| ObjectiveRheumatoid arthritis(RA)is a serious systemic autoimmune disease that mainly affects peripheral joints,causing joint swelling and pain due to synovial hyperplasia and inflammatory cell infiltration,and eventually disability due to joint bone destruction.According to Chinese medicine,the kidney is the master of bone and marrow production,and kidney deficiency may be closely related to RA disease activity,disease progression,and prognosis.In patients with rheumatoid arthritis,the determination of disease activity has become an important component of RA management.The aim of the present study was to investigate the association between kidney deficiency and disease activity in RA patients,and to find useful biomarkers for the diagnosis of disease-syndrome in kidney deficiency type RA patients.MethodsSixty patients with RA were included and divided into kidney deficiency group and non kidney deficiency group according to the diagnostic criteria of TCM syndromes,and clinical data(gender,age,age of onset,disease duration,swollen joint count,tender joint count,erythrocyte sedimentation rate,C-reactive protein,platelet count,rheumatoid factor,anti-CCP,CDAI,SDAI,DAS28 score,HAQ score)were collected and compared between groups for Statistical analysis.Serum samples were selected from 13 RA patients and 8 healthy individuals from physical examination centers,and divided into RA kidney deficiency group,RA non kidney deficiency group and healthy control group.Protein quantification was performed by proteomics technique(BCA method)for each group of serum samples,followed by liquid chromatography-tandem mass spectrometry(LC-MS/MS)coupling technique for detection.The obtained mass spectrometry data were identified by searching in the human Fasta database using Maxquant software(version 1.6.17.0).The screened differential proteins were defined as differential proteins between groups when the fold of difference between groups was ≥1.5(up-regulation)or ≤0.7(down-regulation)and the test q-value was <0.05.Bioinformatics analysis uses data packages "ggplot2","Venn Diagram","Coplex Heatmap","RColor Brewer",and "corrplot" to analysis KEGG pathway,Gene ontology annotation,"pearson" cluster classification for the identified differential proteins." R language(version4.0.2)is used to program and draw gene expression heat maps and gene enrichment bubble maps.Results1.The study included kidney deficiency RA group(40 cases)and non kidney deficiency RA group(20 cases),with the same male to female ratio of 1:5.67 in both groups.The average age of onset in the kidney deficiency group was 44.66±11.01 years,with no statistically significant difference between the two groups(P>0.050).The duration of disease was longer in the kidney deficiency group,with a mean of 9.31 ± 7.26 years,and the difference between the two groups was statistically significant(P<0.050).2.The average inflammatory index ESR was 57.33±21.95 mm/h and the average CRP was 32.66±29.08 mg/L in the kidney deficiency group,which were elevated compared with the non kidney deficiency group,but the difference was not statistically significant when comparing the two groups at P>0.050.The average autoantibody RF titer in the kidney deficiency group was 383.03±449.48 IU/m L,which was significantly higher than that in the non kidney deficiency group(195.46±205.23 IU/m L),and the difference was statistically significant when comparing the two groups,P<0.050.The average anti-CCP in the kidney deficiency group was 132.69±77.34 U/m L,and the difference was not statistically significant when compared with the non kidney deficiency group.3.The clinical evaluation indexes and disease activity were higher in the renal deficiency group,and the differences between the two groups were compared in the swollen joint count(P=0.014),tender joint count(P=0.043),overall patient assessment(P=0.012),disease activity DAS28-CRP score(P=0.012),CDAI(P=0.010),SDAI(P=0.010),all of which were statistically significant,and DAS28-ESR score(P=0.007),HAQ(P=0.002)and overall physician evaluation(P=0.006)were all statistically significant(P<0.010).4.Protein quantification was performed on serum samples from 13 RA cases and 8healthy individuals,and a total of >1000 proteins were identified,and 241 proteins were significantly different compared to healthy controls.When analyzed in comparison with healthy controls,there were 96 differential proteins in the RA kidney deficiency group and50 were upregulated;111 differential proteins in the high disease activity group and 60 were upregulated;and only 76 differential proteins in the non kidney deficiency group.The gene IDs were used to indicate the screened differential proteins,and 49 of the 241 differential genes were rheumatoid arthritis target genes.The expression of SAA1,CRP,S100A9,SAA2,FGB,F9,MBL2,TNC,PRG4,and COMP were significantly up-regulated in the kidney deficiency group and high disease activity group,while the expression of TNXB,CAT,CA1,PRDX2,IGFBP5,GSN,ENPP2,and PON1 were significantly down-regulated,compared with the healthy control group and non kidney deficiency group.5.KEGG signaling pathway enrichment analysis showed that 241 differential genes were mainly enriched in complement and coagulation cascade reactions,extracellular matrix-receptor interactions,focal ligation,platelet activation,Staphylococcus aureus infection.6.GO annotation analysis of significantly up-regulated proteins in RA kidney deficiency group revealed that their related biological functions are mainly involved in biological processes such as platelet degranulation,acute phase response,complement activation,extracellular matrix organization,fibrinolysis,coagulation,protein hydrolysis,receptor-mediated endocytosis,innate immune response,platelet activation,etc.Molecular functions are mainly heparin binding,calcium binding,antioxidant activity,receptor binding and cell adhesion molecule binding.The down-regulated proteins are mainly involved in cell adhesion,triglyceride metabolic processes,and cholesterol metabolic processes.ConclusionKidney deficiency is closely related to RA disease activity,and patients with kidney deficiency present with more severe clinical signs and symptoms and disease effects.Kidney deficiency RA has more protein responses involved in signaling pathway regulation and biological processes,and may have more complex physiological-pathological immune response mechanisms and interactions.The up-regulated differential proteins SAA1,SAA2,S100A9,FGB,FGG,TNC,COMP,MBL2,PRG4 and the down-regulated differential proteins TNXB and CAT may be useful biomarkers for disease severity assessment and disease-syndrome diagnosis in kidney deficiency RA and may provide additional information about disease activity. |
PDF Full Text Request