Résumé
Objective To observe the effects of Gandouling on reactive oxygen species (ROS) level, the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA and protein of neural stem cells of the mice cultured in high concentration copper. Methods The model of neural stem cells of the mice was cultured in vitro with high concentration copper. The experimental rats were randomly divided into blank control group, model group, and Gandouling low-, medium-, and high-dose groups. Each medication group was given relevant concentration of Gandouling serum for gavage. The MTT was adopted to test proliferation level on neural stem cells; flow cytometer was used to examine the change of ROS level in cells; qPCR was used to measure the expression of Nrf2 mRNA;Western blot was used to measure the change of the level of protein Nrf2 in cells. Results Compared with the blank control group, the proliferation rate of neural stem cells was significantly decreased, ROS levels were significantly increased, and Nrf2 gene and protein expression was significantly decreased (P＜0.01). Compared with the model group, neural stem cells proliferation rate was significantly increased, ROS levels were significantly reduced, and Nrf2 gene and protein expression was significantly increased (P＜0.05, P＜0.01). Conclusion Gandouling can promote the proliferation of neural stem cells in mice by reducing ROS content in high copper-loaded mice and up-regulating Nrf2 expression.
Résumé
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.