Man Kwan, Lo Chung-Mau, Lee Terence Kin-Wah, Li Xian-Liang, Ng Irene Oi-Lin, Fan Sheung-Tat
Centre for the Study of Liver Disease, The University of Hong Kong Medical Centre, Queen Mary Hospital, China.
Liver Transpl. 2003 Apr;9(4):425-32. doi: 10.1053/jlts.2003.50066.
The aim of this study is to identify the molecular mechanism of small-for-size graft injury through large-scale expression measurement of intragraft gene profile by carrier DNA (cDNA) microarray screening in liver transplantation. The studies compared 1,081 intragraft genes expression profiles using cDNA microarray of small-for-size grafts (<30% of recipient liver weight) with those of whole grafts (control group) 1, 3, and 24 hours after reperfusion in a rat liver transplantation model. Intragraft gene expression was detected by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR). Hepatic ultrastructural features were shown by electron microscopy. In the small-for-size grafts, by cDNA microarray study, the vasoconstriction genes were found up-regulated together with adhesion molecules at 1 hour after reperfusion. Three and 24 hours after reperfusion, the vasopressin genes were found up-regulated together with adhesion molecules, inflammatory mediators and cell death signals, accompanied with down-regulation of the genes related to energy metabolism. By quantitative RT-PCR, intragraft messenger RNA (mRNA) expression of endothelin-1 (ET-1) and endothelin-1 receptor A (ETA) was up-regulated during the first 24 hours after reperfusion accompanied with down-regulation of heme oxygenase-1 (HO-1). The intragraft mRNA and plasma levels of inflammatory cytokines (interleukin [IL]-6, IL-15, tumor necrosis factor [TNF]-alpha) also were overexpressed during the first 24 hours after reperfusion. Sinusoidal congestion and disruption were found accompanied with mitochondrial swelling during the first 24 hours after reperfusion. The up-regulation of intragraft vasoconstriction genes accompanied by early overexpression of adhesion molecules and apoptotic signals, as well as down-regulation of HO-1 in small-for-size grafts may be related to sinusoidal injury leading to graft damage in liver transplantation.
本研究旨在通过肝移植中载体DNA（cDNA）微阵列筛选对移植肝内基因谱进行大规模表达测定，以确定小体积移植肝损伤的分子机制。本研究在大鼠肝移植模型中，比较了小体积移植肝（<受体肝脏重量的30%）与全肝移植（对照组）在再灌注后1小时、3小时和24小时使用cDNA微阵列的1081个移植肝内基因表达谱。通过定量逆转录聚合酶链反应（RT-PCR）检测移植肝内基因表达。通过电子显微镜观察肝脏超微结构特征。在小体积移植肝中，通过cDNA微阵列研究发现，再灌注后1小时血管收缩基因与黏附分子一起上调。再灌注后3小时和24小时，发现血管加压素基因与黏附分子、炎症介质和细胞死亡信号一起上调，同时能量代谢相关基因下调。通过定量RT-PCR，发现再灌注后最初24小时内移植肝内内皮素-1（ET-1）和内皮素-1受体A（ETA）的信使RNA（mRNA）表达上调，同时血红素加氧酶-1（HO-1）下调。移植肝内炎症细胞因子（白细胞介素[IL]-6、IL-15、肿瘤坏死因子[TNF]-α）的mRNA和血浆水平在再灌注后最初24小时也过度表达。再灌注后最初24小时发现肝血窦充血和破坏，同时伴有线粒体肿胀。小体积移植肝内血管收缩基因上调，伴有黏附分子和凋亡信号早期过度表达，以及HO-1下调，可能与肝移植中导致移植肝损伤的肝血窦损伤有关。
