Effect of amphotericin B on the production of tumor necrosis factor-α and interleukin-8 and activation of signaling molecule p38MAPK in human THP-1 cells

Objective To evaluate the effect of amphotericin B on the production of tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) and activation of p38 mitogen-activated protein kinases (p38MAPK) in a human acute monocytic leukemia cell line (THP-1) . Methods Cultured THP-1 cells were divided into several groups: blank control group receiving no treatment, amphotericin B groups treated with 2, 4 and 8 mg/L amphotericin B separately, positive control group treated with 100 μg/L β-glucosan or 100 mg/L lipopolysaccharide. Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of TNF-α and IL-8 after the THP-1 cells were treated with different stimuli for some durations. Enzyme-linked immunosorbent assay (ELISA) was conducted to detect the level of TNF-α in the culture supernatant of THP-1 cells after 24-hour treatment with 8 mg/L amphotericin B, and Western blot analysis to measure the levels of p38MAPK and phosphorylated p38MAPK after 30-minute treatment with 8 mg/L amphotericin B. Results After 6-hour treatment with 2, 4 and 8 mg/L amphotericin B separately, the mRNA levels of TNF-α in THP-1 cells (7.55 ± 1.17, 19.47 ± 2.91, 57.22 ± 0.65) and IL-8 (2.98 ± 0.04, 5.22 ± 1.35, 11.82 ± 1.66) were all significantly higher than those in the blank control group (TNF-α: 1.00 ± 0.07, P < 0.01, 0.001, 0.001 respectively; IL-8: 1.01 ± 0.23, P < 0.01, 0.001, 0.001 respectively) . After the treatment with 8 mg/L amphotericin B for 1, 3, 6 hours, the mRNA levels of TNF-α (8.61 ± 0.30, 10.75 ± 0.08, 56.98 ± 2.43) and IL-8 (2.63 ± 0.28, 5.35 ± 0.98, 11.73 ± 1.18) in THP-1 cells were all significantly higher than those in the blank control group (TNF-α: 1.18 ± 0.17, P < 0.05, 0.01, 0.001; IL-8: 1.23 ± 0.11, P < 0.05, 0.01, 0.001) . After 24-hour treatment with 8 mg/L amphotericin B, the level of TNF-α in the culture supernatant of THP-1 cells was significantly higher than that in the blank control group (4 039.06 ± 223.87 ng/L vs. 96.31 ± 0.26 ng/L, P < 0.001) . Conclusion Amphotericin B can promote the p38MAPK phosphorylation and increase the levels of TNF-α and IL-8 in human THP-1 cells in vitro, suggesting its immunomodulatory effects. Key words: Amphotericin B; Tumor necrosis factor-alpha; Interleukin-8; p38 Mitogen-activated protein kinases; THP-1 monocytes