Proteome Analysis Of Renal Allograft Rejection In Rats

Objeetive: To analysis the alteration of protein expression patternsin rat allotransplantations transplant kidney glomerulus byproteomics technique, search and identify differential expressionproteins as biomarkers which are related with acute or chronicrejection.Methods:(1) We used Fisher rats as donors and Lewis rats asrecipients to establish the model of renal transplantation. All transplant ratswere divided into three gruops: acute rejection (1week ofpost-transplatation), chronic rejection (12weeks of post-transplatation) andnormal control. Allograft specimens were obtained at a predetermined time.(2) The total proteins of rat transplant renal tissues and normal renal tissuewere separated by menas of two-dimensional gel electrophoersis(2-DE).The diefferntial expression proteins were analyzed and thenidentified by matrix-assisted laser desoprtion/ionization time of flight massspectrometry (MALDI-TOF-MS).The function of differential expressionproteins and the relation to the renal rejection is analzyed. Resulst:(1) A total of125renal orthotopic renal transplantationoperations were completed. In late, the success rate of the operation was86.0%. The average operation time was (130±16) min. The average time ofwarm ischemia and cold ischemia was less30s and50min respectively.(2)Morphological observation: control group organization structure wasbasically normal, no inflammatory cells, hemorrhage or necrosis, interstitialedema, renal tubular epithelial cells with mild swelling. Acute rejectiongroup (post-transplantation1week): inflammatory cell infiltratied in renalinterstitial, infiltration of inflammatory cells including lymphocytes andmonocytes, but predominated by lymphocytes, which were accumulatingaround the renal tubules and capillaries. Renal tubular epithelial cellshowed swelling, necrosis, nuclear pyknosis, and arterial fibrinoid necrosis.Chronic rejection group (post-transplantation12weeks): it showedextensive interstitial fibroblast proliferation, glomerular basementmembrane thickening, hardening, occlusion, tubular atrophy degeneration.(3)The result showed that well-ersolved and reporudcible2-DE patternswere obatined. The averages post of acute rejection group, chronic rejectiongroup and control group were554±34,583±42and542±21respectively.(4) Compared to control group,37dieffrentially expressed porteins werescreened in acute rejection group, and22dieffrentially expressed porteinsin chronic rejection group.(5) The total37differential expression proteinsof acute rejection group were identified by PMF, and6differential proteins were founded by bioinformatic. In chronic rejection group,4differentialproteins were finally founded by bioinformatic.(6) It was identified thatthe expression of PDIA3was increased significantly in acute rejectiongroup as compared with that in control group by Immunohistochemistry,ELISA, RT-PCR and Western blot.Conclusion:(1) Six differential expression proteins which are maybecorrelated with acute rejection of renal allograft were identified byproteome analysis.(2) PDIA3may be involved in the developing processof the acute renal rejection, and it is a possible biological marker.(3) Fourdifferential expression proteins which are maybe correlated with chronicrejection of renal allograft were identified by proteome analysis.