冷保存时间对肝脏移植大鼠供肝线粒体ATPase6、ATPase8基因表达的影响及意义

Effect of different cold preservation time on ATPase6,ATPase8 expression in rats liver transplantation

目的探讨冷保存时间对移植肝脏能量代谢的影响及机制。方法取Wistar大鼠186只,其中180只采用改良"二袖套法"制作大鼠肝移植模型并分为A、B、C组各60只,供肝冷保存时间分别为30 min、6 h、12 h;另6只为D组,行假手术对照。分别于制模后12 h、24 h、3 d、5 d、7 d采集肝组织标本检测各组肝脏线粒体AT-Pase6、ATPase8 mRNA表达及ATP含量。结果与D组比较,术后12 h A、B、C三组ATPase6、ATPase8 mRNA表达及ATP含量均降低,且C组显著低于A、B组(P<0.05),且ATPase6、ATPase8 mRNA变化趋势一致;24 h后A、B、C组ATPase6、ATPase8 mRNA表达及ATP含量开始升高,C组ATPase6、ATPase8 mRNA表达升高较A、B组缓慢。结论肝脏移植过程中,供肝冷保存时间延长可导致能量合成障碍,机制可能为下调ATPase6、ATPase8基因表达。

Objective To study the effect of cold preservation time on the expression of mitochondria（mt） DNA encoding ATPase6,ATPase8 of hepatocytes in rat liver transplantation. Methods Orthotopic liver transplantations were performed in wistar rats using the cuff technique as described by Kamada with modifications. Rats were randomly divided into four groups namely：group A （cold preservation of 30 rain）, group B （cold preservation of 6 hours）, group C （cold preservation of 12 hours） and group D （sham operation）. Samples of Liver were collected at 12 h,24 h and on 3 rd, 5 th, 7 th days after operation. The ATP of liver were observed in each group, mtDNA encoding ATPase6, ATPase8 mRNA were determined by RT-PCR. Results The expression of ATPase6,ATPase8 mRNA decreased in groups A, B and C at 12 h after operation. ATPase6,ATPase8 mRNA in group C were the lowest compared with groups A and B （P 〈 0.05 ）. The expres- sion of ATPase6, ATPase8 mRNA and ATP increased 24 h later in groups A, B and C. Both ATPase6 and ATPase8 mRNA in group C increased slowly compared with that in groups A and B 24 h later. Conclusions Cold preservation time delay of donor liver can lead to disorder of ATP metabolism, the mechanism may downregulate the gene expression of ATPase6 and ATPaseS.