Study On Rheumatoid Arthritis Pharmacodynamics And Metabolomics Based On Atractylodes Macrocephala

1PURPOSERheumatoid arthritis(RA),which is clinically called "bi" in traditional Chinese medicine,with a high incidence and a low cure rate.There are a large of Chinese-western medicine preparations,and side effects was extremely,and the effect of therapeutic needs to be improved.As technology advances,scholars have paid more attention to finding medicines or prescriptions for treating “bi” from classic Chinese medicine books.Atraclylodes lancea is an aromatic wet medicine,which has the effects of dampness and spleen,driving wind and scattering cold,and can be used for abdominal fullness,rheumatism and pain.At present,the research on Atraclylodes lancea mainly focuses on the effects of "dryness and dampness and strengthening the spleen",and there are few reports on the use of "dryness" to treat "bi".In this study,we used Atraclylodes lancea as the subject,and screened the main pharmacodynamic sites for RA through in vivo pharmacodynamic studies,and verified their effects using cell models;further applied GC-MS(gas chromatography-mass spectrometry)technology and related analytical methods The metabolic pathways of Atraclylodes lancea for the treatment of rheumatoid arthritis were preliminarily predicted,which provided a reference basis for the effectiveness of Atraclylodes lancea for the treatment of rheumatoid arthritis,and laid the foundation for the further development of new anti-rheumatoid arthritis drugs with strong efficacy and small side effects.2METHOD2.1Screening of effective parts of Atraclylodes lancea for the treatment of RAWistar rats were grouped into 7 groups: normal group,model group,Atractylodes volatile oil high and low dose group,Atractylodes alcohol extract high and low dose group,positive drug group,AIA(adjuvant arthritis)model was created in addition to the normal group,behavioral changes,the foot pad thickness and joint scores changes,and serum IL-6 and IL-17 content as evaluation indicators,screen the main medicinal parts of Atraclylodes lancea for RA.2.2 Pharmacodynamics of Atraclylodes oil in vivo and in vitro for RAWistar rats were grouped into 5 groups: normal group,model group,Atractylodes oil high and low dose group,positive drug group,AIA(adjuvant arthritis)model was created in addition to the normal group,the scores of joint changes,serum IL-6,IL-17,TNF-?,and IL-1? content,ankle joint HE sections,and immunohistochemical MMP2 and MMP9 contents were used as evaluation indicators to investigate the anti-RA efficacy of Atractylodes oil.Choose the rheumatoid arthritis synovial fibroblasts(MH7A)as research objects for in vitro experiments,TNF-? induced MH7 A model was administered with different concentrations of Atractylodes oil,Measurement of cytotoxicity and cell proliferation experiments.Investigate the in vitro pharmacodynamic effect of Atractylodes oil in treating RA.2.3Analysis of Metabolomics of Atractylodes Oil for RA by Metabolomics based on GC-MSUse the GC-MS technology to analyze the serum components of normal,model and high-dose Atraclylodes oil group in " Pharmacodynamics Study of Atractylodes Oil for RA".The serum metabolite map was established and analyzed by PCA,PLS,OPLS,find the difference metabolites between the groups,use Metabo Analyst analysis method to analyze the metabolic pathways of the different components,and predict the possible metabolic pathways of Atractylodes oil to play a role in treating RA.3RESULT3.1 Screening of effective parts of Atraclylodes lancea for the treatment of RACompared with the normal group,rats in the right hind paw showed significant redness and swelling,and the serum IL-6 and IL-17 content significantly increased(P<0.05).After treatment,the redness and swelling of the paw in the positive drug group improved,and the serum IL-6 and IL-17 content decreased significantly(P<0.05);The degree of redness and swelling of the feet of rats in the high and low dose groups of Atraclylodes volatile oil significantly improved,and the serum IL-6 and IL-17 content significantly decreased(P<0.05).However,the symptoms of foot and paw of rats in high and low dose Atraclylodes alcohol extract group were not significantly improved,and there was no significant change in serum IL-6 and IL-17 content(P> 0.05).3.2 Pharmacodynamics of Atraclylodes oil in vivo and in vitro for RALiken to the normal group,the right hind paw of rats showed significant redness and swelling,and the serum IL-6,IL-17,TNF-?,and IL-1? content were significantly increased(P<0.05).After treatment,the right hind paw of the positive drug group rats improved,and the serum IL-6,IL-17,TNF-?,and IL-1? content decreased significantly(P<0.05).In the high-dose group of Atraclylodes oil,the paw of rats significantly improved,and the serum IL-6,IL-17,TNF-?,and IL-1? content significantly decreased(P<0.05).The redness and swelling of the paw in the low-dose group of Atraclylodes oil improved,and the serum TNF-?,IL-1?,and TNF-?content decreased significantly(P<0.05);Ankle HE section results illustrated that the model group had obvious inflammatory cell infiltration,narrowed joint space and neoplasia,and the Atraclylodes oil high dose group significantly improved after administration;The joint immunohistochemistry results illustrated that the high dose of Atraclylodes oil can suppression the increase of MMP2 and MMP9 expression content in cartilage synovium of AIA rats.Cell experiments showed that the cytotoxicity of Atractylodes oil is low,and the toxicity is less than 80?g · m L-1,and it significantly inhibits the abnormal proliferation of MH7 A cells induced by TNF-?.3.3 Analysis of Metabolomics of Atractylodes Oil for RA by Metabolomics based on GC-MSPCA,PLS and OPLS analysis results illustrate that the normal group and the model group can be well separated,indicating that there are obviously differences in serum metabolites between the normal and the model group;and also in the normal,model,and Atraclylodes oil group rats,28 common endogenous metabolites were identified in the serum,12 of them were obviously different,Introduce 12 differential metabolites into Metabo Analyst3.0 analysis platform,and the analysis revealed 6 related to the treatment of RA by Atraclylodes oil.Metabolic pathways are: arginine and proline metabolism;alanine,aspartic acid and glutamic acid metabolism,D-glutamine and D-glutamic acid metabolism,aminoacyl-t RNA biosynthesis,Galactose metabolism,taurine and hypotaurine metabolism.4CONCLUSIONThis study determined in vivo pharmacodynamic experiments that the main medicinal effect of Atraclylodes for RA is volatile oil;and integrated in vitro cell experiments,the good therapeutic effect of Atractylodes oil on RA was further evaluated;GC-MS and related analysis were used to preliminarily judge the 6 amino acids and metabolic pathways related to the treatment of RA by Atractylodes oil.It supplies a reference to the clinical application of Atractylodes fortunei and supplies potential Chinese medicine to the development of rheumatoid arthritis medicines.