Impact Of Inflammatory Cytokine IL-1? On The Expression Of Multiple Drug Resistance Protein BCRP/ABCG2 In Synoviocytes Of Patients With Rheumatoid Arthritis

Background:Rheumatoid arthritis(RA)is an autoimmune disease caused by inflammation of the synovium and characterized by chronic polyarthritis that destroys bone and cartilage,which has a high disability rate.So far,commonly used drugs to treat RA were clinically,glucocorticoids,modified anti-rheumatic drugs(traditional DMARDs,mostly immunosuppressants),and biological agents DMARDs.However,many patients continue to have no effective effect or recurrent symptoms after long-term use,which is closely related to multidrug resistance(MDR).The study found that the most critical mechanism for the production of MDR is the ABC transporter family-mediated increase in target cell drug excretion,and inflammatory cytokines play a role in this process.Our previous research results have confirmed that interleukin(IL)-1?,IL-6 and IL-17 are involved in the regulation of P-glycoprotein(P-gp)expression in lymphocytes of RA patients,which has certain Time and concentration dependence.The breast cancer resistance protein(BCRP/ABCG2),the first ABC semi-transporter found in the cell membrane,is also a highly efficient efflux pump,which absorbs,distributes,and excretes drugs or exogenous substances in the body.IL-1? is a cytokine involved in the whole process of RA,which can aggravate the inflammatory response.Fibroblast-like synoviocytes(FLS)and various inflammatory cells or cytokines interact to participate in the occurrence and development of RA.Therefore,we observed the regulation of IL-1? on the multiple drug resistance protein BCRP/ABCG2 in RA-FLS,and explored the mechanism by which BCRP induces the generation of MDR.Objective:To explore the effects of different concentrations or time points of IL-1? on the expression of BCRP mRNA in FLS of RA patients.Methods:Synovial tissues of RA and Osteoarthritis(OA)patients(n=3)were extracted by means of Joint surgery,FLS culture system in vitro was established.After FLS passaged to the fifth generation,we did the next experiments.1)Compared expression levels of BCRP mRNA in RA-FLS and OA-FLS by PCR.2)We collect clinical and laboratory data of these patients and calculate the disease activity score(DAS28).3)FLS cells were divided into A,B,C,D four groups randomly with different treatments: Group A was the control group.Group B,C,D were respectively co-cultured with low(20ng/ml),middle(50ng/ml),high(100ng/ml)concentrations of IL-1? for 24 hours,48 hours,and 72 hours.The effect of IL-1? on the expression level of BCRP mRNA was observed at different concentrations and time points.Results:1.The expression of BCRP mRNA in FLS of RA patients was significantly higher than that in OA group(P<0.05).2.The expression of BCRP mRNA in FLS of RA patients was positively correlated with disease course,erythrocyte sedimentation rate,C-reactive protein,and DAS28,but negatively correlated with age of onset,and had no correlation with gender.3.The expression of BCRP mRNA in RA-FLS stimulated by the same concentration of IL-1? at different time conditions: there were differences in the concentration of IL-1? at 0ng/ml,20ng/ml and 100ng/ml(P<0.05).While the concentration of IL-1? was 0 ng/ml,the expression of BCRP mRNA in the 48 hours and 72 hours groups was higher than that in the 24 hours group(P<0.05);Under the condition,which the concentration of IL-1? was 20ng/ml or 100ng/ml,the expression of BCRP mRNA in the 72 hours group was significantly higher than that in the 24 hours group(P<0.05).4.The expression of BCRP mRNA in RA-FLS stimulated by different concentrations of IL-1? at the same time: when the stimulation time was 72 hours,the expression of BCRP mRNA in the 100ng/ml group was higher than that in the 0ng/ml group(P<0.05).Conclusion:1.Compared with patients with OA,the expression of BCRP mRNA in FLS of RA patients was significantly increased,which suggests the presence of BCRP-mediated MDR in RA.2.The expression of BCRP mRNA in FLS of RA patients may be correlated with disease activity.3.The stimulation time and concentration of IL-1?were correlated with the expression level of BCRP mRNA in FLS of RA patients,and the time dependence was more significant than the concentration-dependent.This suggests that IL-1? is involved in BCRP-mediated multidrug resistance and lays the foundation for exploring its specific mechanism.