Impact Of Inflammatory Cytokine IL-1? On The Expression Of Multiple Drug Resistance Protein P-gp In Peripheral Blood Lymphocytes Of Patients With Rheumatoid Arthritis

Background: Rheumatoid arthritis(RA)mainly affects the facet joints,and it has the characteristics of chronic destruction of symmetrical facet joints.It can lead to multiple joint structure and function impairment in the whole body at the late stage.So far,hormones,immunosuppressants and biologics are still the first choices for clinical treatment of RA.However,as the time of treatment is prolonged,many patients suffer from persistent symptoms or recurrent symptoms,which may be related to multidrug resistance(MDR).Among them,the family member P-glycoprotein(P-gp),which acts as an energy-dependent pump,can transport the drug out of effector cells.IL-1?,as a cytokine in the pathogenesis of RA,permeates the disease progression and aggravates the inflammatory response.Our previous studies have confirmed that in lymphocytes of RA patients,as the increase of concentration and the extension of time,the expression of P-gp changed accordingly under the influence of IL-6 and IL-17.Therefore,by observing the changes of p-gp on peripheral blood lymphocytes by the inflammatory cytokines IL-1?,we will explore P-gp-mediated multidrug resistance.Objective: To observe the effect of inflammatory cytokine IL-1? on the expression of multiple drug resistant(MDR)protein P-gp in peripheral blood lymphocytes of patients with Rheumatoid Arthritis(RA).Methods: Twenty newly diagnosed RA patients were enrolled,the peripheral blood of the above RA patients was collected,the serum was centrifuged,then we conducted research from cytokines,gene expression,and protein levels,the levels of IL-1? in group were quantified using Cytometric Bead Array(CBA).At the same time,we separated lymphocytes and established a system of training,then setted up blank control group and added different concentrations of IL-1? group,the mRNA levels of P-gp were detected by RT-PCR and P-gp function was detected by rhodamine 123 experiment(Rh123).Similarly,the mRNA levels and function of P-gp were measured again at different time.Results: 1.The levels of IL-1? in newly diagnosed RA patients were quantitatively analyzed by CBA:The levels of IL-1? in peripheral blood of different RA patients were compared with those of healthy individuals,the conclusion can be drawn that the levels of IL-1? in RA patients were higher than that in healthy people(P<0.1).There was no correlation between the levels of IL-1? and the expression of P-gp mRNA in RA patients(P>0.1).2.The mRNA levels of P-gp were detected by RT-PCR:Different levels of P-gp mRNA expression were observed under 0ng,5ng,10 ng of IL-1?(P<0.1).Further analysis was made on the difference of three concentrations at each time of 0h,24 h,48h,there were differences between different concentration treatments at the time level of 24 h and 48h(P<0.1).At both time levels above,5ng was higher than that in the 0ng group(P<0.1).The expression of P-gp mRNA was different with the prolongation of culture time by stimulation of IL-1?(P<0.1).At the same concentration level,it was increased at 24 h and 48h(P<0.1),and the most obvious increased in 48 h group(P<0.05).3.The P-gp function was detected by Rh123 experiment:Under the action of 0ng,5ng,10 ng of IL-1?,the Rh123 fluorescence intensity was not different(P>0.1).Under thestimulation of IL-1?,the Rh123 fluorescence intensity decreased significantly with the extension of cell culture time(P<0.1).At the same level of concentration,there was a decrease in the 48 h group at 5 ng(P<0.1)Conclusion: 1.The serum IL-1? in patients with newly diagnosed RA was higher than that in normal people.There was no significant correlation between the expression of IL-1? and P-gp mRNA.2.Under the influence of specific concentration and time of IL-1?,the expression of P-gp mRNA in peripheral blood lymphocytes of newly diagnosed RA patients was significantly increased and there was a certain degree of concentration and time correlation,which provided a basis for exploring multidrug resistance induced by P-gp.3.Under the influence of specific concentration and time of IL-1?,the function of P-gp in peripheral blood lymphocytes of newly diagnosed RA patients was significantly enhanced.At the same time,the relationship between the two showed a certain degree of concentration and time correlation.Similarly,the theoretical basis for exploring multidrug resistance under the action of P-gp was made.