Alam H B, Austin B, Koustova E, Rhee P
Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.
J Am Coll Surg. 2001 Sep;193(3):255-63. doi: 10.1016/s1072-7515(01)01004-3.
Resuscitation with Lactated Ringer's solution after hemorrhagic shock in rats has been shown to cause early cellular injury in the lung. We hypothesized that the use of energy substrates, such as ketone bodies, in the resuscitation fluids would protect against this injury. As markers of cellular injury we measured the induction of apoptotic cell death and the expression of Intracellular Adhesion Molecule-1 (ICAM-1).
Male Sprague Dawley rats (n = 35) under inhaled isoflurane anesthesia had placement of femoral arterial and venous catheters. A three-stage hemorrhage model was used for this experiment. There was an initial hemorrhage of 27 mL/kg for 10 minutes. During the next 75 minutes another 8 mL/kg of blood was withdrawn at a steady rate. The resuscitation fluids were then infused for 45 minutes during which the third continuous hemorrhage of 8 mL/kg was performed. The animals were randomized to five groups: 1) sham hemorrhage (n = 6); 2) sham resuscitation (n = 7); 3) Lactated Ringer's resuscitation, three times the volume of shed blood (n = 8); 4) Ketone Ringer's (containing 28 mEq/L of beta-hydroxybutyrate) resuscitation, three times the volume of shed blood (n = 7); and 5) plasma resuscitation, volume equal to shed blood (n = 7). The animals were sacrificed 1 hour after resuscitation and lungs were harvested. Western blot technique was used for the determination of proapoptotic protein (bax), antiapoptotic protein (bcl-2), apoptotic fragments of poly ADP-ribose polymerase, and ICAM-1. Sections of lung were also subjected to immunostaining using antibodies to bax and ICAM-1 proteins (reported as number of positive cells/mm2).
Lactated Ringer's resuscitation caused a significant increase in pulmonary apoptosis and ICAM-1 expression compared with the sham hemorrhage group. Animals resuscitated with Ketone Ringer's solution and plasma did not show this injury pattern.
Substitution of lactate with ketone bodies in the resuscitation fluid attenuates the expression of cellular injury markers in the lung.
已表明大鼠失血性休克后用乳酸林格氏液复苏会导致肺部早期细胞损伤。我们推测在复苏液中使用能量底物（如酮体）可预防这种损伤。作为细胞损伤的标志物，我们检测了凋亡细胞死亡的诱导情况以及细胞间黏附分子 -1（ICAM -1）的表达。
吸入异氟醚麻醉下的雄性斯普拉格 - 道利大鼠（n = 35）植入股动脉和静脉导管。本实验采用三阶段出血模型。初始阶段以27 mL/kg的量出血10分钟。在接下来的75分钟内，以稳定速率再抽取8 mL/kg的血液。然后输注复苏液45分钟，在此期间进行第三次持续出血，出血量为8 mL/kg。动物被随机分为五组：1）假出血组（n = 6）；2）假复苏组（n = 7）；3）乳酸林格氏液复苏组，输注量为失血量的三倍（n = 8）；4）酮体林格氏液（含28 mEq/L的β - 羟基丁酸）复苏组，输注量为失血量的三倍（n = 7）；5）血浆复苏组，输注量与失血量相等（n = 7）。复苏后1小时处死动物并摘取肺脏。采用蛋白质免疫印迹技术测定促凋亡蛋白（bax）、抗凋亡蛋白（bcl -2）、聚ADP - 核糖聚合酶凋亡片段以及ICAM -1。肺组织切片也用针对bax和ICAM -1蛋白的抗体进行免疫染色（以每平方毫米阳性细胞数报告）。
与假出血组相比，乳酸林格氏液复苏导致肺部凋亡和ICAM -1表达显著增加。用酮体林格氏液和血浆复苏的动物未表现出这种损伤模式。
复苏液中用酮体替代乳酸可减轻肺部细胞损伤标志物的表达。
