左旋精氨酸对无心跳大鼠供肝缺血再灌注损伤防护作用研究

Protective effects of L-Arginine on non-heart-beating rat liver graft aginst ischemia-reperfusion injury

目的 :探讨左旋精氨酸 (L Arg)对无心跳大鼠供肝缺血再灌注损伤的防护作用。方法 :Wistar大鼠 1 0 4只 ,随机取 8只为正常对照组 (N组 ) ,另 96只随机分为供、受体组 ,两两随机配对后再分为 6组。C1 、C2 、C3 组为实验对照组 ,E1 、E2 、E3 组为实验组。C1 与E1 、C2 与E2 、C3 与E3 分别行活体供肝肝移植和心跳停搏后 30min、4 5min供肝肝移植 ;N组仅于开腹后抽取下腔静脉血及切取肝组织备检。其中各实验组供肝保存液中加入L Arg(1mmol/L) ,受体鼠在移植后门静脉 (PV)复流前 1 0min从尾静脉注射L Arg(1 0 0mg/kg) ;各实验对照组则用等量生理盐水作对照。移植完成后 ,于PV复流后 1h、3h、2 4h检测血清一氧化氮 (NO)、丙氨酸氨基转移酶 (ALT)、天冬氨酸氨基转移酶 (AST)及血浆内皮素 (ET)水平 ,观察肝组织超微结构。结果 :与N组相比 ,各实验组与实验对照组大鼠PV复流后 1h血清NO水平明显降低 (P <0 .0 5 ) ,血浆ET水平、血清ALT、AST水平明显升高 (P <0 .0 5 ) ;与对应实验对照组相比 ,各实验组在PV复流后 1h、3h、2 4h血清NO水平明显升高 (P <0 .0 5 ) ,血清ALT、AST、血浆ET水平明显降低(P <0 .0 5 ) ;各实验对照组之间相比 ,C3 、C2 组血清NO水平明显低于C1 组 (P <0 .0 5 ) ,C3 组明显低于C2 组

Aim:To study the protective effects of L Arginine(L Arg) against ischemia reperfusion liver injury of non heart beating donors(NHBO) in rat liver graft. Methods: A total of 104 Wistar rats were randomly divided into 7 groups: normal control group ( n =8),control 1, 2, and 3 group(C 1, C 2, C 3, n =16), experimental group 1, 2,and 3(E 1, E 2, E 3, n =16). For group C 1 and E 1 , group C 2 and E 2 , group C 3 and E 3 , the warm ischemic time was 0, 30, and 45 min,respectively. Liver grafts were flushed with and preserved in 4℃ Euro collins solution containing 1 mmol/L L Arg for 1 h in each experimental group. Recipients of each experimental group were injected with L Arg (10 mg/kg body weight) by tail vein 10 min before portal vein reperfusion. Donors and recipients of each experimental control group were treated with normal saline. Then transplantation was performed. At 1, 3, and 24 h after portal vein reperfusion, blood samples were obtained to determine the levels of ALT, AST, NO 2 -/NO 3 -, and ET. At 3 h after portal vein reperfusion, grafts samples were fixed by 2.5% glutaradehyde for electronscopy observation. Results:At 1 h after portal vein reperfusion, the levels of NO 2 - /NO 3 - in group E 1, E 2, E 3 and group C 1, C 2, C 3 were significantly lower while plasm ET,serum ALT and AST were significantly higher than that in normal control group( P <0.05); at 1 h, 3 h, 24 h, the levels of NO 2 -/NO 3 - in group E 1,E 2,E 3were significantly higher while plasm ET,serum ALT and AST were significantly lower than in corresponding control groups (C 1, C 2, C 3)( P <0.05). The levels of NO 2 -/NO 3 - in group C 2 and C 3 was significantly lower than in group C 1 ( P <0.05), and that in group C 3 was significantly lower than in group C 2 ( P <0.05).At 1 h, 3 h, and 24 h, the levels of plasm ET, serum ALT,and AST in groups E 1, E 2, E 3 were significantly lower than in corresponding control groups(C 1, C 2, C 3)( P <0.05). The levels of plasm ET,serum ALT, and AST in group C 3 were significantly lower than in group C 1 ( P <0.05), and the levels of plasm ET,serum ALT,and AST in group C 3 were significantly lower than in group C 2 ( P < 0.05). Pathological changes in group E 1, E 2, E 3 were significant milder than in corresponding experimental control groups(C 1, C 2, C 3). Conclusions:The unbalance between NO and ET plays an important role in the development of ischemia reperfusion injury of liver graft in NHBD. L Arg could attenuate liver graft injury in NHBD by improving the balance between NO and ET.