Guan Jian, Jin Da-di, Jin Li-juan, Lu Qi
Department of Orthopaedics, NanFang Hospital of the First Military Medical University, Guangzhou, People's Republic of China.
J Trauma. 2002 Jan;52(1):104-11. doi: 10.1097/00005373-200201000-00018.
Apoptosis in organs of rats in early stage after polytrauma combined with shock was researched.
Sixty Sprague-Dawley rats were divided into six groups: normal control (A, n = 6), sham-operation (N, n = 6), single hemorrhagic shock (S, n = 6), two-site trauma/shock (B, n = 6), four-site trauma/shock (C, n = 6), and six-site trauma/shock (D, n = 30). Shock was kept 60 min by blood withdrawal. Polytrauma was performed by clamping different sites of limbs to make fractures according to different groups: B at both femurs; C at femurs and tibias; and D at femurs, tibias, and humeri. The animals of A were totally normal without any operation. The rats of N, S, B, and C were killed at 6 hours after resuscitation, and the rats of D were killed at 1, 3, 6, 12, and 24 hours, respectively. Then, DNA agarose gel electrophoresis, in situ end-labeling (ISEL), and light and electron microscopy were performed and the percentage of DNA fragmentation was detected to assess apoptosis.
In B, C, and D, the special ladder patterns for apoptosis were seen in thymus, spleen, liver, lung, and intestine, but not in heart, kidney, and brain. However, positive responses were observed in all these eight organs by ISEL. At 6 hours after resuscitation, the percentages of DNA fragmentation in thymus, spleen, liver, lung, and intestine all increased together with the severity of trauma. In D, the percentages of DNA fragmentation in these five organs all increased significantly at 1 hour after resuscitation. At 3 hours, the percentages in spleen, liver, lung, and intestine reached peak, and declined gradually afterward, whereas those in thymus continued increasing after 3 hours and kept stable from 6 hours to 24 hours. It was shown by morphologic examination that the majority of apoptotic cells lay in cortex of thymus, in growth center of white pulp of spleen, in border area of hepatic lobule and portal area of liver, and at the base of crypts of intestine. In lung, multiple kinds of cells, including alveolar epithelial cells, vascular endothelial cells, and polymorphonuclear neutrophils, induced apoptosis.
Apoptosis was induced in thymus, spleen, liver, lung, and intestine in early stage after polytrauma combined with shock, which may play partial roles in the development of multiple organ failure.
研究多发伤合并休克大鼠早期器官的细胞凋亡情况。
将60只Sprague-Dawley大鼠分为6组:正常对照组(A组,n = 6)、假手术组(N组,n = 6)、单纯失血性休克组(S组,n = 6)、两处创伤/休克组(B组,n = 6)、四处创伤/休克组(C组,n = 6)和六处创伤/休克组(D组,n = 30)。通过放血使休克持续60分钟。根据不同组别的情况,通过钳夹四肢不同部位造成骨折来实施多发伤:B组为双侧股骨;C组为股骨和胫骨;D组为股骨、胫骨和肱骨。A组动物完全正常,未进行任何手术。N组、S组、B组和C组的大鼠在复苏后6小时处死,D组的大鼠分别在复苏后1、3、6、12和24小时处死。然后进行DNA琼脂糖凝胶电泳、原位末端标记(ISEL)以及光镜和电镜检查,并检测DNA片段化百分比以评估细胞凋亡情况。
在B组、C组和D组中,胸腺、脾脏、肝脏、肺和肠道出现了细胞凋亡的特征性梯状条带,但心脏、肾脏和大脑未出现。然而,ISEL在所有这八个器官中均观察到阳性反应。复苏后6小时,胸腺、脾脏、肝脏、肺和肠道的DNA片段化百分比均随着创伤严重程度的增加而升高。在D组中,复苏后1小时这五个器官的DNA片段化百分比均显著升高。3小时时,脾脏、肝脏、肺和肠道的百分比达到峰值,随后逐渐下降,而胸腺的百分比在3小时后继续升高,并在6小时至24小时保持稳定。形态学检查显示,大多数凋亡细胞位于胸腺皮质、脾脏白髓生发中心、肝小叶边缘区和肝脏门管区以及肠隐窝底部。在肺中,多种细胞,包括肺泡上皮细胞、血管内皮细胞和多形核中性粒细胞,诱导了细胞凋亡。
多发伤合并休克大鼠早期胸腺、脾脏、肝脏、肺和肠道诱导了细胞凋亡,这可能在多器官功能障碍的发生中起部分作用。