The experimental study about liver grafts from Marginal Donor

背景。为了成功进行非心脏供体肝移植，有必要明确肝移植损伤的机制。本研究探讨了预冷保存灌注对移植物能量状态和微循环维持的影响。我们还研究了抗氧化剂胆绿素和自由基清除剂依达拉奉对肝移植物炎症反应的影响。使用雄性Wistar大鼠。取出肝脏，在UW溶液中保存，用含氧Krebs-Henseleit溶液灌注60分钟。将大鼠分为以下6组（n=5）： i)对照组；无热缺血（WI）和冷保存，ii)HBD组；无WI冷保存6小时，iii)NHBD组；WI和冷保存30min， iv)PRE组；与WI，心脏骤停后30分钟预冷保存灌注，冷保存，v)BV组；预冷保存灌注中加入胆绿素；vi)ED组；胆绿素在预冷保存灌注中的添加。观察再灌注时门静脉血流和胆汁生成、灌注液中AST和TNF-α、组织中能量电荷（EC）和ATP水平及组织学变化。PRE、ED、BV组门静脉流量均高于NHBD组。PRE、ED和BV组的胆汁产量也高于NHBD组。BV组AST和TNF-a的升高明显降低。BV组再灌注后EC和ATP水平高于NHBD组。预冷保存灌注和加入依达拉奉或胆绿素灌注可提高NHBD移植物的存活率。结果表明，保持移植物的能量状态和微循环是成功移植NHBD的重要因素。

Background. For a successful liver transplantation (LTx) from non-heart-beating donors (NHBD), it is necessary to clarify the mechanism of the liver grafts injuries. In this study we investigated the effects of pre-cold preservation perfusion on the grafts for the purpose of the maintenance of the energy status and microcirculation. We also investigated the effects of the antioxidant, biliverdin, and free radical scavenger, edaravone on liver grafts to reduce inflammatory reaction.Methods. Male Wistar rats were used. Livers were retrieved, preserved in UW solution, and perfused for ) 60 minutes with oxygenated Krebs-Henseleit solution. Rats were allocated to six groups as follows (n=5) : i)Control group; no warm ischemia (WI) and cold preservation, ii)HBD group; no WI with cold preservation for 6 hours, iii)NHBD group; with 30 minutes of WI and cold preservation, iv)PRE group ; with WI, 30 minutes pre-cold preservation perfusion after cardiac arrest, and cold preservation, v)BV group; addition of biliverdin to pre-cold preservation perfusion, vi)ED group; addition of biliverdin to pre-cold preservation perfusion. Portal flow and bile production during reperfusion, AST and TNF-α in perfusate, energy charge (EC) and ATP level in the tissue, and histological findings were investigated.Results. Portal flow volume in the PRE, ED, and BV groups were higher than in NHBD group. Bile production in the PRE, ED, and BV groups were also higher than in the NHBD group. Increase of AST and TNF-a was reduced in the BV group. EC and ATP level of the BV group after reperfusion were higher than those of the NHBD group.Conclusions. Pre-cold preservation perfusion and addition of edaravone or biliverdin to perfusate improved viability of grafts from NHBD. The results mean that to preserve the energy status and microcirculation of the graft is important for successful LTx from NHBD.