Heme Oxygenase-1 Suppresses Wnt Signaling Pathway in Nonalcoholic Steatohepatitis-Related Liver Fibrosis

Figure 4

Induction of HO-1 expression suppressed the Wnt signaling pathway in HSC-T6. (a) Expression levels of HO-1 after Hemin administration at 0, 6, 12, and 24 hours in HSC-T6. β-Actin was used as the loading control. Values are ; vs. 0 h. (b) Then, we administrated HSC-T6 with 0, 20, 40, and 60 μM Hemin for 12 hours. With the increased concentration of Hemin, the HO-1 level was not further increased in HSC-T6 at the concentration of 40 μM. β-Actin was used as the loading control. Values are ; vs. 0 μm. (c) The HSC-T6 were treated with 40 μM Hemin for 12 hours, and the expression of Wnt1 and downstream effectors was determined by Western blot. (d) The nuclear and cytoplasmic fractions were isolated to detect the expression of β-catenin. Lamin A and tubulin were used as the marker for nuclear and cytosolic proteins, respectively. (e) The expression of Wnt5a and NFAT5 were determined by Western blot. (f) The protein levels of oxidative stress markers NQO1 and SOD. (g) The flow cytometry data of Annexin V-FITC/PI to assessing apoptosis of HSC-T6 after the administration of Hemin. (h) Hemin inhibited HSC-T6 cell growth as determined by the CCK-8 assay. (i) The protein levels of fibrosis-related genes were performed by Western blot. β-Actin was used as the loading control. Values are ; , vs. the control group.

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