Free radical scavenger (edaravone) prevents endotoxin-induced liver injury after partial hepatectomy in rats

Experimental protocol. Rats were treated with lipopolysaccharide (LPS) (1.5mg/kg) 48h after 70% hepatectomy. Edaravone (10mg/kg) was administered by a single intravenous injection 0.5h before LPS treatment.

Effect of edaravone on survival of rats. (A) Dose dependency: rats were treated with LPS (1.5mg/kg) 48h after partial hepatectomy. Edaravone (1–10mg/kg) was administered 0.5h before LPS injection. (10 rats/group) (B) Optimal point of edaravone injection for maximum effect on survival: rats were treated with LPS as described above. Edaravone (10mg/kg) was administered at 0.5h before, immediately after, or 0.5h after LPS injection. Data represent percentage of animals surviving (5 rats/group). *P<0.05, **P<0.01 vs. vehicle-treated rats with LPS.

Effect of edaravone on serum AST and LDH. Partially hepatectomized rats with LPS were treated with, or without, edaravone. Blood samples were collected at the indicated times after LPS treatment. (A) AST and (B) LDH. Data represent mean±SD (n=5–10 rats). *P<0.05 vs. vehicle-treated rats with LPS.

Effect of edaravone on pathological change of the liver, spleen and kidney. Liver sections were obtained from LPS-untreated rats 48h after partial hepatectomy for the untreated (A) and the edaravone-treated (B) groups. Liver, spleen, and kidney sections from partially hepatectomized rats with LPS; in the edaravone-untreated (C, E, G) and the treated (D, F, H) (hematoxylin–eosin staining, original magnification×100).

Effect of edaravone on serum levels of MDA. Partially hepatectomized rats with LPS were treated with, or without, edaravone. Blood samples were collected at the indicated times after LPS treatment. Data represent mean±SD (n=5–10). *P<0.05 vs. vehicle-treated rats with LPS.

Effect of edaravone on serum levels of TNF-a, IL-1β, IL-12, IFN-γ, and CINC-1. Partially hepatectomized rats with LPS were treated with, or without, edaravone. Partially hepatectomized rats without LPS were treated with, or without, edaravone. Blood samples were collected at the indicated times after LPS treatment. (A) TNF-α, (B) IL-1β, (C) IL-12, (D) IFN-γ, and (E) CINC-1. Data represent mean±SD (n=5–10). *P<0.05 vs. vehicle-treated rats with LPS.

Effect of edaravone on production of nitric oxide. Rats were treated with LPS (1.5mg/kg) 48h after partial hepatectomy. Edaravone (10mg/kg) was administered by a single intravenous injection 0.5h before LPS treatment. Determination of nitric oxide production (nitrite and nitrate). Blood samples were obtained at the indicated times after LPS treatment with, or without, edaravone. Data represent mean±SD (n=5). *P<0.05 vs. vehicle-treated rats with LPS.

Effect of edaravone on levels of TNF-α, IL-1β, IL-12, IFN-γ, and CINC-1 in the liver. Partially hepatectomized rats with LPS were treated with, or without, edaravone. Liver samples were collected at the indicated times after LPS treatment. (A) TNF-α, (B) IL-1β, (C) IL-12, (D) IFN-γ, and (E) CINC-1. Data represent mean±SD (n=5–10). *P<0.05 vs. vehicle-treated rats with LPS.

Effect of edaravone on NF-κB activation. Rats were treated with LPS (1.5mg/kg) 48h after partial hepatectomy. Edaravone (10mg/kg) was administered by a single intravenous injection 0.5h before LPS treatment. (A) Nuclear extracts (4μg of protein) were prepared from liver at the indicated times, and NF-κB was analyzed by electrophoretic mobility shift assay was performed. (B) The bands corresponding to NF-κB were quantitated by densitometry. Data represent mean±SD (n=5). *P<0.05 vs. vehicle-treated rats with LPS.

Effect of edaravone on induction of iNOS. Liver samples were obtained at the indicated times. (A) Northern blot analysis of mRNA levels for iNOS and GAPDH in entire livers. Total RNA (10μg) was loaded onto each lane. (B) Western blot analysis of iNOS protein in entire livers. The cell lysate (260μg protein) was subjected to SDS–PAGE. Samples shown are representative of five samples from each group and time point.