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中华危重症医学杂志(电子版)

中华危重症医学杂志(电子版) ›› 2021, Vol. 14 ›› Issue (01) : 15 -19. doi: 10.3877/cma.j.issn.1674-6880.2021.01.003

所属专题： 文献；

免疫衰老与脓毒症

免气腹单孔腹腔镜辅助盲肠结扎穿孔术建立脓毒症大鼠模型
杨茂宪1, 王倩倩1, 沈鹏1, 徐龙生2, 张小平3, 蔚文龙1, 钟林1, 戴靓潇1, 朱建刚1, 施云超1,†()   
  1. 1. 314001　浙江嘉兴，嘉兴市第一医院（嘉兴学院附属医院）重症医学科
    2. 314001　浙江嘉兴，嘉兴市第一医院（嘉兴学院附属医院）中心实验室
    3. 314001　浙江嘉兴，嘉兴市第一医院（嘉兴学院附属医院）科教科
  • 收稿日期:2020-12-14 出版日期:2021-02-28
  • 通信作者: 施云超
  • 基金资助:
    嘉兴市医学重点学科重症医学（支撑学科）(2019-ZC-12); 浙江省嘉兴市科技计划项目(2018AY32017); 嘉兴市医学领军人才培养人选(2019-lj-03)

Establishment of a rat model of sepsis by cecal ligation and puncture using single-port gasless laparoscopy

Maoxian Yang1, Qianqian Wang1, Peng Shen1, Longsheng Xu2, Xiaoping Zhang3, Wenlong Wei1, Lin Zhong1, Liangxiao Dai1, Jiangang Zhu1, Yunchao Shi1,†()   

  1. 1. Department of Intensive Care Unit, the First Hospital of Jiaxing (Affiliated Hospital of Jiaxing University), Jiaxing 314001, China
    2. Central Laboratory, the First Hospital of Jiaxing (Affiliated Hospital of Jiaxing University), Jiaxing 314001, China
    3. Department of Science and Education, the First Hospital of Jiaxing (Affiliated Hospital of Jiaxing University), Jiaxing 314001, China
  • Received:2020-12-14 Published:2021-02-28
  • Corresponding author: Yunchao Shi
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杨茂宪, 王倩倩, 沈鹏, 徐龙生, 张小平, 蔚文龙, 钟林, 戴靓潇, 朱建刚, 施云超. 免气腹单孔腹腔镜辅助盲肠结扎穿孔术建立脓毒症大鼠模型[J]. 中华危重症医学杂志(电子版), 2021, 14(01): 15-19.

Maoxian Yang, Qianqian Wang, Peng Shen, Longsheng Xu, Xiaoping Zhang, Wenlong Wei, Lin Zhong, Liangxiao Dai, Jiangang Zhu, Yunchao Shi. Establishment of a rat model of sepsis by cecal ligation and puncture using single-port gasless laparoscopy[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2021, 14(01): 15-19.

目的

基于免气腹单孔腹腔镜辅助盲肠结扎穿孔（CLP）术建立脓毒症大鼠模型。

方法

将64只大鼠分为A组（传统CLP术对照组）、B组（免气腹单孔腹腔镜辅助下CLP术对照组）、C组（传统CLP术穿孔组）及D组（免气腹单孔腹腔镜辅助下CLP术穿孔组），每组各16只。记录建模后24 h各组大鼠的死亡情况，并检测各组大鼠血清白细胞介素1β（IL-1β）、IL-6、IL-10及肿瘤坏死因子α（TNF-α）水平。同时，采用苏木素-伊红（HE）染色法观察各组大鼠肺组织、小肠组织、肾组织的病理变化。

结果

建模后24 h，A组、B组大鼠均存活，C组大鼠死亡7只，D组大鼠死亡8只。各组大鼠间血清IL-1β、IL-6、IL-10、TNF-α水平比较，差异均有统计学意义（F=21.461、27.372、51.113、33.799，P均<0.001）。进一步两两比较发现，C组、D组大鼠的血清IL-1β[（7.4 ± 2.7）、（7.4 ± 2.2）、（2.4 ± 0.6）、（2.4 ± 0.6）ng/L]，IL-6[（2.05 ± 0.46）、（2.14 ± 0.58）、（0.82 ± 0.20）、（0.86 ± 0.16）ng/L]，IL-10[（59 ± 10）、（61 ± 13）、（19 ± 6）、（22 ± 6）ng/L]，TNF-α[（64 ± 13）、（74 ± 21）、（24 ± 4）、（23 ± 5）ng/L]水平较A组、B组大鼠均显著升高（P均<0.05），而A组、B组大鼠间及C组、D组大鼠间血清IL-1β、IL-6、IL-10、TNF-α水平比较，差异均无统计学意义（P均> 0.05）。HE染色发现，A组、B组大鼠肺组织、小肠组织、肾组织结构完整，未见炎症细胞浸润；C组、D组可见肺泡结构完整性破环、肺泡壁增厚、炎症细胞浸润，部分肠绒毛上皮细胞坏死脱落，部分肾小管上皮细胞水肿。

结论

利用免气腹单孔腹腔镜能简单、有效地建立大鼠CLP术脓毒症模型。

关键词: 脓毒症, 盲肠结扎穿孔, 腹腔镜, 大鼠
Objective

To establish a rat model of sepsis by cecal ligation and puncture (CLP) using single-port gasless laparoscopy.

Methods

Sixty-four rats were divided into the group A (conventional CLP control group), group B (single-port gasless laparoscopy assisted CLP control group), group C (conventional CLP group) and group D (single-port gasless laparoscopy assisted CLP group), 16 rats in each group. After 24 h of model replication, the death of rats was recorded and the levels of interleukin-1beta (IL-1β), IL-6, IL-10 and tumor necrosis factor-alpha (TNF-α) were measured and compared in each group. The pathological changes of lung, small intestine and kidney of rats were observed by hematoxylin-eosin (HE) staining.

Results

All rats in groups A and B survived, while seven rats in the group C and eight rats in the group D died at 24 h after modeling. The levels of IL-1β, IL-6, IL-10 and TNF-α among the four groups all showed significant differences (F=21.461, 27.372, 51.113, 33.799; all P<0.001). Further pairwise comparison revealed that the levels of IL-1β [(7.4 ± 2.7), (7.4 ± 2.2), (2.4 ± 0.6), (2.4 ± 0.6) ng/L], IL-6 [(2.05 ± 0.46), (2.14 ± 0.58), (0.82 ± 0.20), (0.86 ± 0.16) ng/L], IL-10 [(59 ± 10), (61 ± 13), (19 ± 6), (22 ± 6) ng/L] and TNF-α [(64 ± 13), (74 ± 21), (24 ± 4), (23 ± 5) ng/L] in groups C and D were significantly higher than those in groups A and B (all P < 0.05). However, there was no significant difference in IL-1β, IL-6, IL-10 and TNF-α levels between rats in groups A and B and between rats in groups C and D (all P > 0.05). HE staining revealed that the lung tissue, small intestine tissue and kidney tissue of rats in groups A and B were structurally intact and no inflammatory cell infiltration was observed. Meanwhile, in groups C and D, the structural integrity of alveoli was damaged, alveolar walls were thickened, and inflammatory cell infiltrated; some intestinal villus epithelial cells were necrotic and exfoliated; some renal tubular epithelial cells were edematous.

Conclusion

The use of single-port gasless laparoscopy combined with CLP can establish a simple and effective model of sepsis in rats.

Key words: Sepsis, Cecal ligation and puncture, Laparoscopes, Rats
表1 4组大鼠血清IL-1β、IL-6、IL-10、TNF-α浓度的比较（ng/L，±s）
组别 只数 IL-1β IL-6 IL-10 TNF-α
A组 8 2.4 ± 0.6 0.82 ± 0.20 19 ± 6 24 ± 4
B组 8 2.4 ± 0.6 0.86 ± 0.16 22 ± 6 23 ± 5
C组 8 7.4 ± 2.7ab 2.05 ± 0.46ab 59 ± 10ab 64 ± 13ab
D组 8 7.4 ± 2.2ab 2.14 ± 0.58ab 61 ± 13ab 74 ± 21ab
F值   21.461 27.372 51.113 33.799
P值   < 0.001 < 0.001 < 0.001 < 0.001
图2 4组大鼠肺脏、小肠、肾脏的病理组织学变化
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