The expression of IL-8 mRNA in human neutrophils following exposure to hemin.

Neutrophils (5 × 10⁶ cells) were incubated for 30 minutes at 37°C with 3 μM hemin in the presence (H3 + HSA) or absence (H3) of 1% human serum albumin, or in the presence of 10 nM BIM (H3 + BIM). Control groups received only PBS (control) or albumin (HSA) as treatment. For positive IL-8 transcript production, cells were treated with 10 ng/mL LPS. After that, cells were washed 2 times with PBS and incubated for 4 hours in RPMI. Total cellular RNA was isolated, and RT-PCR was performed using IL-8 and GAPDH-specific probes in the presence of α[³²P]-CTP. Data are presented as the ratio between the expression of IL-8 to GAPDH levels. For additional information, see “Materials and methods.” This representative experiment was repeated 3 times, showing similar results. Data are reported as cumulative data in the form of mean ± SD. *P < .01.