxibustion intervention,the model group was compared with the control group,the concentration of Malonic acid,Succinate,Indole sulfate,L-Glutamine,p-cresol increased,5-Hydroxyindoleacetic acid,4-Hydroxybenzaldehyde,Vanillylmandelic acid,Kynurenic acid,Phenylalanine,Uric The concentration of acid decreased(P<0.05).The concentration of Kynurenic acid,L-Phenylalanine,Uric acid,Malonic acid,Succinate and L-Glutamine in the moxibustion group was statistically significant(P<0.05).The concentration was significantly adjusted.The rest of the metabolite levels changed,but there was no statistical differenceAfter 3 weeks of moxibustion intervention,the concentration of Malonic acid,p-cresol sulfate,L-Alanine,3-methyldioxyindole,P-cresol glucuronide,Oxoglutaric acid was increased in the model group compared with the control group,Citric acid,dihydroxyquinoline,3-hydroxyanthranilic acid,The concentration of L-Phenylalanine decreased(P<0.05).Compared with the model group,the concentration of Citric acid,L-Phenylalanine,p-cresol sulfate,3-methyldioxyindole and Oxoglutaric acid in the moxibustion group was statistically significant(P<0.05),and the concentration was significantly adjusted.The other metabolite levels were changed.But there is no statistical difference(7)Metabolic pathway:Thr-ough analysis of diff'erential metabolites,moxibustion treatment of RA rats mainly involves metabolic pathways such as glycolysis/gluconeogenesis and TCA cycle,phenylalanine metabolism,tryptophan metabolism,Butyric acid metabolism,and purine metabolism.Conclusion:(1)Moxibustion intervention for 3 weeks can significantly improve the general condition,body weight,toe swelling and AI score of CIA rats,and can also reduce joint synovial inflammation and clear structure,indicating that moxibustion has the effect of improving arthritis(2)The mechanism of action of moxibustion to improve arthritis is related to the regulation of glycolysis/gluconeogenesis and TCA cycle,phenylalanine metabolism,tryptophan metabolism,purine metabolism,butyric acid metabolism. |
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