Montero Edna F S, Abrahão Marcos S, Koike Márcia K, Manna Mônica C B, Ramalho Carlos E B
Disciplina de Técnica Operatória e Cirurgia Experimental, Departamento de Cirurgia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
Microsurgery. 2003;23(5):517-21. doi: 10.1002/micr.10163.
Our objective was to evaluate intestinal ischemia-reperfusion injury in growing rats, modulated by hypothermia (I/RH) and N-acetylcysteine (NAC). We used 30 EPM-1 Wistar male rats, aged around 35 days, weighing 90 g. Rats were randomized into 5 groups with 6 animals in each: I/RH group, intestinal ischemia under hypothermia for 40 min and reperfusion for 30 min; I/RH-NAC group, same procedure but adding NAC (150 mg x kg(-1)), previously with ischemia; S-H group, topic hypothermia for 40 min, and observation for 30 min; I/R H-Ve group; and S-NAC group, NAC administration and observation for 70 min. All animals were heparinized and anesthetized with ketamine (60 mg kg(-1)) and xylazine (10 mg kg(-1)) intramuscularly. Surgical procedures were done under microsurgical technique (augmentation, 10x). After laparotomy, the superior mesenteric artery was dissected and clamped to promote ischemia. Topic hypothermia was obtained by using plastic bags at 4 degrees C, changed every 10 min. Rats were sacrificed by exsanguination, and blood samples were utilized to measure D(-)lactate. Intestinal fragments were removed for morphological study. Statistical analysis was done with nonparametric tests (P <or= 0.05). Concerning to D(-)lactate, the data showed biochemical tissue injury, with hypothermia only (S-H = 27 mg/dl), and this became more important when intestinal ischemia and reperfusion were associated to hypothermia (I/RH = 36 mg/dl). NAC decreased ischemia-reperfusion injury (I/RH-NAC = 19 mg/dl). Morphologic tissue injuries, evaluated by hematoxylin-eosin staining, showed grades 4 and 5 for the I/RH and I/RH-Ve groups, respectively, in contrast with other groups (I/RH-NAC = 2, S-H = 1, and S-NAC = 1). Based on our data, we conclude that intestinal ischemia reperfusion injury occurred morphologically as well as functionally, even under hypothermia. However, NAC showed a protective effect on the small bowel from ischemia-reperfusion injury.
我们的目的是评估低温(I/RH)和N-乙酰半胱氨酸(NAC)对生长中大鼠肠道缺血再灌注损伤的影响。我们使用了30只EPM-1品系的Wistar雄性大鼠,年龄约35天,体重90克。大鼠被随机分为5组,每组6只:I/RH组,低温下肠道缺血40分钟,再灌注30分钟;I/RH-NAC组,相同程序,但在缺血前添加NAC(150毫克/千克);S-H组,局部低温40分钟,观察30分钟;I/R H-Ve组;以及S-NAC组,给予NAC并观察70分钟。所有动物均经肝素化处理,并通过肌肉注射氯胺酮(60毫克/千克)和赛拉嗪(10毫克/千克)进行麻醉。手术操作在显微外科技术(放大倍数为10倍)下进行。剖腹术后,解剖并夹闭肠系膜上动脉以造成缺血。通过使用4℃的塑料袋实现局部低温,每10分钟更换一次。大鼠通过放血处死,采集血样以测量D(-)乳酸。取出肠道片段进行形态学研究。采用非参数检验进行统计分析(P≤0.05)。关于D(-)乳酸,数据显示仅低温时(S-H = 27毫克/分升)存在生化组织损伤,而当肠道缺血和再灌注与低温相关时(I/RH = 36毫克/分升),这种损伤变得更为严重。NAC减轻了缺血再灌注损伤(I/RH-NAC = 19毫克/分升)。通过苏木精-伊红染色评估的形态学组织损伤显示,I/RH组和I/RH-Ve组分别为4级和5级,与其他组(I/RH-NAC = 2级,S-H = 1级,S-NAC = 1级)不同。基于我们的数据,我们得出结论,即使在低温情况下,肠道缺血再灌注损伤在形态和功能上均会发生。然而,NAC对小肠缺血再灌注损伤具有保护作用。