Jung S I, Chang G J, Corbascio M, Potts M, Bedolli M, Ascher N I, Freise C F
Department of Surgery, Korea University, Seoul, South Korea.
J Surg Res. 2001 Sep;100(1):69-75. doi: 10.1006/jsre.2001.6219.
Prolonged cold ischemia has been shown to be an important factor in the development of posttransplant renal dysfunction. The exact mechanisms have not been completely defined. The expression of intercellular adhesion molecule-1 (ICAM-1) (CD 54) in rat kidneys stored in University of Wisconsin (UW) solution was studied in an attempt to correlate ischemia time with immunogenicity of the graft.
Kidneys from male Lewis rats were perfused with UW solution, removed, and bathed in UW solution at 4 degrees C for 4, 12, 24, and 48 h. For the evaluation of expression of ICAM-1, immunohistochemical staining, Western blotting, and semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) were performed.
Immunohistochemical staining in normal, nonischemic kidneys revealed that glomerular capillaries expressed ICAM-1 but that tubular cells did not. The preserved kidneys were analyzed by immunohistochemistry, Western blotting, and semiquantitative RT-PCR and showed increased transcription and expression of ICAM-1 in the cortex of the kidney. Expression reached a maximum at 24 h and declined at 48 h. The ICAM-1 protein expression in the preserved kidney cortex relative to control kidneys was increased at 4 h (1.68 +/- 0.60-fold of control kidneys, P = 0.06), 12 h (2.38 +/- 0.90-fold, P = 0.02), 24 h (3.70 +/- 1.29-fold, P = 0.01), and 48 h (2.00 +/- 0.54-fold, P = 0.01). The messenger RNA expression (the ratio of ICAM-1 to glyceraldehyde-3-phosphate dehydrogenase) in preserved kidneys cortex relative to control kidneys was increased at 4 h (1.19 +/- 0.14-fold of control kidneys), 12 h (1.38 +/- 0.16-fold), 24 h (1.77 +/- 0.29-fold), and 48 h (1.19 +/- 0.12-fold) (P < 0.05 for all time points).
We conclude that cold preservation of rat kidneys in UW solution induces increasing levels of ICAM-1 cell surface expression and gene transcription. Further study is necessary to determine if this increase in adhesion molecule expression increases the immunogenicity of the allograft and contributes to the development of posttransplant renal dysfunction.
长期冷缺血已被证明是移植后肾功能障碍发生发展的一个重要因素。确切机制尚未完全明确。本研究旨在探讨威斯康星大学（UW）溶液保存的大鼠肾脏中细胞间黏附分子-1（ICAM-1，CD 54）的表达情况，试图将缺血时间与移植物的免疫原性相关联。
用UW溶液灌注雄性Lewis大鼠的肾脏，取出后于4℃在UW溶液中浸泡4、12、24和48小时。采用免疫组织化学染色、蛋白质印迹法及半定量逆转录聚合酶链反应（RT-PCR）评估ICAM-1的表达。
正常非缺血肾脏的免疫组织化学染色显示，肾小球毛细血管表达ICAM-1，而肾小管细胞不表达。对保存的肾脏进行免疫组织化学、蛋白质印迹法及半定量RT-PCR分析，结果显示肾脏皮质中ICAM-1的转录和表达增加。表达在24小时达到峰值，48小时下降。保存的肾脏皮质中ICAM-1蛋白表达相对于对照肾脏在4小时（为对照肾脏的1.68±0.60倍，P = 0.06）、12小时（2.38±0.90倍，P = 0.02）、24小时（3.70±1.29倍，P = 0.01）和48小时（2.00±0.54倍，P = 0.01）时升高。保存的肾脏皮质中信使核糖核酸表达（ICAM-1与甘油醛-3-磷酸脱氢酶的比值）相对于对照肾脏在4小时（为对照肾脏的1.19±0.14倍）、12小时（1.38±0.16倍）、24小时（1.77±0.29倍）和48小时（1.19±0.12倍）时升高（所有时间点P<0.05）。
我们得出结论，UW溶液中大鼠肾脏的冷保存诱导ICAM-1细胞表面表达和基因转录水平升高。有必要进一步研究黏附分子表达的这种增加是否会增加同种异体移植物的免疫原性并导致移植后肾功能障碍的发生。
