PMID- 32173407 OWN - NLM STAT- MEDLINE DCOM- 20210205 LR - 20211229 IS - 1879-0720 (Electronic) IS - 0928-0987 (Linking) VI - 147 DP - 2020 Apr 30 TI - Investigation on the mechanism of mafenide in inhibiting pyroptosis and the release of inflammatory factors. PG - 105303 LID - S0928-0987(20)30092-0 [pii] LID - 10.1016/j.ejps.2020.105303 [doi] AB - OBJECTIVE: The present study was designed to investigate the roles and mechanism of mafenide (MAF) in targeted inhibition of Gasdermin D (GSDMD) cleavage and in suppressing pyroptosis. METHODS: Lipopolysaccharide (LPS) and Nigericin were used to induce pyroptosis in mouse bone marrow-derived macrophages (iBMDM) and mouse microglia (BV2). Lactate dehydrogenase (LDH) release rate and Propidium Iodide (PI) uptake rate were used to detect cytotoxicity, Western blot was used to detect the protein expression, and Enzyme-linked immunosorbent assay (ELISA) was utilized to detect the expression of inflammatory factors from culture medium. MAF was labeled with biotin and subsequently subjected to Pull-down assay to detect its binding to GSDMD. GSDMD-Asp275 site was further mutated to validate the binding of MAF to GSDMD. Finally, the effects of MAF on inflammatory factor release and microglial activation were confirmed in the APP/PS12 animal model. RESULTS: MAF could inhibit pyroptosis in iBMDM and microglia BV2, and decrease the release of inflammatory factors. MAF could inhibit GSDMD cleavage by directly binding to the GSDMD-Asp275 site, while the expression of p30-GSDMD was simultaneously down-regulated and the release of inflammatory factors was decreased. MAF could reduce the levels of inflammatory factors in cerebrospinal fluid and peripheral blood of APP/PS1 mice, and suppress the activation of microglia. CONCLUSION: The mechanism underlying the regulation of MAF on inflammatory response was correlated with the inhibition of pyroptosis. MAF could inhibit GSDMD cleavage by directly binding to GSDMD. CI - Copyright (c) 2020. Published by Elsevier B.V. FAU - Han, Chenyang AU - Han C AD - State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, 210093, China; Department of pharmacy, The Second Affiliated Hospital of Jiaxing University, Zhejiang 314001, China. Electronic address: 691513770@qq.com. FAU - Yang, Yi AU - Yang Y AD - Department of pharmacy, The Second Affiliated Hospital of Jiaxing University, Zhejiang 314001, China. Electronic address: wasd911@126.com. FAU - Yu, Anqi AU - Yu A AD - State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing 210093, China. Electronic address: 1144900357@qq.com. FAU - Guo, Li AU - Guo L AD - Department of Center Laboratory, The Second Affiliated Hospital of Jiaxing University, Zhejiang 314001, China. Electronic address: taishanlg@126.com. FAU - Guan, Qiaobing AU - Guan Q AD - Department of neurology, The Second Affiliated Hospital of Jiaxing University, Zhejiang 314001, China. Electronic address: guanqb@126.com. FAU - Shen, Heping AU - Shen H AD - Department of neurology, The Second Affiliated Hospital of Jiaxing University, Zhejiang 314001, China. Electronic address: shenhp_77@163.com. FAU - Jiao, Qingcai AU - Jiao Q AD - State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing, 210093, China. Electronic address: jiaoqc@163.com. LA - eng PT - Journal Article DEP - 20200312 PL - Netherlands TA - Eur J Pharm Sci JT - European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences JID - 9317982 RN - 0 (Cytokines) RN - 0 (Gsdmd protein, mouse) RN - 0 (Intracellular Signaling Peptides and Proteins) RN - 0 (Lipopolysaccharides) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (Nlrp3 protein, mouse) RN - 0 (Phosphate-Binding Proteins) RN - 58447S8P4L (Mafenide) RN - EC 3.4.22.36 (Casp1 protein, mouse) RN - EC 3.4.22.36 (Caspase 1) RN - RRU6GY95IS (Nigericin) SB - IM EIN - Eur J Pharm Sci. 2022 Mar 1;170:106099. PMID: 34963568 MH - Animals MH - Caspase 1/metabolism MH - Cell Culture Techniques MH - Cell Death/drug effects MH - Cytokines/*metabolism MH - Intracellular Signaling Peptides and Proteins/metabolism MH - Lipopolysaccharides/pharmacology MH - Macrophages/drug effects MH - Mafenide/*pharmacology MH - Male MH - Mice MH - Microglia/drug effects MH - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism MH - Nigericin/pharmacology MH - Phosphate-Binding Proteins/metabolism MH - Pyroptosis/*drug effects OTO - NOTNLM OT - Gasdermin D OT - Inflammatory response OT - Mafenide OT - Pyroptosis COIS- Declaration of Competing Interest We have no conflicts of interest to declare. EDAT- 2020/03/17 06:00 MHDA- 2021/02/07 06:00 CRDT- 2020/03/17 06:00 PHST- 2019/11/26 00:00 [received] PHST- 2020/02/19 00:00 [revised] PHST- 2020/03/08 00:00 [accepted] PHST- 2020/03/17 06:00 [pubmed] PHST- 2021/02/07 06:00 [medline] PHST- 2020/03/17 06:00 [entrez] AID - S0928-0987(20)30092-0 [pii] AID - 10.1016/j.ejps.2020.105303 [doi] PST - ppublish SO - Eur J Pharm Sci. 2020 Apr 30;147:105303. doi: 10.1016/j.ejps.2020.105303. Epub 2020 Mar 12.