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

doi:10.3877/cma.j.issn.1674-6880.2021.01.003

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. 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. 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/OL]. 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/OL]. 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 ± 2.7^ab	2.05 ± 0.46^ab	59 ± 10^ab	64 ± 13^ab
D组	8	7.4 ± 2.2^ab	2.14 ± 0.58^ab	61 ± 13^ab	74 ± 21^ab
F值		21.461	27.372	51.113	33.799
P值		< 0.001	< 0.001	< 0.001	< 0.001

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 ± 2.7^ab	2.05 ± 0.46^ab	59 ± 10^ab	64 ± 13^ab
D组	8	7.4 ± 2.2^ab	2.14 ± 0.58^ab	61 ± 13^ab	74 ± 21^ab
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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