Song Tianzhang, Yin Hongling, Chen Jintao, Huang Lilin, Jiang Juan, He Tailong, Huang Huaiqiu, Hu Xuchu
Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, China; Education Ministry Key Laboratory for Tropical Disease Control Research, Sun Yat-sen University, Guangzhou, Guangdong, China.
Department of Clinical Laboratory Medicine, The Third Affiliated Hospital of Guangzhou Medical University, Guangdong, China.
J Surg Res. 2016 Jun 15;203(2):476-82. doi: 10.1016/j.jss.2016.03.019. Epub 2016 Mar 17.
Cecal ligation and puncture (CLP) is the most commonly used model to simulate human polymicrobial sepsis. However, the severity of CLP is difficult to be standardized across different laboratories. The aim of the present study was to evaluate the influence of ligated cecal volume and length on mortality in mouse CLP model.
Cecal length and volume were measured from 120 Kunming mice subjected to CLP or sham operation. According to cecal volume, mice were divided into three groups, volume0.0∼0.2 (0.0 cm(3)-0.2 cm(3)), volume0.2∼0.4 (0.2 cm(3)-0.4 cm(3)), and volume>0.4 (larger than 0.4 cm(3)). The contents of cytokines, including interleukin-1β, interleukin-6, and TNF-α, were measured at 3 h after surgery. The blood bacterial load and oxidative stress indicators (including malondialdehyde and superoxide dismutase) were measured at 12 h after surgery.
There was no significant difference on 72-h survival rate between the mice with cecum longer than 2 cm and shorter than 2 cm. Compared to the other volume groups, volume>0.4 group showed significantly increased blood bacterial load, malondialdehyde levels in lung and liver, and pro-inflammatory cytokines in serum. Surprisingly, the survival rate in volume>0.4 (0%) group showed significant difference from those of volume0.0∼0.2 group (40%) and volume0.2∼0.4 group (40%).
The mice in volume>0.4 group have much serious inflammatory reaction and are easier to die. As the proportion of volume>0.4 mice is near 20%, it can have large influence on most of the related studies using this CLP model.
盲肠结扎穿刺术(CLP)是模拟人类多微生物败血症最常用的模型。然而,不同实验室之间CLP的严重程度难以标准化。本研究的目的是评估结扎盲肠的体积和长度对小鼠CLP模型死亡率的影响。
对120只接受CLP或假手术的昆明小鼠测量盲肠长度和体积。根据盲肠体积,将小鼠分为三组,体积0.0∼0.2(0.0立方厘米 - 0.2立方厘米)、体积0.2∼0.4(0.2立方厘米 - 0.4立方厘米)和体积>0.4(大于0.4立方厘米)。术后3小时测量细胞因子(包括白细胞介素-1β、白细胞介素-6和肿瘤坏死因子-α)的含量。术后12小时测量血细菌载量和氧化应激指标(包括丙二醛和超氧化物歧化酶)。
盲肠长度大于2厘米和小于2厘米的小鼠之间72小时存活率无显著差异。与其他体积组相比,体积>0.4组血细菌载量、肺和肝脏中的丙二醛水平以及血清中的促炎细胞因子显著增加。令人惊讶的是,体积>0.4(0%)组的存活率与体积0.0∼0.2组(40%)和体积0.2∼0.4组(40%)有显著差异。
体积>0.4组的小鼠炎症反应更严重,更容易死亡。由于体积>0.4的小鼠比例接近20%,这可能对大多数使用该CLP模型的相关研究产生很大影响。