Research Of Overexpression And Clinical Significance Of Lymphocyte P-gp In Transplant Recipient

Part one The expression level of lymphocytes P-gp in healthy populationObjective:To investigate the P-gp expression level of lymphoctes in healty populatioon,in order to establish the diagnosis standards in overexpression cases.Methods:Collecting lymphoctye samples from 63 healthy cases,to analyze the expression level of P-gp using RT-PCR and FACScan technique,comparising the expression level among groups classified as sex,age,lymphocyte subtypes.Results:The average epression level of lymphocyte P-gpin healthy person is 12.5%.the diffrences among the sex groups and lymphocytye subtype groups are not distinct respcetively;no diffrence among age groups except the significant lower expression in＜14yr group;the results of RT-PCR is similar to that of FACScan.Conclusions:Physically,P-gp is positively experssed in lymphocytes with a 12.5%average level.No difference are found among groups classified as sex,lymphocyte subtype,age,except the＜4yr group expressing lower P-gp. Part two Alteration of P-gp expression in transplant recipientsObjective:To investigate the alteration of lymphocyte P-gp expression in transplant recipients.Methods:Collecting the lymphocyte samples from 78 recipients in 1st,3rd,6th,12th month respectively,analyze the expression level of P-gp,and comparise the difference among groups classified as sex,age,lymphocyte subtype.Results:Averagelly,the expression level of lymphocyte P-gp in transplant recipients increase initially at 1st month,to the peak level around 3rd month,after then,keep stable at this level.No difference is between sex groups and age groups;the P-gp expression level in CD+4T cell is significantlly higher than in B cell and CD+8T cell.Conclusions:Medication of immunosuppressant may induce the overexpression of lymphocyte P-gp in transplant recipients. Part three Analysis of the induction factors of P-gp overexpressionObjevtive:To make sure the drugs responsible for the induction of P-gp overexpression in transplant recipients.Methods:Analyzing the expression level of lymphocytes P-gp in transplant recipients medicated by different regimens respectively,to eluciate the role of particular immunosuppressant in the induction of P-gp expression.Results:The expression levels of P-gp are higher and stable in recipients medicated by calcineurin inhibitors with or without prednison, meaning that glycucorticoids play a minor role in the induction of P-gp even though it is a kind of P-gp substrates.Conclusions:Calcineurin inhibitors are major factors implicated in the induction of overexpression of lymphocyte P-gp in transplant recipients. Part four Infuence of P-gp overexpression in lymphocytes on the intracellular drug accumulationObjective:To evaluate the efflux of drug from lymphocytes by the overexpressing P-gp.Methods:Compare the accumulation of Rhodamine 123 in lymphocytes with different expression level of P-gp.Results:The fluorence intensity of Rhodamine123 in lymphocytes is reversly related to the expression level of P-gp.The accumulation of Rhodamine123 are significantly less in midlle and high groups than control and low groups.Conclusions:The overexpression level of P-gp increase the drug-efflux activity from lymphocytes and decrease the intracellular concentration of therapeutic agents. Part five Inhibition of P-gp overexpression in lymphocytes on CsA pharmacological effectsObjective:To evaluate the immunosuppression efficacy of CsA under the context of alteration of lymphocytes P-gp expression in transplant recipients.Methods:using lymphocyte culture experimental system,to analyze the influence of P-gp expression level on CsA efficacy to suppress the lymphocyte proliferation and cytokine secretion by PHA application.Results:The inhibition effect of CsA on PHA-stimulusing prolieration and cytokine secretion of lymphocytes is markdly decreased in lymphocytes with middle and high P-gp level.Conclusions:The overexpression of P-gp in lymphocytes decreases immunosuppression efficacy of CsA. Part six Relation between P-gp overexpression and graft functionObjective:Analyzing the graft functions in recipients with different P-gp expression level,to investigate the relation between P-gp overexpression and graft function.Methods:78 recipients being followed up for one year,the graft function is evaluated by laboratory test including serum Crea,UN;graft hemodynamic criteria;edema,oligouriea.Each of the single alteration and occurrence is recorded as one ill-episode.Results:The sum of ill-episode during follow-up is 291.the incidence of ill-episode are significant higher in middle and high P-GP level groups.Conclusion:The level of lymphocyte P-gp is reversely related to the graft function,and can be used as a mark of diagnosis and prognosis for graft dysfunction.