Association between ASAH1 and pyroptosis during acute lung injury in septic mice
10.3760/cma.j.cn131073.20240308.00722
- VernacularTitle:脓毒症小鼠急性肺损伤时ASAH1与细胞焦亡的关系
- Author:
Beiying WANG
1
;
Dapeng LI
;
Dandan ZHANG
;
Weiwei QIN
;
Lixin SUN
;
Wei HAN
Author Information
1. 南京医科大学青岛临床医学院，青岛 266071
- Keywords:
Sepsis;
Acute lung injury;
ASAH1;
Pyroptosis
- From:
Chinese Journal of Anesthesiology
2024;44(7):876-880
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the association between N-acylsphingosine amide hydrolase 1 (ASAH1) and pyroptosis during acute lung injury (ALI) in septic mice.Methods:Forty SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-23 g, were divided into 4 groups ( n=10 each) by a random number table method: sham operation group (Sham group), ALI group, HCFU solvent+ ALI group (HA group) and ASAH1 inhibitor HCFU+ ALI group (AA group). The abdominal cavity was only opened in Sham group, and cecal ligation puncture was performed in ALI, HA and AA groups. HCFU solvent 0.2 ml was intraperitoneally injected at 2 h before operation in HA group, and HCFU 10 mg/kg was intraperitoneally injected at 2 h before operation in AA group. The mice were sacrificed at 24 h under deep anesthesia, the eyeballs were removed to collect the blood, the bronchoalveolar lavage fluid (BALF) was collected, and lung tissues and blood samples were collected for microscopic examination of the pathological changes (using HE staining) which were scored and for determination of concentrations of protein in BALF (by BCA method), concentrations of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) in BALF (by enzyme-linked immunosorbent assay), and expression of NOD-like receptor thermoprotein structural domain-related protein 3 (NLRP3) in lung tissues (by Western blot), gasdermin D protein (GSDMD), ASAH1 and cysteine protease-1 (caspase-1) (by Western blot). The wet/dry lung weight (W/D) ratio was calculated. Results:Compared with Sham group, the lung injury score, W/D ratio and concentrations of protein in BALF, IL-1β, IL-6 and TNF-α in serum were significantly increased, and the expression of NLRP3, GSDMD and caspase-1 in lung tissues was up-regulated in ALI, HA and AA groups, and the expression of ASAH1 was significantly up-regulated in ALI and HA groups ( P<0.05). Compared with ALI and HA groups, the lung injury score, W/D ratio, and concentrations of protein in BALF, IL-1β, IL-6 and TNF-α in serum were significantly increased, the expression of NLRP3, GSDMD and caspase-1 in lung tissues was up-regulated, and the expression of ASAH1 was down-regulated in AA group ( P<0.05 or 0.01). Conclusions:ASAH1 is involved in the endogenous protective mechanism underlying ALI in septic mice, which may be related to the inhibition of cell pyroptosis.
