Adeno-associated virus mediated interferon-gamma inhibits the progression of hepatic fibrosis in vitro and in vivo

Figure 1 Structure of recombinant plasmid AAV-INF-γ. Human INF-γ cDNA obtained from RT-PCR was cloned into AAV plasmid, which contains a CAG promoter, WPRE enhancer and BGH polyA.

Figure 2 The expression of INF-γ in rAAV-INF-γ infected HSCs. A: RT-PCR analysis of INF-γ expression in rAAV-INF-γ infected HSCs. The untreated and rAAV-eGFP infected HSCs did not show any PCR band; B: Time course of the INF-γ expression in the HSC supernatants after rAAV-INF-γ infection. (Data are mean±SD, n = 6).

Figure 3 The expression of α-SMA in different group of primary cultured HSCs. Expressions of α-SMA were determined by immunocytochemical method. A: Immunocytochemistry of untreated HSCs without primary antibody as the negative control; B: Immunocytochemistry of untreated HSCs; C: Immunocytochemistry of rAAV-eGFP infected HSCs; D: Immunocytochemistry of rAAV-INF-γ infected HSCs.

Figure 4 The mRNA expression of fibrosis related cytokines and proteases in different group of primary cultured HSCs. A: mRNA expression of TGF-β, MMP-13 and TIMP-1 in HSCs were examined by RT-PCR method; B: Densitometric units were calculated by dividing the density of individual bands by that of GAPDH. Closed column, untreated. Lined column, rAAV-eGFP induced. Dotted column, rAAV-INF-γ induced. (Data are mean±SD, n = 8, ^aP < 0.05).

Figure 5 Improvement of CCl₄ induced experimental hepatic fibrosis by rAAV-INF-γ treatment in histology analysis. Rat liver section in each group stained with Hematoxylin Eosin: A: No treatment; B: CCl₄ (3 wk); C: CCl₄/saline; D: CCl₄/rAAV-eGFP; E: CCl₄/rAAV-INF-γ. (Magnification 200 ×).

Figure 6 The mRNA expression of fibrosis related cytokines and proteases in different group of rat liver. A: mRNA expressions of TGF-β, MMP-13 and TIMP-1 in the rat liver were examined by RT-PCR method; B: Densitometric units were calculated by dividing the density of individual bands by that of GAPDH. Closed column, untreated. Lined column, rAAV-eGFP induced. Dotted column, rAAV-INF-γ induced. (Data are mean±SD, n = 8, ^aP < 0.05).