Vasculotoxic and Proinflammatory Effects of Plasma Heme: Cell Signaling and Cytoprotective Responses
Figure 4
Cytoprotective pathways are activated in sickle mice overexpressing wild type (wt)-HO-1. Nuclear extracts were isolated from livers and 30 μg of nuclear extract protein from each liver was run on a western blot and immunostained for phospho- and total p38 and Akt, and NF-κB p65. NF-κB p65 protein bands at 65 kDa were quantified by densitometry. Nuclear phospho-p38 MAPK (a) and phospho-Akt (b) were increased in mice 8 weeks after hydrodynamic infusion of Sleeping Beauty (SB)-wt-HO-1 DNA or after intraperitoneal injection of hemin chloride for 3 consecutive days, but not in control mice, given a hydrodynamic infusion of lactated Ringer’s solution (LRS) or SB-nonsense (ns)-HO-1 DNA. Total nuclear p38 MAPK (a) and Akt (b) were not different between treatment groups. NF-κB p65 was decreased in liver nuclear extracts of sickle mice injected with SB-wt-HO-1 or hemin, but not in control, LRS, or SB-ns-HO-1 treated mice (c). Serum levels of sVCAM-1 were measured by ELISA. Serum sVCAM-1 was lower in sickle mice injected with SB-wt-HO-1 DNA () or hemin (), but not in mice infused with SB-ns-HO-1 DNA when compared to mice infused with LRS (d). Values are means ± SEM; –4 mice per treatment group; compared to LRS controls using one-way ANOVA. Figure derived from [62].
