Ameliorating liver fibrosis in an animal model using the secretome released from miR-122-transfected adipose-derived stem cells

Figure 2 In vitro experiments validating the effects of miR-122 transfection into Adipose-derived stem cells. A: Western blot analysis showing the expression of fibrotic and antifibrotic markers in miR-122-transfected adipose-derived stem cells (ASCs). miR-122-transfected ASCs showed decreased expression of fibrotic proteins (TGF β1, MMP2, and α-SMA) and increased expression of an antifibrotic protein (TIMP-1) than control ASCs. The graphs below microscopic figures show the relative densities of these markers; B, C: RT-PCR (left) and western blot analysis (right) of LX2 cells for the determination of the thioacetamide (TAA) concentration used for generating in vitro model of liver fibrosis. A TAA concentration of 2.5 mM was used for inducting LX2 cells into fibrosis; D: Effects of MCM in the in vitro model of liver fibrosis. The in vitro model of liver fibrosis was generated by treating human HSCs cells (LX2 cells) with a hepatotoxin (TAA). In western blot analysis (Left), MCM induced the lowest expression of fibrotic markers (MMP2, TGF-β1, and α-SMA) in the TAA-treated LX2 cells. Relative densities of fibrosis-related markers in each group (Right). Values are presented as mean ± standard deviation of three independent experiments. ^aP < 0.05 vs Ct. ^cP < 0.05 between NCM and MCM. α-SMA: Alpha-smooth muscle actin; COL1A1: Collagen type-1 alpha-1; Ct: Control; CM: The secretome obtained from ASCs after 48-h-incubation; MCM: The secretome released from miR-122-transfected ASCs; MMP-1: Metalloproteinases-1; TAA: Thioacetamide; TGF-β: Transforming growth factor-β; TIMP-1: Tissue inhibitor of metalloproteinases-1.