The effect of GSDMD gene knockout on lung injury and ROS/NLRP3/Caspase-1 pathway of pneumonia mice_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

 YUAN Meng , DONG Huiwen , LIU Jiali , FENG Xuehui , HAN Baohua ,  HAN Shuchi , ZHANG Zhibin

Department of emergency The First Affiliated Hospital of Hebei North University, zhangjiakou, Hebei 075000, P.R. China;

Corresponding?author：

HAN Shuchi, Email: zjkhanshuchi@sina.cn

Keywords：

Pneumonia; lung injury; GSDMD; gene knockout; pyroptosis

DOI：

10.7507/1671-6205.202503090

Video：

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Objective To investigate the effect of Gasdermin D(GSDMD) gene knockout on lung injury and reactive oxygen species (ROS)/nod-like receptor protein 3 (NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1) pyroptosis pathway of pneumonia mice. Methods 20 wild-type (WT) male C57BL/6J mice were randomly divided into WT control group and WT model group. 20 GSDMD gene knockout (KO) mice were randomly divided into KO control group and KO model group, each group consisted of 10 animals. The pneumonia model was induced by puncturing the nasal mucosa with a sterile needle and slowly instilling Streptococcus pneumoniae suspension in WT model group and KO model group. Serum and bronchoalveolar lavage fluid (BALF) were collected 7 days after modeling, and the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected; the lung tissues were collected and the dry wet ratio (W/D), pathological changes, the levels of malondialdehyde (MDA), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), ROS activity, and the protein expression levels of GSDMD, GSDMD-N, NLRP3, Caspase-1 were detected. Results The lung tissue of WT model group showed cell infiltration with alveolar septal thickening, the ratio of W/d in lung tissue, the levels of IL-1β, IL-6 and TNF-α in serum and BALF, the levels of MDA, ROS, NLRP3 and Caspase-1 in lung tissue were all higher than those in WT control group, the levels of T-AOC and SOD in lung tissue were lower than those in WT control group (P<0.05), and the levels of serum, BALF and lung tissue in KO control group were not significantly different from those in WT control group (P>0.05) There was no expression of GSDMD-N in the lung tissue of KO model group, lung tissue w/d ratio, serum and BALF levels of IL-1β, IL-6, TNF-α, MDA, ROS activity, GSDMD-N, NLRP3 and Caspase-1 protein expression were lower than those in WT model group, the levels of T-AOC and SOD in lung tissue were higher than those in WT Model Group (P<0.05). Conclusion GSDMD gene knockout attenuates lung injury and inhibits ROS/NLRP3/Caspase-1 pathway in pneumonia mice.

Citation： YUAN Meng, DONG Huiwen, LIU Jiali, FENG Xuehui, HAN Baohua, HAN Shuchi, ZHANG Zhibin. The effect of GSDMD gene knockout on lung injury and ROS/NLRP3/Caspase-1 pathway of pneumonia mice. Chinese Journal of Respiratory and Critical Care Medicine, 2025, 24(6): 407-412. doi: 10.7507/1671-6205.202503090 Copy

1.	Rajbanshi A, Giri P, Laha S, et al. Epidemiology, clinical presentation, and respiratory sequelae of severe adenoviral pneumonia in children admitted in a tertiary care pediatric intensive care unit from Eastern India: A single-center observational study. Journal of Pediatric Critical Care, 2022, 9(4): 131-138.
2.	Ji K, Sun J, Yan Y, et al. Epidemiologic and clinical characteristics of human bocavirus infection in infants and young children suffering with community acquired pneumonia in Ningxia, China. Virol J, 2021, 18(1): 212.
3.	Li L, Dai B, Sun Y, et al. The activation of IL-17 signaling pathway promotes pyroptosis in pneumonia-induced sepsis. Ann Transl Med, 2020, 8(11): 674.
4.	Wang X, Zhao Y, Wang D, et al. ALK-JNK signaling promotes NLRP3 inflammasome activation and pyroptosis via NEK7 during Streptococcus pneumoniae infection. Mol Immunol, 2023, 157: 78-90.
5.	帥維正, 李軍, 陳旭昕, 等. Gasdermin D蛋白對內毒素導致肺泡巨噬細胞焦亡的影響. 中華肺部疾病雜志, 2020, 13(4): 441-445.
6.	Qu R, Liu J, Feng L, et al. Down-regulation of KLF9 ameliorates LPS-caused acute lung injury and inflammation in mice via reducing GSDMD expression. Autoimmunity, 2022, 55(8): 11-10.
7.	帥維正, 李軍, 李大偉, 等. GSDMD基因敲除對內毒素所致小鼠急性肺損傷的保護作用. 解放軍醫學雜志, 2020, 45(11): 1125-1130.
8.	王亞靜, 張靜, 宋衛衛, 等. 大黃素通過調控ROS/TXNIP/NLRP3軸介導的細胞焦亡途徑減輕重癥肺炎大鼠肺損傷. 中國病理生理雜志, 2021, 37(7): 1240-1245.
9.	Lenga M, Fournet M, Zhai R, et al. Receptor for Advanced Glycation End-Products Promotes Activation of Alveolar Macrophages through the NLRP3 Inflammasome/TXNIP Axis in Acute Lung Injury. Int J Mol Sci, 2022, 23(19): 11659.
10.	Soltanieh A, Avizeh R, Najafzadeh V, et al. Antioxidant effect of montelukast on acute lung injury induced by lipopolysaccharide in dogs. J Babol Univ Med Sci, 2021, 23(1): 229-235.
11.	Zhan P, Lu X, Li Z, et al. Mitoquinone alleviates bleomycin-induced acute lung injury via inhibiting mitochondrial ROS-dependent pulmonary epithelial ferroptosis. Int Immunopharmacol, 2022, 113: 109359.
12.	張杰, 葉慧芳, 單艷華, 等. 硫化氫對脂多糖誘發的大鼠急性肺損傷肺組織中細胞凋亡及SOD、MDA水平的影響. 實用預防醫學, 2018, 25(6): 753-756.
13.	Xue Y, Zhang Y, Chen L, et al. Citrulline protects against LPS-induced acute lung injury by inhibiting ROS/NLRP3-dependent pyroptosis and apoptosis via the Nrf2 signaling pathway. Experimental and therapeutic medicine, 2022, 24(4): 632.
14.	Ren F, Xu J, Zhang J, et al. PM2. 5 induced lung injury through upregulating ROS-Dependent NLRP3 Inflammasome-Mediated Pyroptosis. Immunobiology, 2022, 227(3): 152207.
15.	Yi J, Li L, Yin ZJ, et al. Polypeptide from Moschus Suppresses Lipopolysaccharide-Induced Inflammation by Inhibiting NF-kappa B-ROS/NLRP3 Pathway. Chin J Integr Med, 2023, 29(10): 895-904.
16.	Tao H, Zhao H, Mo A, et al. VX-765 attenuates silica-induced lung inflammatory injury and fibrosis by modulating alveolar macrophages pyroptosis in mice. Ecotoxicology and Environmental Safety, 2023, 249: 114359.
17.	Lin F, Xiao J, Liang F, et al. Preconditioning with rHMGB1 ameliorates lung ischemia-reperfusion injury by inhibiting alveolar macrophage pyroptosis via the Keap1/Nrf2/HO-1 signaling pathway. Journal of translational medicine, 18(1): 301.
18.	Huang W, Chen X, Yin M, et al. Protection effects of mice liver and lung injury induced by coronavirus infection of Qingfei Paidu decoction involve inhibition of the NLRP3 signaling pathway. J Ethnopharmacol, 2024, 1(321): 117512.
19.	Chen Y, Jiang Y, Liu X, et al. Polydatin alleviates mycoplasma pneumoniae-induced injury via inhibition of Caspase-1/GSDMD-dependent pyroptosis. Int J Med Microbiol, 2023, 313(5): 151586.
20.	Song Z, Han C, Luo G, et al. Yinqin Qingfei granules alleviate Mycoplasma pneumoniae pneumonia via inhibiting NLRP3 inflammasome-mediated macrophage pyroptosis. Front Pharmacol, 2024, 27(15): 1437475.

1. Rajbanshi A, Giri P, Laha S, et al. Epidemiology, clinical presentation, and respiratory sequelae of severe adenoviral pneumonia in children admitted in a tertiary care pediatric intensive care unit from Eastern India: A single-center observational study. Journal of Pediatric Critical Care, 2022, 9(4): 131-138.
2. Ji K, Sun J, Yan Y, et al. Epidemiologic and clinical characteristics of human bocavirus infection in infants and young children suffering with community acquired pneumonia in Ningxia, China. Virol J, 2021, 18(1): 212.
3. Li L, Dai B, Sun Y, et al. The activation of IL-17 signaling pathway promotes pyroptosis in pneumonia-induced sepsis. Ann Transl Med, 2020, 8(11): 674.
4. Wang X, Zhao Y, Wang D, et al. ALK-JNK signaling promotes NLRP3 inflammasome activation and pyroptosis via NEK7 during Streptococcus pneumoniae infection. Mol Immunol, 2023, 157: 78-90.
5. 帥維正, 李軍, 陳旭昕, 等. Gasdermin D蛋白對內毒素導致肺泡巨噬細胞焦亡的影響. 中華肺部疾病雜志, 2020, 13(4): 441-445.
6. Qu R, Liu J, Feng L, et al. Down-regulation of KLF9 ameliorates LPS-caused acute lung injury and inflammation in mice via reducing GSDMD expression. Autoimmunity, 2022, 55(8): 11-10.
7. 帥維正, 李軍, 李大偉, 等. GSDMD基因敲除對內毒素所致小鼠急性肺損傷的保護作用. 解放軍醫學雜志, 2020, 45(11): 1125-1130.
8. 王亞靜, 張靜, 宋衛衛, 等. 大黃素通過調控ROS/TXNIP/NLRP3軸介導的細胞焦亡途徑減輕重癥肺炎大鼠肺損傷. 中國病理生理雜志, 2021, 37(7): 1240-1245.
9. Lenga M, Fournet M, Zhai R, et al. Receptor for Advanced Glycation End-Products Promotes Activation of Alveolar Macrophages through the NLRP3 Inflammasome/TXNIP Axis in Acute Lung Injury. Int J Mol Sci, 2022, 23(19): 11659.
10. Soltanieh A, Avizeh R, Najafzadeh V, et al. Antioxidant effect of montelukast on acute lung injury induced by lipopolysaccharide in dogs. J Babol Univ Med Sci, 2021, 23(1): 229-235.
11. Zhan P, Lu X, Li Z, et al. Mitoquinone alleviates bleomycin-induced acute lung injury via inhibiting mitochondrial ROS-dependent pulmonary epithelial ferroptosis. Int Immunopharmacol, 2022, 113: 109359.
12. 張杰, 葉慧芳, 單艷華, 等. 硫化氫對脂多糖誘發的大鼠急性肺損傷肺組織中細胞凋亡及SOD、MDA水平的影響. 實用預防醫學, 2018, 25(6): 753-756.
13. Xue Y, Zhang Y, Chen L, et al. Citrulline protects against LPS-induced acute lung injury by inhibiting ROS/NLRP3-dependent pyroptosis and apoptosis via the Nrf2 signaling pathway. Experimental and therapeutic medicine, 2022, 24(4): 632.
14. Ren F, Xu J, Zhang J, et al. PM2. 5 induced lung injury through upregulating ROS-Dependent NLRP3 Inflammasome-Mediated Pyroptosis. Immunobiology, 2022, 227(3): 152207.
15. Yi J, Li L, Yin ZJ, et al. Polypeptide from Moschus Suppresses Lipopolysaccharide-Induced Inflammation by Inhibiting NF-kappa B-ROS/NLRP3 Pathway. Chin J Integr Med, 2023, 29(10): 895-904.
16. Tao H, Zhao H, Mo A, et al. VX-765 attenuates silica-induced lung inflammatory injury and fibrosis by modulating alveolar macrophages pyroptosis in mice. Ecotoxicology and Environmental Safety, 2023, 249: 114359.
17. Lin F, Xiao J, Liang F, et al. Preconditioning with rHMGB1 ameliorates lung ischemia-reperfusion injury by inhibiting alveolar macrophage pyroptosis via the Keap1/Nrf2/HO-1 signaling pathway. Journal of translational medicine, 18(1): 301.
18. Huang W, Chen X, Yin M, et al. Protection effects of mice liver and lung injury induced by coronavirus infection of Qingfei Paidu decoction involve inhibition of the NLRP3 signaling pathway. J Ethnopharmacol, 2024, 1(321): 117512.
19. Chen Y, Jiang Y, Liu X, et al. Polydatin alleviates mycoplasma pneumoniae-induced injury via inhibition of Caspase-1/GSDMD-dependent pyroptosis. Int J Med Microbiol, 2023, 313(5): 151586.
20. Song Z, Han C, Luo G, et al. Yinqin Qingfei granules alleviate Mycoplasma pneumoniae pneumonia via inhibiting NLRP3 inflammasome-mediated macrophage pyroptosis. Front Pharmacol, 2024, 27(15): 1437475.

Chinese Journal of Respiratory and Critical Care Medicine

The effect of GSDMD gene knockout on lung injury and ROS/NLRP3/Caspase-1 pathway of pneumonia mice

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