Regulation Of TRAIL On P-gp Expression In Peripheral Blood Lymphocytes Of Children With Steroid Resistant Asthma

ObjectiveTo explore the mechanism of Tumor necrosis factor-related apoptosis-inducing ligand reversal steroid resistance mediated by P-glycoprotein(P-gp),To explore new therapeutic targets for steroid resistance in children with asthma.MethodsA total of 46 children with Uncontrolled asthma(UA)and children with Controlled asthma(CA)were selected from our department of pediatrics from September 2020 to December 2022.Among them,36 patients was uncontrolled asthma and 10 patients was controlled asthma.13 healthy subjects were selected as control group.On the first day after admission,3 m L of fasting venous blood was collected from all the children,and peripheral blood lymphocytes were extracted after anticoagu Lant centrifugation.1.P-gp expression levels in peripheral blood lymphocytes of CA,UA and healthy controls were detected by enzyme-linked immunosorbent assay(ELISA)and flow cytometry.2.The imported lung function instrument was used to measure the lung function parameters of the tested children according to international standards:Forced expiratory volume in the first second(FEV1),Pearson test was used to analyze the correlation between P-gp expression level and FEV1 index in children with asthma.3.TRAIL expression levels in CA and UA plasma were detected by enzyme-linked immunosorbent assay(ELISA).4.The lymphocyte of UA children group was treated with different concentrations of TRAIL(0 ug/L,1 ug/L,10 ug/L,100 ug/L)for 48 h.P-gp m RNA expression level was detected by polymerase chain reaction(PCR).5.Western Blot(WB)was used to detect the expression level of P-gp protein in peripheral lymphocytes of children with UA group before and after 10ug/L TRAIL intervention.ResuLts1.According to ELISA and flow flow resu Lts,the expression level of P-gp in lymphocytes in UA group was higher than that in CA group and healthy control group,and the difference was statistically significant(P<0.05).2.According to the flow flow results,the percentage of lymphocyte P-gp positive in UA group was higher than that in CA group and healthy control group,and the difference was statistically significant(P < 0.05).3.There was a negative correlation between the expression of P-gp in lymphocyte and lung function(FEV1/FVC)in UA group.(r=-0.4637,P < 0.05).4.The plasma TRAIL expression in UA group was lower than that in CA group,and the difference was statistically significant(P<0.05).5.After intervention with exogenous TRAIL concentrations of 1 ug/L,10ug/L and 100 ug/L,the expression level of P-gp m RNA in lymphocytes in UA group was significantly decreased,with statistical significance(P<0.05).When10 ug/L TRAIL was used,the down-regulation effect on P-gp was the most obvious.6.After 10 ug/L exogenous TRAIL intervention,P-gp protein expression in lymphocytes of children in UA group was decreased,with statistical significance(P<0.05).Conclusions1.P-gp was highly expressed in lymphocytes of children with uncontrolled asthma,and was negatively correlated with FEV1 indexe of pulmonary function in children with asthma,suggesting that P-gp was involved in the formation of steroid resistance mechanism in children with asthma,which could be used as a reference for evaluating the therapeutic effect of asthma2.The TRAIL showed low expression in plasma of children in UA group,TRAIL can reverse steroid resistance and restore drug sensitivity in asthmatic children by down-regulating P-gp expression.