Flow Cytometric Evaluation Of Glucocorticoid Receptor In Steroid Response And Ginsenosides Efficacy In Clinical

Objective:To evaluate Glucocorticoid receptor in steroid-resistant and -sensitive patients with systemic lupus erythematosus(SLE) and nephrotic syndrome(NS) and ginsenoside(GS) efficiency on GR in SLE by FCM using a monoclonal antibody(mAb) and FITC-Dex probes.Method:To compare FCM analysis of GR with Mo-Ab and FITC-Dex with Radioligand Binding Assay(RLBA) and Western-blot and observe the allocation of two probes in U937 and K562 cells with or without Dex treatment.In order to assess the efficacy of this method in predicting steroid response,GRs ofT lymphocytes(CD3~+) and mononuclear cells(CD14~+) in 35 SLE patients and 23 NS patients were studied before treatment. Disease activity scores were determined before and after treatment and used to divide patients into 2 group:steroid-resistant and steroid-sensitive.Then GR's characters in two group were observed.And to evaluat the effect on GR and efficacy in clinical of GS,32 SLE patients were studied.Results:Positive correlation between FCM-FITC-Dex assay and radiometrie GR or FCM-mAb-GR and Western-blot analysis in U937 cells and K562 cells and their Dex treatment groups respectively.GR staining with anti-GR mAb and FITC-Dex were localized not only in cell nucleus but also cell cytoplasm.Determined by FCM,the expression and the binding of CD3/GR and CD14/GR in steroid-resistant patients with SLE were all lower than those in steroid-sensitive group,which did not differ from the results of normal controls. Similar results were found in NS patients.The binding of CD3/GR and CD14/GR in steroid-resistant NS were both lower than those in steroid-sensitive group,which did not differ from those of normal group.However,there were no significant differences in GR expression between steroid-resistant patients and steroid-sensitive patients with NS.Compared with placebo,GS couled upregulate GR binding by FCM detectation.Conclusion:Monitoring the GR with a GR-mAb and FITC-Dex probes by FCM would be useful and convenient in determination of GR in clinical and experiment.GR in steroid-resistant patients were lower than that in steroid-sensitive ones which may be one of indexes in steroid response.GS could upregulate GRs which were down regulated by GC in clinical.