Budziński G, Wiaderkiewicz R, Caban A, Suszka-Świtek A, Oczkowicz G, Wystrychowski W, Heitzman M, Dolińska B, Ryszka F, Cierpka L
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland.
Transplant Proc. 2011 Oct;43(8):2900-2. doi: 10.1016/j.transproceed.2011.08.066.
Organ ischemia is accompanied by cell death due to apoptosis. It occurs together with necrosis, which has more unfavorable consequences due to the release of cytokines that activate the inflammatory response cascade. The aim of this study was to assess the degree of apoptosis in porcine livers preserved by simple hypothermia for 12 hours using standard solutions (University of Wisconsin [UW] and histidine-tryptophan-glutarate [HTK]), and to evaluate the effect of prolactin (PRL) addition to the HTK solution.
The study was performed on the livers of Great White breed pigs, after inducing 30 minutes of warm ischemia (WIT30), followed by 30 minutes of perfusion-cooling to 4°C, and 12 hours of preservation. Livers were evaluated after preservation in Ringer's solution (control); UW (control reference fluid); HTK and HTK modified by the addition of prolactin (20 UI/L. Apoptosis was assessed in liver sections by the TdT-mediated dUTP nick-end labeling method after 12-hour preservation. We adopted a prevalence scale ranging from 0 to 3+, depending on the number of observed nuclei and apoptotic bodies (AB).
Preservation in Ringer's solution yielded AB distribution at the 1+ level, with a lack of characteristic localization resulting from necrotic lesions. Analysis of the livers preserved in the UW solution showed high, 3+ level of AB presence. For the tested HTK solution, the observed ABs localization value was 3+, whereas in the PRL-modified group it was also 3+, but with a tendency to move from zone II to cluster III, which is important for liver metabolic functions.
PRL improved the preservation properties of HTK for porcine livers by maintaining a high apoptosis level. It may stabilize cell membranes thus reducing the oncotic necrosis, promoting increased apoptosis during simple hypothermia.
器官缺血会伴随细胞因凋亡而死亡。它与坏死同时发生，坏死由于细胞因子的释放激活炎症反应级联，会产生更不利的后果。本研究的目的是评估使用标准溶液（威斯康星大学溶液[UW]和组氨酸-色氨酸-谷氨酸盐[HTK]）通过简单低温保存12小时的猪肝中的凋亡程度，并评估向HTK溶液中添加催乳素（PRL）的效果。
本研究在大白猪的肝脏上进行，先诱导30分钟的热缺血（WIT30），随后进行30分钟的灌注冷却至4°C，并保存12小时。肝脏在林格氏液（对照）、UW（对照参考液）、HTK以及添加了催乳素（20 UI/L）的改良HTK中保存后进行评估。在12小时保存后，通过TdT介导的dUTP缺口末端标记法评估肝切片中的凋亡情况。根据观察到的细胞核和凋亡小体（AB）数量，我们采用了从0到3+的发生率量表。
在林格氏液中保存导致AB分布处于1+水平，由于坏死性病变缺乏特征性定位。对在UW溶液中保存的肝脏分析显示AB存在水平较高，为3+。对于测试的HTK溶液，观察到的AB定位值为3+，而在PRL改良组中也是3+，但有从II区转移到III区的趋势，这对肝脏代谢功能很重要。
PRL通过维持较高的凋亡水平改善了HTK对猪肝的保存特性。它可能稳定细胞膜，从而减少肿胀性坏死，促进简单低温期间凋亡增加。
