Fig. 2
BMSCs increase HO-1 expression and activity, and Nrf2 expression in liver. a Quantitative reverse transcription PCR showing BMSC infusion or hemin increases expression of HO-1 relative to the ALF group (n = 6 per group, p < 0.05). Znpp did not decrease HO-1 mRNA levels. b Western blot shows that BMSCs infusion or hemin treatment significantly increases HO-1 protein (p < 0.05), but Znpp does not decrease HO-1 protein levels. Densitometry used to calculate normalized protein ratio (vs. β-actin standard). c Immunohistochemical stain (brown) of HO-1 in livers 72 h after D-Gal/LPS treatment (magnification × 200). BMSCs infusion or hemin treatment significantly increased HO-1 staining; Znpp did not decrease HO-1 staining. d HO-1 enzyme activity in livers 72 h after D-Gal/LPS treatment (n = 6 per group). HO-1 activity was significantly increased by BMSCs (p < 0.05); Znpp decreased HO-1 activity (p < 0.05). Hemin treatment also increased liver HO-1 activity (p < 0.05). e Western blot results showing that BMSCs increase Nrf2 expression of Nrf2 relative to the ALF group (p < 0.05); Znpp decreased Nrf2 expression (p < 0.05). Hemin treatment also increased Nrf2 protein levels (p < 0.05). Densitometry used to normalize protein levels (β-actin standard). Treatment groups: control, ALF, ALF followed by intravenous MSCs (ALF + MSC) 1 h post-induction, ALF followed by MSCs and Znpp (ALF + MSC + Znpp) 1 h post-induction, and ALF followed by hemin (ALF + hemin) 1 h post-induction. Data are mean ± SD. (* p < 0.05 vs. control group; $ p < 0.05 vs. ALF group; # p < 0.05 vs. ALF + MSC group). Abbreviations: ALF acute liver failure, BMSCs bone marrow-derived mesenchymal stem cells, D-Gal d-Galactosamine, HO-1 heme oxygenase-1, LPS lipopolysaccharide, Nrf2 nuclear factor-erythroid 2 p45-related factor 2, Znpp zinc protoporphyrin