Figure 8
From: Hemoglobin induces inflammation after preterm intraventricular hemorrhage by methemoglobin formation
Fenton reaction–induced TLR-4, TNFα, IL-1β and HO-1 mRNA expression in astrocyte cell cultures. Primary rabbit astrocyte cell cultures were exposed to a mixture of (NH4)Fe(SO4)2, hydrogen peroxide and ascorbate (the Fenton reaction) for four hours at concentrations of 10 μM (NH4)Fe(SO4)2 + 100 μM ascorbate + 20 μM H2O2 (white bars) or 50 μM (NH4)Fe(SO4)2, 500 μM ascorbate + 100 μM H2O2 (shaded bars). mRNA expression of TLR-4 (A), TNFα (B), IL-1β (C) and HO-1 (D) was determined using real-time PCR, as described in the Methods section. The mRNA expression of TLR-4, TNFα, IL-1β and HO-1 was normalized against GAPDH and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures and control conditions were analyzed using Mann–Whitney U. *** P <0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HO-1, heme oxygenase; SEM, standard error of the mean; TLR-4, Toll-like receptor-4.