Eubanks J W, Sabek O, Kotb M, Gaber L W, Henry J, Hijiya N, Britt L G, Gaber A O, Goyert S M
Department of Surgery, University of Tennessee, Memphis 38163, USA.
Ann Surg. 1998 Jun;227(6):904-11. doi: 10.1097/00000658-199806000-00014.
The purpose of this study was to determine whether pathologic progression and cytokine responses in acute pancreatitis (AP) are altered in the absence of endotoxemia.
Previous studies have demonstrated that AP is characterized by rapid production and release of inflammatory cytokines, which play a major role in the local pancreatic and systemic complications of this disease. Infection and endotoxemia have been implicated as a major source of morbidity and death in AP and as possible stimuli for the overwhelming cytokine response seen in this disease.
AP was induced by a choline-deficient and ethionine-supplemented diet for 4 days in normal C57BL/6J mice (controls, n = 23) and in CD14 knockout mice (CD14KO, n = 23), which cannot produce circulating cytokines in response to endotoxin. Control and endotoxin-resistant mice were killed at time 0, then at 24, 48, 72, and 96 hours after the start of the diet. At each time point serum was collected for amylase, glucose, and cytokine measurements (tumor necrosis factor-alpha [TNFalpha] and interleukin-1beta [IL1beta]), and the pancreas was removed for histologic examination. TNFalpha was measured with a bioassay using WEHI-2F cells and IL1beta with a bioassay using D10.G4.1 cells.
CD14KO mice developed biochemical manifestations of AP with alterations in amylase levels, hypoglycemia, weight loss, and histologic changes of pancreatitis similar to the pattern seen in control mice. TNFalpha and IL1beta production had similar kinetics in both groups, with significant peak TNFalpha serum levels at 72 hours and a progressive rise of IL1beta levels throughout the study period. Histologic changes appeared earlier and were more pronounced in the control versus the CD14KO mice. However, the mortality rate was identical (20% at 96 hours) for both groups.
These results demonstrate that the progression of AP, the cytokine response associated with the disease, and early death are independent of endotoxin action. These findings, which suggest that an uncharacterized stimulus is responsible for triggering the cytokine cascade in this disease, may have significant implications for the management of patients with AP.
本研究旨在确定在无内毒素血症的情况下，急性胰腺炎（AP）的病理进展和细胞因子反应是否会发生改变。
先前的研究表明，AP的特征是炎症细胞因子迅速产生和释放，这些细胞因子在该疾病的局部胰腺和全身并发症中起主要作用。感染和内毒素血症被认为是AP发病和死亡的主要原因，也是该疾病中所见的压倒性细胞因子反应的可能刺激因素。
在正常C57BL/6J小鼠（对照组，n = 23）和CD14基因敲除小鼠（CD14KO，n = 23）中，通过给予胆碱缺乏和补充乙硫氨酸的饮食4天来诱导AP，CD14KO小鼠不能产生对内毒素作出反应的循环细胞因子。在时间0、然后在饮食开始后的24、48、72和96小时处死对照小鼠和抗内毒素小鼠。在每个时间点收集血清以测量淀粉酶、葡萄糖和细胞因子（肿瘤坏死因子-α [TNFα]和白细胞介素-1β [IL1β]），并取出胰腺进行组织学检查。使用WEHI-2F细胞通过生物测定法测量TNFα，使用D10.G4.1细胞通过生物测定法测量IL1β。
CD14KO小鼠出现了AP的生化表现，淀粉酶水平改变、低血糖、体重减轻以及胰腺炎的组织学变化，与对照小鼠所见模式相似。两组中TNFα和IL1β的产生具有相似的动力学，TNFα血清水平在72小时达到显著峰值，并在整个研究期间IL1β水平逐渐升高。组织学变化在对照小鼠中比在CD14KO小鼠中出现得更早且更明显。然而，两组的死亡率相同（96小时时为20%）。
这些结果表明，AP的进展、与该疾病相关的细胞因子反应以及早期死亡均与内毒素作用无关。这些发现表明，一种未明确的刺激物是引发该疾病中细胞因子级联反应的原因，这可能对AP患者的管理具有重要意义。
