4.5. RAW 264.7 Macrophage Viability Test

The MTT colorimetric method was used to detect the effect of compounds 1–10 on the viability of RAW 264.7 macrophages. The RAW 264. 7 macrophages in the logarithmic growth phase were digested with trypsin to prepare a single-cell suspension, which was seeded in a 96-well plate at a density of 1 × 10⁴ cells per well and cultured in a 5% CO₂ incubator for 24 h at 37 °C, before discarding the supernatant. The blank control group was cultured with 10% FBS-containing DMEM, and the drug group was treated with aqueous solutions of compounds 1–10, with six replicate wells for each concentration. Incubation was continued in 5% CO₂ at 37 °C. After 24 h of incubation, 10 μL of 5 mg/mL MTT was added to each well. The culture solution was removed after culturing for 4 h. Then, 100 μL of DMSO was added to each well, before shaking for 10 min to achieve complete dissolution. The optical density (OD) was measured at 492 nm using a microplate reader to calculate cell viability.