Thp 1 cell

The THP-1 cell used to construct the immune infiltration model, The THP-1 cell was purchased from Wuhan (Procell, China), used for in vitro research and cultivated in RPMI-1640 medium containing 10% foetal bovine serum (Gibco, MA, United States) and 0.05 mM β- Mercaptoethanol in a 37°C cell incubator with 5% CO₂. Prior to each experiment, 1.5*10⁶/mL THP-1 cells were inoculated in six-well plates and treated with 100 ng/mL Phorbol-12-myristate-13-acetate (PMA) for 24 h to induce differentiation into macrophages. PMA-induced THP-1 cells were treated as described below. Normal control group (NC): PMA-induced THP-1 cells were cultured in RPMI-1640 medium for 48 h. LPS + INF-Y group: PMA-induced THP-1 cells were incubated with 100 ng/mL lipopolysaccharide (LPS) and 20 ng/mL IFN-γ for 48 h. LPS + INF-Y + Resveratrol: THP-1 cell was first incubated with 2.5 uM Resveratrol for 2 h. LPS + INF-Y+ magnolol: HL-1 was first incubated with 15 uM magnolol for 2 h.

Human monocyte derived from patients with acute monocytic leukemia, specifically THP-1 cell (Procell, Wuhan, China), was used in the study. The chemicals utilized in this study, along with the corresponding companies from which they were purchased, are listed in Supplementary Table S1. The differentiation from THP-1 cell to adherent macrophage was induced by an incubation with 50 nM phorbol 12-myristate 13-acetate (PMA, REF: P8139; Sigma, United States) for 24 h. To synchronize the cells, the differentiated macrophages were cultured in basal RPMI 1640 medium for 24 h and then allocated into the following groups: control, oxLDL, oxLDL + L-Cth 0.1 mM, oxLDL + L-Cth 0.3 mM, and oxLDL + L-Cth 1.0 mM. oxLDL and L-Cth were procured from Zhongshan Golden Bridge and Sigma-Aldrich, respectively. In the oxLDL group, 50 mg/L oxLDL was added and incubated for 6 h. In the oxLDL + L-Cth 0.1 mM group, oxLDL + L-Cth 0.3 mM group, and oxLDL + L-Cth 1.0 mM group, the cells were pre-treated with L-Cth for 30 min and subsequently exposed to 50 mg/L oxLDL for 6 h (Zhu et al., 2014 (link)). Likewise, in the oxLDL + aminooxy acetic acid (AOAA; REF: S4989; Selleck, United States) group and the oxLDL + S-adenosylmethionine (SAM; REF: S910367; Macklin, China) group, the cells were intervened with AOAA or SAM for 30 min, followed by a treatment with 50 mg/L oxLDL for 6 h.

Human monocytic leukemia THP-1 cell was purchased from Procell Life Science & Technology Co., Ltd (Wuhan, Hubei Province, China). Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Gibco, USA), 100  U/mL penicillin, and 100 μg/mL streptomycin (Gibco, USA). They were maintained in an incubator at 37°C with 5% CO₂ and saturated humidity, and the culture medium was replaced with complete culture medium every 2 or 3 days. The cells were treated with phorbol-12-myristate-13-acetate (PMA) (100 nM) for 3 h to induce their differentiation into resting M0 macrophage. A fresh complete culture medium was added and cultured for 24 h after washing the cells with phosphate-buffered saline (PBS). Differentiated THP-1 cells were then stimulated with MSU (50, 100, and 200 μg/mL) with optimal concentration and treated with TFDC at final concentrations of 25, 50, 75, 100, and 125 μg/mL as well as colchicine (positive drug, 2 μg/mL) simultaneously at 37°C for 24 h [33 ].