Effects of high copper on Wnt/β-catenin signaling pathway in hepatocytes / 中国神经精神疾病杂志

ABSTRACT

Objective To study the effect of high copper on Wnt /β-catenin signaling pathway and oxidative stress. Methods BRL-3A cells were incubated with different concentrations of CuSO4.The cell growth and proliferation wre assessed using MRR method.The production of reactive oxygen species(ROS)and mitochondrial membrane potential (JC-1) were examined usingf flow cytometry. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were detected by microplate reader. The expression of protein was detected by Western Blot. Results①The results of MTT showed that CuSO4inhibited the growth and proliferation of BRL-3A cells in a time- and concentration-dependent manner(P<0.01).②Flow cytometry results showed that CuSO4induced a large amount of ROS and significantly decreased the fluorescence intensity of JC-1 in BRL-3A cells (P<0.01). ③ Microplate reader results showed that CuSO4increased the content of MDA and decreased the activity of SOD (P<0.05).④Western blotting assay showed that CuSO4significantly decreased the total expression levels of β-catenin and p-Ser 9-GSK-3β protein as well as nuclear levels of p-(S33+S37)-β-catenin and c-Myc (P<0.01) and increased expression levels of GSK-3β、DKK1、Dishevelled3 protein in BRL-3A cells. Conclusion High copper can induce oxidative stress and induce Wnt /β-catenin signaling pathway to deactivate liver cells,leading to hepatocellular injury.