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

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

Study on hepatocellular apoptosis and its relationsh ip to cellular Bax gene expression during cold preservation-reperfusion of is ol ated livers

Objective　To study the hepatocellular apo ptosis during cold preservation- reperfusion of isolated livers and its mechani s m during clinical liver transplantation. Methods　Hepatoce llular apoptosis of 4 groups of rabbit livers, which had experienced cold storag e for 0(group A), 3(group B), 6(group C), 9(group D)h respectively, were observe d during their cold preservation-reperfusion by using TdT-mediated dUTP-bioti on nick end labeling method and under an electron microscope. Simultaneously, level s of cellular Bax gene protein expression were measured in the time points of bot h pre-reperfusion and post-reperfusion. Results　Apoptot ic hepatic parenchymal cells (HPCs) were obviously found in reperfusing livers sub sequent 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, g ro up 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 al l the groups at different degrees. It was found by quantitative analysis that th ere 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.