大鼠肝移植缺血再灌注损伤对肝细胞凋亡、胀亡及再生的影响

目的：通过对大鼠原位肝移植缺血再灌注损伤所致肝细胞两种细胞死亡方式的动态研究，结合细胞再生情况。探讨减轻此类损伤的途径．方法：SD大鼠随机分为5组，对照组(n=5)麻醉后开腹取材；其余4组(n=40，每组5对)建立大鼠同种异体原位肝移植模型，分别于术后0、1、12和72h取材．(1)流式细胞仪上机检测Annexinv与碘化丙锭混匀双染色的肝细胞，计数正常、胀亡、凋亡、坏死4种细胞百分数；(2)流式细胞仪检测G1期、S期、G2期肝细胞，计算增殖指数．结果：与对照组比较，供肝冷缺血1h后凋亡细胞显著增多，再灌注后继续增多，并于72h逐渐下降；坏死和胀亡细胞变化不大．G-2期细胞、增殖指数于再灌注后显著增加，在72h仍处于高峰．结论：肝移植过程中细胞凋亡是肝细胞死亡的主要形式，肝移植后存在细胞增殖，增强肝细胞的再生能力也是减轻损伤的一种途径．再灌注期凋亡的作用和机制均值得进一步研究．

Effect of Ischemia-reperfusion Injury on Apoptosis,Oncosis and Regeneration of Hepatocytes in Rat

Objective: In order to approach a pathway to reduce ischemia-reperfusion injury, we studied the relationship of apoptosis, oncosis and cell proliferation induced by ischemia -reperfusion after liver transplantation.Methods:SD rats were divided into 5 groups randomly.Rats ( n =5) in control group were performed anesthesia and sham operation.Liver transplantation models ( n =40,5 pairs of each group ) were established in other four groups.Liver tissue was taken at 0, 1,12 and 72h after operation.The amounts of hepatocytes including normal, apoptosis, oncosis and necrosis dyed with a double staining method of Annexin V and propidiom iodide (PI) were measured by bivariate flow cytometry(FCM).Hepatocytes in phase of G1, S and G2 were detected by flow cytometry.At the same time, proliferation indices were also counted.Results: The number of apoptosis hepatocytes increased significantly 1h following cold ischemia and continued to increase during the period of reperfusion.Then gradually decreased 72h after liver transplantation.The cells in phase of G2 and the proliferation indices increased significantly after reperfusion, and laid in summit at 72h after liver transplantation.Conclusion:Cell apoptosis is a primary death model of hepatocyte in liver transplantation.After liver transplantation, cell proliferation occurs.Enhanced capacity of hepatocyte regeneration can relieve ischemia-reperfusion injury.The effect and mechanism of apoptosis during reperfusion deserves further research.