Correlation Between NF-?B Signaling Pathway And Resistance To Methotrexate In Patients With Rheumatoid Arthritis And TCM Syndromes

Objective This study was detected the expression or function of nuclear factor-kappa B(NF-?B)signal pathway's related protein?gene and downstream inflammatory factors,such as NF-?B?inhibitor of nuclear factor kappa-B(I?B)?multidrug resistance 1(mdrl)?P-glycoprotein(P-gp)?Tumor Necrosis Factor-?(TNF-?)?Interleuikn-1?(IL-1?)in the normal and rheumatoid arthritis(RA)patients(sensitive to methotrexate treatment and resistance to methotrexate treatment).Our purpuse was to explore the possible mechanism of NF-?B signal pathway in MTX resistance and the relevance to traditional Chinese medicine Syndrome(TCM Syndrome),providing new ideas and targets to improve RA treatment,overcoming MTX resistance.Method Following the inclusion criteria,we gathered RA patients and healthy controls,and then according DAS28 score based on MTX effects dividied RA patients into MTX sensitive group and resistant group.Using ELISA to detect the concentration of NF-?B?I?Ba?TNF-??IL-1?,adopting real-Time PCR to detect the expression of mdr1 mRNA?TNF-amRNA?IL-1?mRNA,Flow Cytometry(FCM)was used to detect the expression and function of P-gp.Making a comparison in the expression or function of NF-?B pathway related protein gene and downstream inflammatory factors and the TCM syndromes correlation in each group.Results1.The concentration of NF-?B?I?B? in serum:Compared with normal control group,the concentration of NF-?B was higher in MTX sensitive group and MTX resistance group,the difference was statistically significant(P<0.05);there was also a increased expression of NF-?B in MTX resistance group compared with MTX sensitive group(P<0.05).Compared with normal control group,the concentration of I?B? was lower than MTX sensitive group and MTX resistance group,the difference was statistically significant(P<0.05);there was also a reduced expression of I?B? in MTX resistance group compared with MTX sensitive group(P<0.05).2.The expression and function of P-gp in peripheral blood monouclear cells:The expression and function of P-gp in MTX sensitive group and MTX resistance group were higer than normal control group,the difference was statistically significant(P<0.05);Compared with MTX sensitive group,the expression and function of P-gp in MTX resistance group increased with significant difference(P<0.05).3.The level of mdrl mRNA?TNFa mRNA?IL-1 ? mRNA in peripheral blood monouclear cells:Compared with MTX sensitive group,the expression of mdrl mRNA?TNF? mRNA?IL-1 ? mRNA were higher in MTX resistance group,the difference was statistically significant(P<0.05).4.The concentration of TNF-??IL-1? in serum:TNF-??IL-1? expression in MTX sensitive group and MTX resistance group were higher than normal control group,which possess significant difference(P<0.05).In contrast with MTX sensitive group,the concentration of TNF-??IL-1? were increased in MTX resistance group,the difference was statistically significant(P<0.05).5.The relationship between TCM syndrome types and MTX drug resistance:The distribution of TCM syndrome types(the syndrome of dampness-heat blocking collaterals group?the syndrome of cold-dampness blocking collaterals group?the syndrome of intermingled phlegm and bloods?liver and blocking deficiency syndrome)in MTX sensitive group and MTX resistance group were not statistically significant(P>0.05).There was also no statistically significant difference of the expression levels of NF-?B?I?B??TNF-??IL-1??mdrl mRNA?TNF-? mRNA?IL-1 ? mRNA among all the syndromes(P>0.05).Conclusion1.The high expression of NF-?B signal pathway and downstream inflammation factor can activate the mdrl,raised P-gp expression and fuction,and is associated with MTX drug resistance.NF-?B signaling pathway activation may be one of the forming mechanism of RA patients with MTX drug resistance.2.MTX drug resistance and TCM syndrome types have no obvious correlation.