lznerowicz A, Chudoba P, Kamińska D, Kościelska-Kasprzak K, Drulis-Fajdasz D, Hałoń A, Janczak D, Boratyńska M, Klinger M, Patrzałek D, Polak W G
Department of Vascular, General and Transplant Surgery, Wroclaw Medical University, Wroclaw, Poland.
Transplant Proc. 2011 Oct;43(8):2887-90. doi: 10.1016/j.transproceed.2011.08.013.
Apoptosis is one of the most important mechanisms leading to kidney graft injury during transplantation. The aim of this study was to assess the expression of genes involved in apoptosis in transplanted kidneys derived from deceased donors (DD) at various stages of the transplant procedure, seventy eight transplanted kidneys procured from 43 DD were included in this study. As a baseline control for gene expressions we used six kidney allografts obtained from living donors (LD). Three core biopsies were performed: biopsy 1--5 minutes before organ perfusion in the donor; biopsy 2--at the end of cold ischemia before kidney implantation; and biopsy 3--30 minutes after reperfusion. Tumor protein p53 (TP53), caspase-3 (CASP3), B-cell lymphoma 2 protein (Bcl2), and heme oxygenase 1 (HO-1) gene expression levels were determined using custom-designed low-density arrays (TaqMan assay). Comparison of gene expression between DD and LD kidneys revealed greater expression of all genes in kidneys from DD in all biopsies; however, only CASP3 expression in biopsy 1 and TP53 expression in biopsy 3 were statistically significant. Prolongation duration of brain death beyond 10 hours in DD resulted in a significantly decreased CASP3 expression in biopsy 1. When the cold ischemia time (CIT) was longer than 24 hours, the expressions of Bcl2, TP53, and CASP3 were significantly higher compared to kidneys with ClT<24 hours. There was no correlation between warm ischemia time and gene expression in biopsy 3. CASP3 and TP53 expression only in biopsy 1 were significantly higher among kidney allografts with delayed (DGF) compared with immediate graft function. In conclusion expression of genes involved in apoptosis was more pronounced in kidney allografts from deceased donors. A prolonged donor brain-death period beyond 10 hours resulted in decreased CASP3 expression. CIT longer than 24 hours was associated with increased expressions of Bcl2, TP53, and CASP3. CASP3 and TP53 expressions were significantly higher among kidneys allografts displaying DGF.
细胞凋亡是移植过程中导致肾移植损伤的最重要机制之一。本研究的目的是评估在移植过程不同阶段来自脑死亡供者（DD）的移植肾中参与细胞凋亡的基因表达情况。本研究纳入了从43例脑死亡供者获取的78个移植肾。作为基因表达的基线对照，我们使用了从活体供者（LD）获得的6个同种异体肾移植。进行了三次核心活检：活检1——在供者器官灌注前5分钟；活检2——在肾脏植入前冷缺血结束时；活检3——再灌注后30分钟。使用定制设计的低密度阵列（TaqMan分析）测定肿瘤蛋白p53（TP53）、半胱天冬酶-3（CASP3）、B细胞淋巴瘤2蛋白（Bcl2）和血红素加氧酶1（HO-1）基因的表达水平。DD肾和LD肾之间的基因表达比较显示，在所有活检中，DD肾中所有基因的表达均更高；然而，仅活检1中的CASP3表达和活检3中的TP53表达具有统计学意义。脑死亡供者脑死亡持续时间超过10小时导致活检1中CASP3表达显著降低。当冷缺血时间（CIT）长于24小时时，与CIT<24小时的肾脏相比，Bcl2、TP53和CASP3的表达显著更高。活检3中热缺血时间与基因表达之间无相关性。与立即具有移植肾功能的同种异体肾相比，延迟移植肾功能（DGF）的同种异体肾中仅活检1中的CASP3和TP53表达显著更高。总之，参与细胞凋亡的基因在脑死亡供者的同种异体肾移植中表达更为明显。供者脑死亡持续时间超过10小时导致CASP3表达降低。CIT长于24小时与Bcl2、TP53和CASP3表达增加有关。在显示DGF的同种异体肾中，CASP3和TP53表达显著更高。
