The Correlation Of Expression Of Differential Drug-resistant Proteins And Inflammatory Cytokines In Collagen-induced Arthritis Model

Objective:By comparing the differential expression of ATP-binding cassette?ABC?transporter family resistance-related proteins P-gp,BCRP,MRP1 and inflammatory cytokines IL-1?,IL-2,IL-6,IL-10,TNF-?and IL-17 in CIA model mice,to investigate the correlation between Inflammatory cytokines and drug-resistant proteins.Methods:Fourteen DBA1 mice were successfully induced by collagen and Freund's Adjuvant.According to the scores of synovial pathology,the CIA group was divided into mild,moderate-to-severe groups,another four mice were selected as control.The mRNA expressions of P-gp,BCRP,MRP1 in splenic lymphocyte cells were measured by reverse transcription-polymerase chain rection?RT-PCR?.The concentrations of IL-1?,IL-2,IL-6,IL-10,TNF-?,IL-17 in serum were detected by Cytometric Bead Array?CBA?.Further analyzed the correlation between different inflammatory cytokines and these proteins,then studiedoneof the proteinswhichwasmost relatedwithcytokinesby immunohistochemical?IHC?in synovium.Results:1.Among the three groups,compared with the normal controls[7.75?5.14,9.17?],the level of IL-6 in the serum of mild CIA group[25.31?15.15,29.27?],moderate-severe CIA group[45.03?38.87,64.02?]were significantly increased?Z=14.383,P₁<0.05?,compared with the mild CIA group,the level of IL-6 in serum were significantly increased in the moderate-to-severe CIA group?Z=-7,P<0.05?.Among the three groups,compared with the normal controls[22.81?20.84,28.17?],the level of TNF-?in the serum of mild CIA group[45.00?32.76,58.51?],moderate-severe CIA group[45.00?39.78,58.95?]were significantly increased?Z=8.375,P<0.05?,there was no distinct difference between the mild CIA group and the moderate-to-severe CIA group?Z=-0.381,P>0.05?.There was no significantly difference in the IL-1?,IL-2,IL-10,IL-17 levels.2.In the spleen lymphocytes,there was no significant difference in the the mRNA expression level of P-gp and BCRP among the groups,but the mRNA expression level of MRP1 were significantly increased?Z=12.634,P<0.05?.Compared with the mild CIA group,the MRP1 mRNA in the moderate-to-severe CIA group was higher,there was significant difference?Z=-2.582,P<0.05?.There was a correlation between mRNA expression of MRP1 and P-gp?r=0.635,P=0.015?.3.The mRNA expression of MRP1 was positively correlated with IL-6 level?r=0.711,P=0.004?.4.The expression of MRP1 in normal group,low-level IL-6 group and high-level IL-6group were respectively as follows::1.08?0.65,1.30?,1.40?1.13,2.07?,1.90?1.68,2.17?.5.Compared with the controls,the cytoplasm/membrane of the ankle joint synovial tissue in the mild CIA group was yellow,which proved that MRP1 expression was weakly positive.It is yellowish-brown in the moderate-to-severe CIA group,which proved that MRP1 expression was strongly positive.Conclusion:In the CIA arthritis model,synovial tissue lesion is not only related to inflammatory cytokines,but also related to MRP1 expression in the ABC transport resistance protein family,and it is proved that IL-6 is highly correlated to MRP1.