MiR146a In The Pathogenesis Of Rheumatoid Arthritis And Its Correlation With Traditional Chinese Medical Constitutions

BackgroundRheumatoid arthritis(RA)is characterized by synovitis and bone destruction and can also affect multiple organs.The pathogenesis of RA is to be studied;Multiple studies suggest that miR-146a may participate the pathological process of RA,but the related mechanism needs to be further studied;Genetic polymorphism is an important internal factor of medical constitution,and the study of the association between genetic polymorphism and medical constitution is helpful for the judgment of disease-specific susceptibility;Genetic polymorphism of miR-146a sequence rs2910164 in RA patients and its correlation with TCM medical constitution are unclear.Objective1.Reveal the expression of miR-146a in RA patients,its correlation with the disease activity and its regulation on pro-inflammatory cytokine expression to help clarify of the pathogenesis of RA.2.Study the polymorphism of the rs2910164 locus of miR-146a sequence in RA patients,and the relationship with the constitution of Chinese medical constitution,to enrich the connotation of the theory of Chinese medicine constitution.MethodsAccording to the inclusion exclusion criteria,patients with RA and healthy controls were included.The clinical data of patients were collected.Real-time PCR was used to detect the expression of miR-146a in PBMCs and urine sediment.Correlation between the expression of miR-146a and the clinical characteristics of RA were analyzed.Transfection of miR-146a mimic were conducted to determine the effect of miR-146a on the expression of pro-inflammatory factors in PBMCs from RA patients.HRM allele typing kit was used to detect the polymorphism at the rs2910164 locus of miR-146a sequence.According to the standard of TCM Constitution Classification and Judgment,classification of TCM constitution of RA patients were determined;Differences of Chinese medical constitution of RA patients in different genotypes at rs2910164 were analyzed.Statistical analysis using SPSS 20.0 software,P<0.05 for the difference was statistically significant.Results1.The expression of miR-146a in PBMCs was significantly higher in RA patients than in healthy controls(P<0.05).The expression levels of miR-146a was higher in active RA patients than in stable RA patients(P<0.05).The expression levels of miR-146a was higher in RA patients with extraarticular manifestations than in RA patients without extraarticular manifestations(P<0.05).The expression of miR-1 46a in PBMCs was positively correlated with disease activity score in 28 joints(DAS28),ESR and CRP,but not correlated with disease duration,RF and anti-CCP antibody(P<0.05).The expression of miR-146a in urinary was significantly higher in RA patients than in healthy controls(P<0.05).The expression levels of miR-146a was higher in active RA patients than in stable RA patients(P<0.05).The expression levels of miR-146a was higher in RA patients with extraarticular manifestations than in RA patients without extraarticular manifestations(P<0.05).The expression of miR-146a in urinary was positively correlated with disease activity score in 28 joints(DAS28),ESR and CRP,but not correlated with disease duration,RF and anti-CCP antibodies(P<0.05).2.Transfection of miR-146a mimic significantly reduced the mRNA expression levels of TNF-α,IL-1β and IL-6 in PBMCs of RA patients(P<0.05).3.The proportion of patients with GC genotype at the rs2910164 locus of miR-146a has significantly increased extra-articular manifestations(P<0.05);The expression level of miR-146a in urine sediment of RA patients with G/G genotype is significantly lower than that with G/C or C/C genotype(P<0.05);Serum levels of TNFa in patients with RA with G/C genotype were significantly higher than those in patients with RA with G/G and C/C genotype(P<0.05).4.The proportion of blood stasis in women with RA is significantly higher than that in men(P<0.05);The proportion of yang deficiency in active RA is significantly higher than that in inactive RA,while the proportion of RA patients in active stage is generally lower than that in inactive RA(P<0.05);The proportion of RA with extra-articular clinical manifestations of yang deficiency is significantly higher than that without extra-articular clinical manifestations,while the proportion of patients with extra-articular clinical manifestations of RA and mild RA is significantly lower than that without extra-articular clinical manifestations performance RA(P<0.05);The most common TCM constitution type in RA patients with G/G genotype is blood stasis,while the most common TCM constitution in RA with G/C genotype and C/C genotype type is Yang deficiency quality.Conclusion1.MiR-146a has abnormal expression in PBMCs and urine sediment of RA patients,and may participate in the pathological mechanism of RA by affecting the expression of proinflammatory cytokines in PBMCs.2.The polymorphism at the rs2910164 locus of miR-146a can affect the expression of miR-146a and the level of TNFa in RA patients,and is correlated with the extra-articular clinical manifestations of RA.3.The type of Chinese medical constitution of RA patients is related to specific clinical manifestations,and may be related to the rs2910164 polymorphism of miR-146a.