Sepsis modeling in mice: ligation length is a major severity factor in cecal ligation and puncture.
Sepsis modeling in mice: ligation length is a major severity factor in cecal ligation and puncture.
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DOI：
10.1186/s40635-016-0096-z
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发表时间：
2016-12
影响因子：
3.5
通讯作者：
Minville V
中科院分区：
文献类型：
--
作者：
Ruiz S;Vardon-Bounes F;Merlet-Dupuy V;Conil JM;Buléon M;Fourcade O;Tack I;Minville V
盲肠结扎穿孔（CLP）模型是脓毒症研究的金标准，与死亡率的重要变异性相关。虽然在CLP动物研究中很好地描述了穿刺次数和针头尺寸，但盲肠结扎的长度通常没有。盲肠结扎与小鼠存活率之间的关系在文献中有简要报道;因此，我们设计了一项小鼠研究，研究三种标准化盲肠结扎长度对死亡率和随后败血症严重程度的影响。雄性C57 BL/6 J小鼠接受标准化CLP。盲肠在其总长度的5%、20%或100%处结扎，并通过单次20-G穿刺进一步穿孔。分析死亡率。我们评估了对照组和结扎小鼠术后的血乳酸、血清肌酐水平和血清细胞因子（TNF-α、IL-1β、IL-6和IL-10）。死亡率与结扎长度直接相关：“100%”组的中位存活时间为24小时，“20%”组为44小时。血乳酸随结扎时间延长而增加。术后6小时，结扎组的促炎细胞因子显着增加，其中100%组的血清IL-6水平显着高于其他结扎组。20%组表现出脓毒性休克的特征，低血压低于65 mmHg，促炎平衡，器官功能障碍和高乳酸血症。盲肠结扎长度似乎是小鼠CLP模型中的主要限制因素。因此，这个实验模型应该在未来的协议设计中具有高度的一致性。
The cecal ligation and puncture (CLP) model, a gold standard in sepsis research, is associated with an important variability in mortality. While the number of punctures and needle size is well described in CLP animal studies, the length of cecal ligation is often not. The relationship between cecal ligation and survival in mice is briefly reported in the literature; therefore, we devised an investigation in mice of the consequences of three standardized cecal ligation lengths on mortality and the severity of the ensued sepsis. Male C57BL/6J mice underwent standardized CLP. The cecum was ligated at 5, 20, or 100 % of its total length and further perforated by a single 20-G puncture. Mortality was analyzed. We assessed blood lactate, serum creatinine levels, and serum cytokines (TNF-α, IL-1β, IL-6, and IL-10) after procedure in a control group and in ligated mice. Mortality was directly related to ligation length: median survival was 24 h for the “100 %” group and 44 h for the “20 %” group. Blood lactate increased proportionally with the ligation length. At 6 h post-procedure, pro-inflammatory cytokines significantly increased in the ligated group with significantly higher serum levels of IL-6 in the 100 % group compared to the other ligated groups. The 20 % group exhibited the characteristics of septic shock with hypotension below 65 mmHg, pro-inflammatory balance, organ dysfunction, and hyperlactatemia. Cecal ligation length appears to be a major limiting factor in the mouse CLP model. Thus, this experimental model should be performed with high consistency in future protocol designs.