Fig 1.
Effects of calcitriol on body weight, liver weight, and liver index in DEN-induced liver fibrosis.
(A) Body weight, (B) liver weight, and (C) liver index in the CON, DEN, DEN + LVD₃, and DEN + HVD₃ groups after 8 weeks. All DEN-treated groups showed significantly lower body weight, liver weight, and liver index compared with CON, with no differences among DEN, DEN + LVD₃, and DEN + HVD₃. Data are presented as mean ± SD, n = 6/group. *p < 0.05 compared with the CON group.
Fig 2.
Effects of calcitriol on serum ALT, AST, and hepatic MDA levels in DEN-induced liver fibrosis.
(A) ALT, (B) AST, and (C) hepatic MDA levels in the CON, DEN, DEN + LVD₃, and DEN + HVD₃ group after 8 weeks. DEN significantly increased ALT, AST, and MDA levels. Calcitriol attenuated ALT and AST similarly at both low and high doses, while the high dose produced a greater reduction in MDA than the low dose. Data are presented as mean ± SD, n = 6/group. *p < 0.05 when compared with the CON group, #p < 0.05 when compared with the DEN group, &p < 0.05 when compared with the DEN + LVD3.
Fig 3.
Effects of calcitriol on liver morphology, histological staining, and quantification of Sirius Red-positive area in DEN-induced liver fibrosis.
(A) Representative images of gross liver morphology, (B) H&E staining, (C) Sirius Red staining for collagen deposition, (D) quantification of Sirius Red-positive area (%). Arrowhead indicates fibrotic regions; star indicates collagen deposition. Images were shown at 4x magnification. DEN induced a rough liver surface and increased collagen deposition, while calcitriol improved gross morphology and reduced fibrosis. Data are presented as mean ± SD, n = 6/group. *p < 0.05 when compared with the CON group, #p < 0.05 when compared with the DEN group.
Table 1.
Fibrosis scores across experimental groups.
Fig 4.
Effects of calcitriol on hepatic NF-κB p65 expression in DEN-induced liver fibrosis.
(A) Representative immunohistochemistry images of NF-κB p65 expression. Brown staining indicates positive expression (images shown at 10x magnification). (B) Quantification of strongly positive NF-κB p65 staining (% of total pixels). DEN markedly increased NF-kB p65 expression compared with CON, while both low- and high-dose calcitriol reduced NF-kB p65. Data are presented as mean ± SD, n = 6/group. *p < 0.05 when compared with the CON group, #p < 0.05 when compared with the DEN group.
Fig 5.
Effects of calcitriol on hepatic MMP-12, TIMP-1, α-SMA, and TGF-β1 protein expression in DEN-induced liver fibrosis.
Representative western blot images and quantitative analyses for MMP-12 (A), TIMP-1 (B), α-SMA (C), and TGF-β1 (D). DEN significantly increased MMP-12, α-SMA, and TGF-β1 compared with CON, while calcitriol reduced their expression. TIMP-1 showed no significant changes across groups. Data are presented as mean ± SD, *p < 0.05 when compared with the CON group, #p < 0.05 when compared with the DEN group.
Fig 6.
Effects of calcitriol on hepatic SOD-1, and GPX-1 gene expression in DEN-induced liver fibrosis.
The levels of hepatic SOD-1 (A), and GPX-1 (B) gene expression in all experimental groups. DEN significantly decreased SOD-1 and GPX-1 expression compared with CON. Calcitriol partially enhanced SOD-1 and GPX-1 levels. Data are presented as mean ± SD, n = 6/group. *p < 0.05 when compared with the CON group, #p < 0.05 when compared with the DEN group.