Figure 5.
hucMSC-Ex Induces ERK1/2 Phosphorylation and Bcl2 Expression and Inhibits the IKKB/NFkB/Caspase-9/-3 Pathway
(A) Western blot quantification of pERK1/2, total ERK1/2, Bcl2, pIKKβ, pNFkB, total NFkB, and casp9 and casp3. hucMSC-Ex induced ERK1/2 phosphorylation and Bcl2 expression and inhibited IKKB/NFkB/casp-9/-3 signaling in H2O2- or CCl4-injured L02 cells. (B) Bcl2 and pNFkB expression quantified by western blot (i). hucMSC-Ex dose-dependently inhibited pNFkB and induced Bcl2 expression in CCl4-injured L02 cells. Representative immunofluorescent images of pNFkB and Bcl2 expression in CCl4-injured L02 cells treated with PBS and hucMSC-Ex (ii). Original magnification 200×. (C) L02 cell viability was recovered by hucMSC-Ex or z-vad-FMK (n = 3). (D) CCl4-induced casp3 activity was reduced in hucMSC-Ex (20, 40, and 80 μg) (n = 3; *p < 0.05, **p < 0.01). (E) Western blot quantification of cleaved casp3 (i) in CCl4-injured L02 cells and immunohistochemical staining of cleaved casp3 in CCl4-injured mouse liver (ii). hucMSC-Ex inhibited cleaved casp3 expression of CCl4-injured hepatocytes in vitro and in vivo. Original magnification 200×. Data are expressed as relative ratios of specific proteins to GAPDH and shown as numbers under individual blots (n = 3; *p < 0.05).