Ml264

Lipopolysaccharides (Escherichia coli O111:B4, L2630, for animal experiments; Escherichia coli O111:B4, L4391, for cell experiments) were purchased from Merck (New Jersey, USA). ACSS2 inhibitor (cat no: S8588) was purchased from Selleck (Houston, USA). ML264 (cat no: HY-19994) was purchased from MedChemExpress (New Jersey, USA). Lotus Tetragonolobus Lectin (LTL) (cat no: FL-1321-2) was purchased from Thermo Fisher (MA, USA).
The following antibodies were used. ACSS2 antibody (cat no: ab133664) from Abcam (Cambridge, UK); KIM-1 antibody (cat no: sc-518008) from Santa Cruz (California, USA); F4/80 antibody (cat no: 28463-1-AP) from Proteintech (Wuhan, China); Cleaved GSDMD (N Terminal) antibody (cat no: A22523) and NLRP3 antibody (cat no: A5652) from Abclonal (Wuhan, China); caspase-1 antibody (cat no: 24232), anti-IL-1β antibody (cat no: 12242) from Cell Signaling Technology (Massachusetts, USA); GSDMD antibody (cat no: AF4012), NF-κB p65 antibody (cat no: AF5006), phospho-NF-κB p65 (Ser536) antibody (cat no: AF2006), and KLF5 antibody (cat no: AF7542) from Affinity (Changzhou, China).

2-, 5-, and 8-month-old male APPswe/PS1dE9 (APP/PS1) mice (n = 6 for each age group) and male C57BL/6 mice with the corresponding ages (n = 6 for each group) were purchased from Nanjing Biomedical Research Institute of Nanjing University (Nanjing, China). After a week’s balance, the mice were euthanized. Serum samples and brain tissues were collected for further analyses. In a separate experiment, 5-month-old male APP/PS1 mice (n = 12) and 5-month-old male C57BL/6 mice (n = 6) were used for ML264 administration. APP/PS1 mice were further divided randomly into two groups, namely, the AD control group (n = 6) and the ML264-treatment group (n = 6). The mice were injected intraperitoneally with 5 mg/kg ML264 (MedChemExpress) or vehicle (PBS) every other day for 15 weeks. After 15 weeks of treatment, the mice were subjected to behavioral tests. At the end of the test, all the animals were sacrificed, and the brain tissues were collected for further analyses. All the mice were housed in a specific pathogen-free room under a 12-h/12-h light/dark cycle and given free access to diet. All the animal procedures were performed in accordance with the criteria outlined in the Guide for the Care and Use of Laboratory Animals (National Institutes of Health, Bethesda, MD) and approved by the Animal Care and Use Committee of Xuanwu Hospital of Capital Medical University.