The Regulation Of Prednisone On The Expression Of HLA In Nephrotic Syndrome On Cultured Peripheral Blood Mononuclear Cells Induced By IFN-γ And The Mechanism Research

Objective:To study the regulation of glucocorticoids on the expression of HLA role from molecular and gene level by investigating the difference of HLA's expression in INS children's, normal PBMC interventied by prednisolone and the accommodation of CⅡTA,which provides theories for its clinical treatment.Methods:1.INS PBMC,after stimulation with IFN-γand PHA activation with different concentrations of prednisolone,was detected by flow cytometry INS PBMC constitutively activated and before and after prednisone,HLA-Ⅰ/Ⅱmolecule expression and the normal control group for comparison.2.Normal PBMC was stimulated with IFN-γ,PHA and activated with certain concentrations of prednisolone.The PBMC's CⅡTA mRNA was amplified by RT-PCR,analysed its transcription level in three groups-undisposed PBMC,activated PBMC and disposed PBMC.Results:1.the inhibition effect of prednisone on HLA expression.(1)INS cultured normal human PBMC and constitutively expressed HLA(the average fluorescence intensity were HLAⅠ:113.33±2.08,68.53±3.39;HLAⅡ:26.87±0.78,2.28±0.12;HLA-DR7:7.77±0.55,6.97±0.74,P values were＜0.01),INS also higher than normal (especially HLA classⅡmolecules prominent,it is 11.78 times to normal).(2)The expression of HLA than activation significantly increased by the PHA and IFN-γ(the average fluorescence intensity were HLAⅠ:187.33±0.58,158.00±17.35;HLAⅡ: 41.10±0.46,5.27±0.38;HLA-DR7:13.33±1.53,10.87±1.41.Pvalues were＜0.01)(3)By different concentrations of prednisone(10^-4M,10^-6M,10^-8M)treatment,note by prednisone significantly inhibited the expression of HLA(the average fluorescence intensity were HLAⅠ:157.33±8.62,138.67±15.53 and 159.67±2.51,142.00±3.46 and 169.33±5.29,147.00±4.72.HLAⅡ:35.43±3.72,3.48±0.19 and 36.40±5.40,3.33±0.21 and 36.47±0.24,3.22±0.67,HLA-DR7:9.45±0.38,6.43±0.76 and 9.28±0.34,6.59±0.77 and 9.51±0.41,6.40±0.43.P values were＜0.01)(4)By different concentrations of prednisone(10^-4M,10^-6M,10^-8M)after pretreatment,note by prednisone significantly inhibited the expression of HLA(the average fluorescence intensity were HLAⅠ:122.00±3.60,114.33±8.73and 125.67±2.31,121.33±8.02 and 127.33±4.93,123.67±5.77;HLAⅡ:21.17±0.40,3.57±0.28 and 23.13±1.53.3.75±0.26 and 26.10 ±0.40,3.85±0.17:HLA-DR7:7.76±0.22,7.65±1.16 and 8.39±0.54,7.53±0.24 and 8.01±0.74,7.70±0.50,Compared with the activation of both P＜0.01).Only prednisone pretreatment group HLA classⅡmolecules demonstrated a dose-dependent manner(P＜0.01)with the increased concentration of prednisone,the inhibitory effect of the stronger.2.The effect of prednisone on CⅡTA transcription:normal human PBMC constitutivelyexpressed CⅡTA,increased by the PHA and IFN-γ,prednisone significantly inhibited it.Conclusion:1.There are large number of HLAⅠ/ClassⅡmolecules and a certain level of HLA-DR7 constitutively expressed(expression than normal control)in a not activated state in INS PBMC,suggesting that children with INS abnormal activation of T cells,INS may be involved in the immune injury.2.Prednisone has inhibition on the expression of PBMC HLA that induced activation by IFN-γand this negatively regulation may be one of the mechanisms of immune suppression.3.Prednisone has inhibition on the expression of PBMC CⅡTA that induced activation by IFN-γ,the descend adjusting to the transcribe of CⅡTA may have something to do with the decrease expression of HLA.