Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

Figure 5 The relationship between calcitriol and nonstructural protein 3-transactivated protein 1. A: RT-qPCR analysis of nonstructural protein 3-transactivated protein 1 (NS3TP1) in mice treated with calcitriol (n = 6); B: Immunohistochemical staining for α-smooth muscle actin (α-SMA) in hepatic tissue, scale bar = 100 μm; C: Image J analysis of immunohistochemical staining for α-SMA, (n = 3); D: Immunofluorescence staining for NS3TP1 in liver tissue, green: NS3TP1 (n = 3), scale bar = 20 μm; E and F: Western blot analysis of NS3TP1 in LX-2 cells stimulated with various concentrations of calcitriol or with 16 μmol/L calcitriol for various durations (n = 3); G: Analysis of luciferase activity between the NS3TP1 promoter and calcitriol in HepG2 cells, calcitriol (L: 8 μmol/L, M: 16 μmol/L, H: 32 μmol/L); H: Western blot analysis of the activity of both above signaling pathways in LX-2 cells treated with different concentrations of calcitriol; I: Collagen I, α-SMA, NS3TP1, and p-p65 in calcitriol-treated LX-2 cells were measured by Western blot and compared to LPS-treated cells; J: Extracellular matrix accumulation was evaluated by Western blot in LX-2 cells stimulated with calcitriol after NS3TP1 overexpression. The data was represented as mean ± SE. ^aP < 0.01 CCl₄ group vs corn oil control group, ^bP < 0.01 calcitriol group vs CCl₄ group; ^cP < 0.0001 pGL4.10-NS3TP1 promoter vs pGL4.10, ^dP < 0.0001 pGL4.10-NS3TP1 promoter + calcitriol (32 μmol/L) vs pGL4.10-NS3TP1 promoter. ECM: Extracellular matrix; LPS: Lipopolysaccharide; NS3TP1: Nonstructural protein 3-transactivated protein 1; TGFβ1: Transforming growth factor 1 beta 1; CCl₄: Carbon tetrachloride.