肝脏低温保存-再灌注期间肝细胞凋亡及其与Bax基因蛋白相关性的研究  

Study on hepatocellular apoptosis and its relationship to cellular Bax gene expression during cold preservation-reperfusion of isolated livers

作　　者：汤礼军[1] 田伏洲[1] 王雨[1] 胡建中[1] 李晓军[1] 

机构地区：[1]成都军区总医院全军普通外科中心实验室,610083

出　　处：《中华实验外科杂志》2001年第2期144-146,共3页Chinese Journal of Experimental Surgery

摘　　要：目的　研究临床肝移植过程中供肝低温保存 再灌注期间肝细胞凋亡及其相关机制。方法　应用细胞凋亡原位末端标记法并结合电镜观察 ,检测低温保存时间分别为 0 (A组 )、3 (B组 )、6(C组 )、9(D组 )h的 4组兔肝脏在低温保存 再灌注过程中肝细胞凋亡情况。同时 ,对各组肝脏再灌注前后肝细胞内Bax基因蛋白表达进行测定。结果　A、B、C、D各组肝脏于低温保存后的再灌注 60min时 ,组织内可见明显的肝实质细胞凋亡 ;其凋亡细胞数量 (1× 10 -2 个 / μm2 )分别为 :1.2 1± 0 .84、4.3 8± 1.14、7.2 4± 2 .3 9、11.3 9± 3 .3 6;且 4组间差异均有显著性 (P <0 .0 5或P <0 .0 1)。此外 ,4组肝脏组织内均有程度不等的Bax蛋白表达 ,阳性物质主要分布于肝实质细胞浆内 ;定量结果显示 ,4组Bax阳性细胞数及阳性物质积分吸光度值依次为 :1.18± 0 .84、73 7.0 4±45 .5 7,3 .2 1± 1.3 0、10 0 6.5 8± 70 .10 ,6.5 6± 1.14、14 0 1.16± 10 6.93 ,11.18± 1.92、1797.2 3±13 0 .73 ;各组间差异均有显著性 (P <0 .0 5或P <0 .0 1)。结论　肝实质细胞凋亡明显参与了肝脏低温保存 再灌注损伤过程 。Objective To study the hepatocellular apoptosis during cold preservation reperfusion of isolated livers and its mechanism during clinical liver transplantation. Methods Hepatocellular apoptosis of 4 groups of rabbit livers, which had experienced cold storage for 0(group A), 3(group B), 6(group C), 9(group D)h respectively, were observed during their cold preservation reperfusion by using TdT mediated dUTP biotion nick end labeling method and under an electron microscope. Simultaneously, levels of cellular Bax gene protein expression were measured in the time points of both pre reperfusion and post reperfusion. Results Apoptotic hepatic parenchymal cells (HPCs) were obviously found in reperfusing livers subsequent to cold storage, and there was significant difference in the numbers of apoptotic HPCs among all the groups (group A: 1.21±0.84, group B: 4.38±1.14, group C: 7.24±2.39, group D: 11.39±3.36, P <0.05 or P <0.01). In addition, Bax positive expression, which was observed in cytoplasm of HPCs, appeared in all the groups at different degrees. It was found by quantitative analysis that there was significant difference in the numbers of apoptotic HPCs or the intensity of bax expression in HPCs (group A: 1.18±0.84, 737.04±45.57; group B: 3.21±1.30, 1 006.58±70.10; group C: 6.56±1.14, 1 401.16±106.93; group D: 11.18±1.92, 1 797.23±130.73, respectively, P <0.05 or P <0.01). Conclusion Apoptosis of HPCs is markedly involved in donor liver cold preservation reperfusion injury which may be related with cellular Bax gene protein.

关 键 词：兔 肝脏 低温保存 再灌注损伤 脱噬作用 BAX基因 细胞凋亡 

分 类 号：R657.3[医药卫生—外科学]