Intragraft gene expression profiles by cDNA microarray in small-for-size liver grafts.
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.
Man
Kwan
K
Centre for the Study of Liver Disease, The University of Hong Kong Medical Centre, Queen Mary Hospital, China.
Lo
Chung-Mau
CM
Lee
Terence Kin-Wah
TK
Li
Xian-Liang
XL
Ng
Irene Oi-Lin
IO
Fan
Sheung-Tat
ST
D000818:Animals; D003199:Computer Systems; D016207:Cytokines / Q000097:blood / Q000235:genetics; D015536:Down-Regulation; D019332:Endothelin-1 / Q000097:blood / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics; D051547:Heme Oxygenase-1; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D016031:Liver Transplantation / Q000379:methods*; D008297:Male; D008854:Microscopy, Electron; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017466:Receptors, Endothelin / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016207:Cytokines; D019332:Endothelin-1; D017466:Receptors, Endothelin; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1