冷/热缺血再灌注损伤对大鼠肝脏细胞能量状态、凋亡和胀亡的影响

目的研究冷/热两种条件下肝脏缺血再灌注损伤细胞能量状态与细胞死亡方式的关系,探讨减少此类损伤的措施。方法建立SD大鼠肝门阻断热缺血再灌注模型及大鼠原位肝移植冷缺血再灌注模型,测定术后缺血1h、再灌注1、12、72h肝组织能量物质变化,凋亡、胀亡和坏死细胞,观察组织病理学变化。结果热缺血期肝组织损伤重,肝脏细胞内ATP迅速下降为对照组的21%,肝脏细胞死亡方式以胀亡为主;再灌注后,ATP开始恢复,细胞死亡方式逐渐转为以凋亡为主。而冷缺血肝脏细胞缺血期肝组织损伤相对轻,ATP仅下降为对照组的68%,细胞死亡以凋亡为主;再灌注后12hATP即完全恢复,凋亡也达到顶峰,于72h逐渐下降。相关分析表明ATP、能荷与胀亡细胞数量呈显著负相关(r=-0.774,P<0.01;r=-0.789,P<0.01),与凋亡细胞数量无明显相关。结论(1)缺血再灌注发生后,以能量状态为中心环节,轻度损伤时细胞以凋亡为主,自主清除DNA受损的细胞;损伤严重时则胀亡为主,被动清除受损细胞,提示术前或术中给予能量物质可能有利于减少胀亡及炎症反应,延长肝门阻断时间。(2)单纯以凋亡或胀亡作为衡量组织功能的唯一指标值得探讨。

Effect of cold/worm ischemia/reperfusion on apoptosis,oncosis and energy state of hepatic cell in rat

Objective To study a pathway to reduce the injury of ischemia/reperfusion.Method SD rats were divided into 9 groups randomly.Rats models of warm ischemia/reperfusion,cold ischemia/reperfusion of orthotopic liver transplantation were made.The number of cells with normal,apoptosis,oncosis and necrosis,and the change in energy state of hepatocytes at 1 h of ischemia,1 h,12 h and 72 h after reperfusion were measured.At the same time,the histological change of liver was also observed.Results ATP level in warm ischemia groups was reduced immediately,and oncosis was the predominant death pattern of hepatocytes during the ischemia phase of 1 h.After reperfusion,ATP level was increased,then the death pattern of hepatocytes conversed into apoptosis.However,ATP was decreased slowly and apoptosis was the dominant death pattern of hepatocytes during the ischemia phase of 1 h in cold ischemia groups.ATP was increased to normal level after 12 h of reperfusion.At the same time,the number of apoptosis hepatocytes was increased and laid in summit,then fell off.Correlation analysis showed that there was a negative correlation among ATP,energy charge (EC) and oncosis hepatocytes (r=~-0.774, P<0.01;r=~-0.789, P<0.01).There was no significant correlation between ATP and the number of apoptosis hepatocytes.Liver inflammation was severe in warm I/R group during phrase of warm reperfusion and slight in cold I/R group.Conclusion Energy state plays an important role in process of I/R,which affects the death pattern of hapatocytes.And both oncosis and apoptosis are adaptation responses to stress of different level.Supplying energy to patients before/during operation should be useful to reduce R/I injury.