线粒体ATP酶6、8基因及编码蛋白表达改变对移植肝脏能量代谢的影响  

Altered expression of mitochondrial DNA encoding ATP 6, 8 and encoding protein in energy metabolism during cold preservation and reperfusion injury in rat liver transplantation

作　　者：张莹[1] 别平[2] 石承先[1] 

机构地区：[1]贵州省人民医院肝胆外科,贵阳550002 [2]第三军医大学西南医院全军肝胆外科研究所

出　　处：《中华医学杂志》2010年第10期708-711,共4页National Medical Journal of China

基　　金：贵州省优秀科技教育人才省长基金（2005-229）

摘　　要：目的探讨肝细胞线粒体DNAATP酶6、ATP酶8基因及编码蛋白F0F1-ATP酶表达改变对移植肝能量代谢的影响。方法采用改良“二袖套法”制作大鼠肝移植模型，随机分为A组（冷保存30min组），B组（冷保存6h组），C组（冷保存12h组）和D组（假手术对照组），分别于制模后于12h、24h、3d、5d、7d采集标本，检测各组大鼠肝脏ATP含量、mtDNAATP酶6、ATP酶8mRNA和F0F1-ATP酶蛋白表达。结果冷保存再灌注早期（12h），各组ATP酶6、ATP酶8mRNA、F0F1—ATP酶蛋白表达（ng／mg蛋白）和ATP含量（U／L）均降低，C组（0．81±0．08，0．71±0．07，0．47±0．07，0．44±0．05）显著低于A组（1．11±0．04，1．07±0．07，0．86±0．15，0．70±0．11）、B组（1．02±0．07，0．90±0．10，0．65±0．07，0．55±0．08）（P〈0．05）。再灌注24h后各组A11P酶6、ATP酶8mRNA、F0F1-ATP酶蛋白表达和ATP含量增加，C组升高较A组、B组缓慢。结论ATP酶6、ATP酶8基因表达改变后F0F1-ATPase蛋白表达异常，可能是导致能量合成障碍的主要原因。Objective To study the expression changes of mitochondrial （mt） DNA encoding ATP6, 8 of hepatocytes in rat liver transplantation with cold preservation-reperfusion and explore the relationship between the expression of F0F1-ATPase induced by changes of ATP 6, 8 genes. Methods Orthotopic liver transplantation was performed in Wistar rats using the cuff technique as described by Kamada with modifications. Rats were randomly divided into four groups： group A （30 min cold preservation）, group B （6 h cold preservation）, group C （12 h cold preservation） and group D （sham operation）. Samples were collected at 12 h, 24 h and on Day 3, 5 and 7 post-operation. The levels of mtDNA encoding ATPase 6, ATPase 8 mRNA were determined by RT-PCR. The expression changes of F0F1-ATPase protein were examined by Western blot. The ATP level of liver was observed in each group. Results The expression of ATPase 6, ATPase8 mRNA, F0F1-ATPase protein expression （ ng/mg protein） and the level of ATP decreased （U/L） in groups A, B and C at 12 h post-operation. The lowest was in group C （0. 81 ±0. 08, 0. 71± 0. 07, 0. 47 ± 0. 07, 0. 44 ± 0. 05 ） when compared with groups A （ 1.11 ± 0. 04, 1.07 ± 0. 07, 0. 86 ± 0. 15, 0.70 ±0. 11） and B（1.02 ±0.07, 0.90 ±0. 10, 0.65 ±0.07, 0.55±0.08） （P 〈0.05）. The expression of ATPase 6, ATPase 8 mRNA increased after 24 h in groups A, B and C. The changes of F0F1- ATPase protein expression and ATP level were consistent with the expression of ATPase 6, ATPase 8 mRNA. Both ATPase 6 mRNA, ATPase 8 mRNA and FoF1-ATPase protein in group C increased slowly compared with groups A and B after 24 h. The mtDNA encoding ATPase 6, ATPase 8 changes of hepatocytes in rats liver transplantation with cold preservation-reperfusion suggested that mitochondrial function disorders are related to the changes of mitochondrial DNA encoding ATPase6, ATPase8. mtDNA encoding ATPase 6, ATPase 8 genes of hepatocytes decreased significantly and FoFI-ATPase protein expression decr

关 键 词：肝移植 能量代谢 基因表达 DNA 线粒体 低温保藏 

分 类 号：R657.5[医药卫生—外科学]