Quintana Alejandra B, Rodriguez Joaquin V, Scandizzi Angel L, Guibert Edgardo E
Morfologia, Departamento de Ciencias Biologicas, Facultad de Ciencias Bioquímicas, y Farmacéuticas, UNR. Suipacha 531 (S2002LRK), Rosario, Argentina.
Ann Hepatol. 2003 Apr-Jun;2(2):84-91.
Cold liver preservation in the University of Wisconsin solution (UW) followed by reperfusion alters hepatic parenchyma and extra cellular matrix. In this study we analyzed the benefit of adding either 500 microM Sodium Nitroprusside (NPNa) or 100 microM S-nitrosoglutathione (GSNO) as Nitric Oxide (NO) donors to the UW solution to prevent hepatic injury. Wistar adult rat livers were stored in UW solution (0 degrees C) for 48Hs and reperfused (60 minutes) in the isolated perfused rat liver model (IPRL). Untreated livers were used as normal controls. Livers perfused but not preserved were used as controls of reperfusion. Parenchyma damages were evaluated by Hematoxylin-Eosin stain. Picrosirius Red and Gordon-Sweets stains were used for collagen and reticulin networks, respectively. An inmunohistochemistry assay for albumin was used as functional test. Cold preservation step was followed by swollen hepatocytes with "light empty halos" surrounding the nucleus, conserved hepatocyte cords and many rounded endothelial cells. The addition of NPNa or GSNO into UW solution, avoid these alterations. Livers preserved for 48 Hs and then reperfused showed extended areas of vacuolation around central veins, and many endothelial cells were rounded and located inside sinusoidal lumens. The collagen network was disorganized while the reticulin one was less altered. Albumin was distributed preferentially in pericentral areas. On the contrary, livers preserved in presence of NPNa or GSNO did not show vacuolation and both collagen and reticulin networks were unchanged. Albumin was more homogeneously distributed in both groups. In conclusion, the addition of 500 microM NPNa or 100 microM GSNO as a NO donor, improves UW solution properties to preserve rat livers by maintaining the hepatic morphology and avoiding hepatic injury post-cold preservation/reperfusion.
在威斯康星大学溶液（UW）中进行冷肝保存，随后再灌注会改变肝实质和细胞外基质。在本研究中，我们分析了向UW溶液中添加500微摩尔硝普钠（NPNa）或100微摩尔S-亚硝基谷胱甘肽（GSNO）作为一氧化氮（NO）供体以预防肝损伤的益处。将成年Wistar大鼠肝脏在UW溶液（0℃）中保存48小时，并在离体灌注大鼠肝脏模型（IPRL）中进行再灌注（60分钟）。未处理的肝脏用作正常对照。灌注但未保存的肝脏用作再灌注对照。通过苏木精-伊红染色评估实质损伤。分别使用苦味酸天狼星红和戈登-斯威茨染色观察胶原和网状纤维网络。使用白蛋白免疫组织化学测定作为功能测试。冷保存步骤后，肝细胞肿胀，细胞核周围有“轻度空晕”，肝索保存完好，有许多圆形内皮细胞。向UW溶液中添加NPNa或GSNO可避免这些改变。保存48小时后再灌注的肝脏在中央静脉周围显示出扩大的空泡化区域，许多内皮细胞呈圆形并位于窦状隙腔内。胶原网络紊乱，而网状纤维网络变化较小。白蛋白优先分布在中央周围区域。相反，在NPNa或GSNO存在下保存的肝脏未显示空泡化，胶原和网状纤维网络均未改变。两组中白蛋白分布更均匀。总之，添加500微摩尔NPNa或100微摩尔GSNO作为NO供体，可改善UW溶液的特性，通过维持肝脏形态并避免冷保存/再灌注后的肝损伤来保存大鼠肝脏。
