Deferoxamine reduces neuronal death and hematoma lysis after intracerebral hemorrhage in aged rats

Abstract

Intracerebral hemorrhage (ICH) is primarily a disease of the elderly. Deferoxamine (DFX), an iron chelator, reduces long-term neurological deficits and brain atrophy after ICH in aged rats. In the present study, we investigated whether DFX can reduce acute ICH-induced neuronal death and whether it affects the endogenous response to ICH (ferritin upregulation and hematoma resolution) in aged rats. Male Fischer 344 rats (18 months old) had an intracaudate injection of 100 μL autologous whole blood into the right basal ganglia and were treated with DFX (100 mg/kg) or vehicle 2 hours post-ICH and then every 12 hours up to 7 days. Rats were euthanized 1, 3, or 7 days later for neuronal death, ferritin and hematoma size measurements. Plasma ferritin levels and behavioral outcome following ICH were also examined. DFX treatment significantly reduced ICH-induced neuronal death and neurological deficits. DFX also suppressed ferritin upregulation in the ipsilateral basal ganglia after ICH and hematoma lysis (hematoma volume at day 7: 13.2±4.9 vs. 3.8±1.2 mm3 in vehicle-treated group, p<0.01). However, effects of DFX on plasma ferritin levels after ICH did not reach significance. In conclusion, DFX reduces neuronal death and neurological deficits after ICH in aged rats. It also affects the endogenous response to ICH.

Keywords: Cerebral hemorrhage; deferoxamine; iron; neuronal death.

Conflict of interest statement

Conflict of Interest:

Tetsuhiro Hatakeyama, Masanobu Okauchi, Ya Hua, Richard F. Keep and Guohua Xi declare that they have no conflict of interest.

Figures

Figure 1

A) Fluoro-Jade C positive cells in the ipsilateral basal ganglia 1 day after ICH in vehicle- and DFX-treated groups. B) Fluoro-Jade C positive cell counts in the ipsilateral basal ganglia 1 and 3 days after ICH in vehicle- and DFX-treated groups. Values are means±SD, n=5 rats per group, *P<0.05 vs. ICH+vehicle group. Scale bar = 200 μm.

Figure 2

A) TUNEL positive cells in the ipsilateral basal ganglia 1 day after ICH in vehicle- and DFX-treated groups. B) TUNEL positive cell counting in the ipsilateral basal ganglia 1 and 3 days after ICH in vehicle- and DFX-treated groups. Values are means±SD, n=5 rats per group, * P<0.05 vs. ICH+vehicle group. Scale bar = 200 μm.

Figure 3

Forelimb-placing (A) and corner turn (B) test scores before ICH (pretest) and 1, 3 and 7 days after ICH (ND = not detectable). Values are means±SD. n=9 rats per group, *P<0.05 vs. ICH+vehicle group, **P<0.01 vs. ICH+vehicle group.

Figure 4

A) Coronal sections (hematoxylin and eosin staining) from brains 7 days after ICH in vehicle and DFX-treated groups. B) Bar graph showing hematoma volume 1, 3 and 7 days after ICH in vehicle- and DFX-treated groups. Values are means±SD. n=4 rats per group, **P<0.01 vs. ICH+vehicle group. Scale bar = 5.0 mm.

Figure 5

A) Western blot analysis showing the time course of ferritin heavy-chain (FTH) levels in the ipsilateral basal ganglia 1, 3 and 7 days after ICH in vehicle-treated group. Bar graph quantifies the Western blotting. Beta-actin is a loading control. B) Western blot analysis showing the time course of the levels of ferritin light-chain (FTL) levels in the ipsilateral basal ganglia 1, 3 and 7 days after ICH in vehicle-treated group. Bar graph quantifies the Western blotting. Values are mean±SD, n=3 rats per group, *P<0.05 vs. ICH+vehicle group, **P<0.01 vs. ICH+vehicle group.

Figure 6

A) Ferritin immunoreactivity in the ipsilateral basal ganglia 7 days after ICH in vehicle and DFX-treated rats. B) Western blot analysis showing ferritin heavy-chain levels in the ipsilateral basal ganglia 7 days after ICH in vehicle and DFX-treated groups. Bar graph quantifies the Western blotting. C) Western blot analysis showing the levels of ferritin light-chain in the ipsilateral basal ganglia 7 days after ICH in vehicle and DFX-treated groups. Bar graph quantifies the Western blotting. Values are means±SD, n=3 rats per group, *P<0.05 vs. ICH+vehicle group. Scale bar = 200 μm.