Figure 7. Lysine vasopressin exerts its hepatoprotective effects via V1R-mediated activation of the Wnt/β-catenin /FoxO3a/AKT pathway.

Primary hepatocytes or transfected NCTC cells were subjected to hypoxia (SO₂:3%) for 4 hours and restoration of oxygen supply (95% O₂/5% CO₂) for 4 hours in the presence or absence of lysine vasopressin. A. Western blotting analysis of Wnt3a, β-catenin, Phospho-FoxO3a, FoxO3a, Bim, P27, Phospho-AKT, AKT, BCL-2 and cleaved-caspase-3 in hepatocytes, and densitometric values were shown in Figure S3. B. NCTC cells were transfected with control or V1R lentiviral shRNA and then treated with lysine vasopressin. Western blotting analysis of V1R, Wnt3a, β-catenin, Phospho-FoxO3a, FoxO3a, Bim, P27, Phospho-AKT, AKT, BCL-2 and cleaved-caspase-3 in cells, and densitometric values were shown in Figure S4. C., D. Primary hepatocytes were treated with vehicle or C. SR49059 (10 uM) for 10 minutes or D. XAV939a (1uM) for 24 hours and then subjected to hypoxia (SO₂:3%) for 4 hours and restoration of oxygen supply (95% O₂/5% CO₂) for 4 hours in the presence or absence of lysine vasopressin. Western blot analysis of Wnt3a, β-catenin, Phospho-FoxO3a, FoxO3a, Bim, P27, Phospho-AKT, AKT, BCL-2 and cleaved-caspase-3 in hepatocytes, and densitometric values were shown in Figures S5 and S6, respectively. V1R, arginine vasopressin receptor 1; FoxO3a, forkhead box O 3a; BCL-2, B-cell lymphoma 2.