中国实验动物学报2024，Vol.32Issue(4)：423-434,12.DOI:10.3969/j.issn.1005-4847.2024.04.002
基于NLRP3/Caspase-1/GSDMD焦亡通路探索小鼠急性肺损伤动态时间模型的建立
Establishment of a dynamic model of acute lung injury in mice based on the NLRP3/Caspase-1/gasdermin D signaling pyroptosis pathway
摘要
Abstract
Objective To establish a dynamic model of lipopolysaccharide-induced acute lung injury in mice based on the NLRP3/Caspase-1/gasdermin D(GSDMD)pyroptosis pathway,and observe the result ing lung injury at different time points.We aimed to identify the optimal time for modelling according to the injury at different time points and the expression of pyroptosis pathway-related proteins,to lay the foundation for animal models for subsequent experiments.Methods Fifty-four 6～8 weeks old male SPF BALB/c mice were divided randomly into nine groups,including Con group and model groups at 1,3,6,12,18,24,48,and 72 h.Body weight and lung tissue were detected by general and pathological observations and semi-quantitative scoring,including lung index,lung water content,and wet and dry weight ratio.The white blood cell count and concentrations of tumor necrosis factor-α,interleukin(IL)-6,IL-1β,IL-18,and BCA protein were detected in bronchoalveolar lavage fluid(BALF).The classic pyroptosis pathway-related proteins NLRP3,pro-Caspase 1,Caspase 1,and GSDMD were detected by Western Blot.Results Body weight decreased in all experimental groups,with the most significant weight loss in the 24 and 48 h groups.Gross observation and pathological examination of lung tissue showed that the most severe lung injury occurred at 24～72 h,with significant differences between each group and the control group.The lung index,lung water content,and wet/dry weight ratio were also significantly increased at 24～72 h.White blood cells in BALF started to increase from 6 h after model initiation,48 h can reach a peak,72 h all keep increasing.IL-18 in BALF began to increase at 24 h and continued to increase at 72 h.The inflammatory factors tumor necrosis factor-α,IL-1β,IL-6 were highest at 6 h and significantly reduced at 48 h.Protein concentrations in BALF were significantly increased within 24,48,and 72 h compared with those in the control group.The pyroptosis pathway proteins NLRP3,pro-Caspase-1,Caspase-1,and GSDMD were significantly enhanced in each time series,and channel protein expression was significantly enhanced at 24～72 h compared with that in the Con group.Conclusions Comprehensive analysis of experimental indicators,inflammatory factors,and pathway proteins at different times showed that the mechanism of pyroptosis was closely related to the occurrence and progression of acute lung injury.Expression of the pyroptosis pathway was most obvious and lung injury was most serious at 24～48 h.This study provides a model reference and experimental basis for subsequent studies of the specific mechanism and intervention targets of acute lung injury.关键词
焦亡/急性肺损伤/小鼠/动态时间模型/动物实验Key words
pyroptosis/acute lung injury/mice/dynamic time model/animal experiment分类
生物科学引用本文复制引用
樊懿萱,王心威,李军梅,易亮,杨志旭..基于NLRP3/Caspase-1/GSDMD焦亡通路探索小鼠急性肺损伤动态时间模型的建立[J].中国实验动物学报,2024,32(4):423-434,12.基金项目
国家自然科学基金面上项目(82174361). Funded by General Program of National Natural Science Foundation of China(82174361). （82174361）
