Comprehensive genome analysis using DNA microarray during liver regeneration after partial hepatectomy and living donor liver transplantation

背景小肝再生过程中高胆红素血症引起的肝损害是扩大成人活体肝移植适应证的主要障碍。我们以前报道过，在大鼠90%肝切除后，多药耐药蛋白（MRP）2和MRP 3的表达在肝再生过程中的减少导致结合型高胆红素血症。然而，这些ATP结合盒（ABC）转运蛋白在肝再生过程中的基因表达谱的全面审查的遗传控制尚未进行。在90%肝切除术后0、24、72和168小时的每个时间点，从三个大鼠肝脏制备RNA。通过大鼠基因组230 2.0阵列分析基因表达谱，特别参考ABC转运蛋白。同时分析了70%肝切除术后用脂多糖处理的大鼠肝脏。在31，042个探针中，1587个报告基因被鉴定为上调或下调，超过2倍。在16个ABC转运蛋白基因中，MRP 2和OATP 1表达显著下调，而MRP 1和MRP 3有表达趋势。这些遗传变化通过实时PCR证实。在肝再生过程中加入脂多糖可协同增强这些变化。微阵列分析不仅显示了再生肝脏中广泛的基因表达谱，同时还显示了与胆红素转运相关的更特异的分子事件。ABC转运蛋白表达模式的改变，尤其是MRP 2的下调，似乎是肝再生过程中肝衰竭的关键事件。

Background. Liver damage with hyperbilirubinemia during regeneration of the small liver is the major hurdle to expand indications of adult living donor liver transplantation. We previously reported that decrease in multidrug resistance protein (MRP) 2 and expression of MRP3 during liver regeneration lead to conjugated hyperbilirubinemia after 90% hepatectomy in rats. However, the genetic control of these ATP binding cassette (ABC) transporters under comprehensive review of the gene expression profile during liver regeneration have not been performed yet.Methods. RNAs were prepared from three rat livers at each time point of 0,24,72, and 168 hr after 90% hepatectomy. Gene expression profile was analyzed by the Rat Genome 230 2.0 array with special references to ABC transporters. Those from rat livers treated with lipopolysaccharide after 70% hepatectomy were also analyzed.Results. Among 31,042 probes, 1587 reported genes were identified either up-or down-regulated, more than 2-fold. Among 16 ABC transporter genes, MRP2 and OATP1 were significantly down-regulated, while MRP1 and MRP3 tended to be expressed. These genetic changes were confirmed by real-time PCR. These changes were enhanced synergistically by adding lipopolysaccharide during liver regeneration.Conclusions. Microarray analysis demonstrated not only extensive gene expression profile in the regenerating liver but more specific molecular events related to bilirubin transport at the same time. Changes in expression pattern of ABC transporters, especially down-regulation of MRP2,seem the key event in liver failure during liver regeneration.