Expression Of Notch1/Jagged1 On Peripheral Lymphocytes And Allograft Renal Biopsies Of Renal Transplantation Recipients

Objective and SignificanceRenal transplantation is an effective clinic method to cure end-stage renal deseases.In renal transplantation field,acute rejection(AR) is the most frequent post-operation complication.AR often took place in the first 3 months,especially 1-3 weeks after operation.AR can lead to chronic rejection(CR) and is one of the most dangerous factors to result in allograft function losing.If AR gains early diagnosis and correct treatment,90%of ARs can be reversed.Acute tubular necrosis(ATN) is also a kind of common complication after renal transplantation.Main symptom of ATN and AR are both oliguria and anuria.ATN and AR are difficultly to be identified in early stage after operation.So it is important to early diagnose AR. Blood CR is not sensitive and absence lack of specificity.Renal needle biopsy is a traumatic examination.It is a hot point of clinic research to study non-traumatic diagnosis methods to AR.Notch signal pathway is a group of transmembrane receptors.It is made up of Notch receptor,Notch ligand,CSL-DNA fixation protein,other effector and Notch regulatory molecules.Four kinds of Notch gene were found mammalian which encode four Notch receptors(Notch1～4).There are 5 kinds of Notch ligands (Delta-like1,3,4 and Jagged1,2) in mammalian.Notch ligands are transmembrane receptors too.Jagged1(CD339) is one of the main Notch ligands.Recently,some research discovered that Notch signal can effect on differentiation and accommodation of periphery immune system.When renal allograft rejection occurred,allograft stimulates immune system and then Notch receptor expression increases.Notch signal pathway promotes lymphocytes differentiation and proliferation.Activated lymphocytes excretes cytokine which promotes lymphocytes proliferation and brings out immunologic effect.Notch signal can enhance the activity of NF-kappa B through modifying CD25 expression on the surface of lymphocytes,and can also promote lymphocytes proliferation through promoting IFNγ,secretion.Activation of Notch signal pathway is ahead of the secreting of cytokine or CDs.So examination of Notch1 receptor expression is perhaps more helpful on early diagnosis of acute rejection.Materials and Methods1 15 patients suffering early acute rejection after renal transplantation(＜1 month) in 2007.1～2008.12 are arranged as acute rejection(AR) group.6 patients with post-operation acute tubular necrosis are arranged as ATN group.15 patients with normal post-operation renal function are arranged as control group.Flow cytometry is used to detect Notch1/Jagged1 expression on lymphocytes before operation,in one week after operation and in one week after AR and ATN occurrence.Notch1/Jagged1 expressions before operation,after operation and after morbidity are compared.2 8 patients suffering late acute rejection(＞1 year) in 2007.1～2008.12 are arranged as late acute rejection(LAR) group.15 patients with chronic allograft nephropathy(CAN) are arranged as CAN group.10 patients with stable and normal post-operation renal function are arranged as control group.Flow cytometry is used to detect Notch1/Jagged1 expression on lymphocytes.Notch1/Jagged1 expressions of every group are compared.3 Nephridial tissue samples collected in 2007.1～2008.12 are divided into 3 groups as follow:(1) AR group,15 samples gained through kidney puncturation when acute rejection occurred.(2) CAN group,15 samples gained from CAN patients.(3) Control group,10 samples are normal kidney tissue.The expression of Notch1 and Jagged1 were detected by immunohistochemistrical SP staining method.Notch1/ Jagged1 expressions of every group and among different pathological grades are compared.Results1 Preoperative Notch1 expression on lymphocytes among AR group,ATN group and control group didn't show difference(F＝0.277,P＝0.760).Notch1 expression among 3 groups didn't show difference on post-operative 1d,3d either(F＝0.123, P＝0.885 and F＝1.726,P＝0.104).However,Notch1 expression among 3 groups showed significant difference on post-operative 7d,14d(F＝3.354,P＝0.027 and F＝2.672,P＝0.039).Notch1 expression of AR group was higher than that of control group and ATN group(all P＜0.05).Notch1 expression of control group and that of ATN group didn't show difference on every day(all P＞0.05).Notch1 expression on post-operative 1d was higher than preoperative ones on every group(P＝0.000). Notch1 expression of control group decreased and reached preoperative level on post-operative 3d(P＝0.703).Notch1 expression of AR group decreased on post-operative 3d too,but increased on post-operative 7d,14d(P＝0.000 vs preoperative level).Notch1 expression of ATN group decreased slowly within post-operative 1 week.Notch1 expression of ATN group decreased and reached preoperative level from post-operative 3d(P＞0.05).Preoperative Jagged1 expression among AR group,ATN group and control group didn't show difference(F＝0.298,P＝0.744).Jagged1 expression among 3 groups didn't show difference on post-operative 1d,7d either(F＝0.032,P＝0.968 and F＝0.656,P＝0.526).Jagged1 expression of AR group was higher than that of control group and ATN group on post-operative 7d,14d(all P＜0.05).Jagged1 expression of control group and that of ATN group didn't show difference on every day(all P＞0.05). Jagged1 expression on post-operative 1d was higher than preoperative ones in all group(all P＝0.000).Jagged1 expression of control group decreased from post-operative 3d and reached preoperative level(P＝0.102).Jagged1 expression of AR group decreased on post-operative 3d too,but increased on post-operative 7d,14d and higher than preoperative level(P＝0.000).Notch1 expression of ATN group decreased on post-operative 3d too(P＝0.071),and reached preoperative level on post-operative 7d,14d(P＝0.057 and P＝0.534).2 Notch1 expression of AR group was significantly higher than that of ATN group on 1d after morbidity and on post-treatment 1d,3d 7d(t＝49.521,P＝0.000; t＝37.284,P＝0.000;t＝7.295,P＝0.000;t＝2.249,P＝0.037).They didn't show difference on post-treatment 14d(t＝1.741,P＝0.098).When acute rejection occurred, Notch1 expression of AR group was higher than preoperative one(P＝0.000).After anti-rejection,Notch1 expression decreased continually and reached preoperative level on 7d(P＝0.072).Notch1 expression of ATN group was higher than preoperative one(P＝0.002).After therapy.Notch1 expression decreased(P＝0.067 on 3d).Jagged1 expression of AR group was significantly higher than that of ATN group on morbidity day and post-operative 1d,3d(t＝60.598,P＝0.000;t＝44.716, P＝0.000 and t＝8.455,P＝0.000).They didn't show difference on post-operative 7d, 14d(t＝0.022,P＝0.884;t＝0.097,P＝0.613).Jagged1 expression of AR group was higher than preoperative on post-operative 1d(P＝0.000).After anti-rejection,Jagged1 expression decreased continually and reached preoperative level on 7d(P＝0.512). Jagged1 expression of ATN group was higher than preoperative on post-operative 1d(P＝0.005).After therapy.Jagged1 expression decreased(P＝0.054 on 3d).3 In late post-operative time,Notch1 expression on lymphocytes among AR group,CAN group and control group show significant difference(F＝43.992, P＝0.000).Notch1 expression of AR group was significant higher than that of CAN group and control group(both P＝0.000).Notch1 expression of CAN group show no difference with that of control group(P＝0.734).Notch1 expression of AR group decreaSed clearly after anti-rejection(t＝5.511,P＝0.001).Jagged1 expression among AR group,CAN group and control group show significant difference too(F＝61.652,P＝0.000).Jagged1 expression of AR group was significant higher than that of CAN group and control group(both P＝0.000).Jagged1 expression of CAN group show no difference with that of control group(P＝0.555). Jagged1 expression of AR group decreased clearly after anti-rejection(t＝3.983, P＝0.005).Notch1/Jagged1 expression of CAN group wasn't in relation to hemodialysis (t＝0.459,P＝0.654;t＝1.638,P＝0.125).There weren't difference between CsA and Tac immunological suppression programs(all P＞0.05).4 Notch1/Jagged1 mainly expressed in kytoplasm and membrame of renal tubular epithelial cell.Notch1/Jagged1 expression of normal renal tissues was very low(MOD＝0.012±0.003 and MOD＝0.009±0.003).Notch1/Jagged1 expression of AR renal tissues were higher than that of normal renal tissues(P＝0.000).Notch1/Jagged1 expression of CAN renal tissues were very weak too(MOD＝0.013±0.003 and MOD＝0.012±0.002).It was clearly lower than that of AR group(both P＝0.000),but showed no difference with that of normal renal tissues(P＝0.756 and P＝0.335).Notch1 expression of pathological gradeⅠof AR renal tissue(MOD＝0.126±0.013) showed no difference with that of pathological gradeⅡ(MOD＝0.136±0.011) (t＝1.541,P＝0.147).But Jagged1 expression of pathological gradeⅡ(MOD＝0.136± 0.005) was higher than that of pathological gradeⅠ(MOD＝0.123±0.008)(t＝3.745, P＝0.002).Notch1/Jagged1 expression of CAN renal tissues showed no difference among different pathological grades(F＝0.811,P＝0.467 and F＝0.811,P＝0.467).5 Notch1 and Jagged1 expression on peripheral blood lymphocytes showed significant positive correlation(r＝0.994,P＝0.000).Notch1 and Jagged1 expression on allograft tissue showed significant positive correlation too(r＝0.983,P＝0.000).Conclusions1 When post-renal-transplantation acute rejection occurred,Notch1/Jagged1 expression on lymphocytes increased significantly.Examination of Notch1/Jagged1 expression on lymphocytes is helpful to diagnosis of acute rejection.2 When acute rejection was reversed,Notch1/Jagged1 expression decreased significantly.So detection of Notch1/Jagged1 expression on lymphocytes can help understand the effect of anti-rejection.3 On the 1d after renal transplantation,Notch1/Jagged1 expression increased in all groups.Notch1/Jagged1 expression of ATN group didn't show difference with control group,but significantly lower than that of AR group,which was helpful to differentiate ATN and AR.4 Examination of Notch1/Jagged1 expression in late stage after renal transplantation can't diagnose CAN,but still was helpful to identify acute rejection and chronic rejection,and was helpful to understand anti-rejection effect.5 Notch1/Jagged1 expression of CAN patients isn't in relation to hemodialysis.6 Notch1/Jagged1 expression in acute rejection renal tissue was higher than that in CAN renal tissue,which shows that examination of Notch1/Jagged1 expression in allograft can help understand pathological change and prognosis of allograft.