To establish whether vitamin C deficiency induces up-regulation of PPAR-γ expression in the liver of SMP30 KO mice, we performed an additional animal experiment using 8-week-old WT mice and SMP30 KO mice as follows: a WT group (n = 6), a SMP30 KO group without vitamin C (n = 6), and a vitamin C-treated SMP30 KO group (n = 6) for a period of 16 Hydroxychloroquine datasheet weeks. Vitamin C was

provided in the drinking water (L-ascorbic acid, 1.5 g/L) during the experimental period. Following immunoblot analysis, as expected, vitamin C-treated SMP30 KO mice revealed significantly decreased PPAR-γ expression levels in the liver tissue compared with nonvitamin C-treated SMP30 KO mice (Fig. 6A,B). These results indicate that vitamin C might be involved directly in the regulation of PPAR-γ expression in the liver. Therefore, it is believed that higher expression levels of PPAR-γ were caused by vitamin

C deficiency in SMP30 KO mice. To assess reproducibility and whether vitamin C supplement restores CCl4-induced liver fibrosis in SMP30 KO mice, we performed another set of animal experiments using 8-week-old, WT mice, and SMP30 KO mice as follows; WT group (n = 7), CCl4-treated WT group (n = 7), CCl4+vitamin C WT group (n = 7), SMP30 KO group (n = 5), CCl4-treated SMP30 KO group (n = 5), and learn more CCl4+vitamin C SMP30 KO group (n = 5), for an experiment period of 16 weeks. Interestingly, significantly increased liver fibrosis, measured by morphometry based on Masson’s trichrome stain, was observed in the CCl4 + vitamin C SMP30 KO group compared with the CCl4-treated SMP30 KO group, whereas the WT mice showed no noticeable differences between the CCl4-treated WT group and the CCl4 + vitamin C WT group (Fig. 7A,B). These histological findings were further confirmed by measurement of the hydroxyproline content (Fig. 7C) and α-SMA expression level (Fig. 7D,E) in the liver, which demonstrated that vitamin C supplements restore CCl4-induced liver fibrosis in SMP30 KO mice. Taken together, these data

suggest that vitamin C deficiency suppresses HSC activation following a CCl4-induced click here liver injury. In this study we demonstrate for the first time that up-regulation of PPAR-γ expression by way of vitamin C deficiency inhibits HSC activation in SMP30 KO mice. We were led to accept that vitamin C deficiency caused by the absence of SMP30 can lead to: (1) ameliorated liver fibrosis; (2) inhibition of nuclear translocation of p-Smad2/3 in HSCs and hepatocytes; (3) higher PPAR-γ expression levels in SMP30 KO HSCs; (4) up-regulation of PPAR-γ, which is associated with vitamin C deficiency. Moreover, we confirmed that vitamin C supplement restores liver fibrosis in vitamin C-deficient SMP30 KO mice.