Pavlock G S, Southard J H, Starling J R, Belzer F O
Cryobiology. 1984 Oct;21(5):521-8. doi: 10.1016/0011-2240(84)90050-6.
This study investigated lysosomal disruption during hypothermic perfusion preservation of kidneys and its possible relationship to viability. The percentage of free and bound enzyme activity was analyzed for three lysosomal enzymes in homogenates made from perfused canine kidney cortex tissue, including beta-glucuronidase, cathepsin-D, and aryl sulfatase. All three enzymes displayed characteristic increases in free enzyme activity (47-68%) throughout 5 days of perfusion preservation. The increased activity obtained at 5 days of preservation was found to indicate "severe" tissue damage, as shown by a similar increase obtained in renal cortex tissue exposed to warm ischemia (37 degrees C) for 4 hr or longer. Aryl sulfatase was found to be the most sensitive indicator of severe damage. Pretreatment of kidney donors with methylprednisolone, a lysosomal stabilizer, was also studied in kidneys exposed to 5 days of perfusion. Pretreatment was found to reduce the percentage of free lysosomal enzyme activity following 5 days (nonviable) of perfusion to those levels normally obtained following 3-day (viable) perfusion. This indicates that methylprednisolone may be useful in modulating the severe disruption of lysosomes induced by long-term preservation. It is concluded that extensive disruption of lysosomes occurs during hypothermic perfusion preservation and may represent one cause for loss of organ viability.
本研究调查了肾脏低温灌注保存过程中的溶酶体破坏及其与生存能力的可能关系。分析了从灌注的犬肾皮质组织匀浆中提取的三种溶酶体酶（包括β-葡萄糖醛酸酶、组织蛋白酶-D和芳基硫酸酯酶）的游离和结合酶活性百分比。在整个5天的灌注保存过程中，所有三种酶的游离酶活性均呈现出特征性增加（47%-68%）。发现在保存5天时获得的活性增加表明存在“严重”组织损伤，这与暴露于温暖缺血（37℃）4小时或更长时间的肾皮质组织中观察到的类似增加情况一致。芳基硫酸酯酶被发现是严重损伤的最敏感指标。还对肾脏供体用溶酶体稳定剂甲泼尼龙进行预处理后的肾脏进行了5天灌注研究。结果发现，预处理可使灌注5天（无活力）后的游离溶酶体酶活性百分比降低至通常在灌注3天（有活力）后获得的水平。这表明甲泼尼龙可能有助于调节长期保存诱导的溶酶体严重破坏。研究得出结论，低温灌注保存过程中会发生广泛的溶酶体破坏，这可能是器官生存能力丧失的一个原因。
