Gallic Acid Activates Nrf-2 Signaling Pathway to Alleviate High Glucose-Induced Oxidative Stress and Inflammation of NIH-3T3 Cells
Objective:To investigate the role of gallic acid(GA)in activating the nuclear factor erythroid 2-related factor 2(Nrf-2)pathway to re-duce inflammation and oxidative stress induced by high glucose in NIH-3T3 cells and restore cell function.Methods:The proliferation activity of NIH-3T3 cells exposed to different concentrations of glucose and treated with GA for 24 h was examined by the Cell-Counting Kit-8(CCK-8)method,and thus the optimal glucose concentration for modeling and GA concentration for intervention were determined.The proliferation viability of NIH-3T3 cells exposed to high glucose and treated with GA was detected by the CCK-8 method.The DCFH-DA fluorescent probe was used to measure the content of reactive oxygen species(ROS)in NIH-3T3 cells.Immunofluorescence was used to detect the nuclear translocation of Nrf-2.Western blotting and RT-PCR were employed to determine the protein and mRNA levels,respectively,of the proteins in the Nrf-2 and nuclear factor-kappa B(NF-κB)signaling pathways.In addition,the expression of collagen type Ⅰ(COL-Ⅰ)was measured by Western blotting.Results:The cells were modeled with 18 mg/mL glucose and treated with 0.85 μg/mL GA.Compared with the normal con-trol group,the model group showed decreased cell proliferation(P＜0.01),down-regulated protein levels of Nrf-2,KEAP-1,HO-1 and IκB(P＜0.01),up-regulated protein levels of P65 and p-P65(P＜0.05 or P＜0.01),increased the ratio of p-P65/P65(P＜0.01),up-regulated mRNA levels of Keap1,Nrf2,Nqo1,P65,Ii6 and Tnfα(P＜0.05),down-regulated mRNA levels of Ho1,Sod and Iκb(P＜0.05 or P＜0.01),increased ROS content,decreased Nrf-2 expression and nuclear translocation(P＜0.05 or P＜0.01),and attenuated COL-Ⅰ synthesis(P＜0.01).Compared with the model group,GA intervention restored cell proliferation(P＜0.01),up-regulated the proteins expression of Nrf-2,HO-1 and IκB(P＜0.05 or P＜0.01),down-regulated protein level of p-P65 and ratio of p-P65/P65(P＜0.01),up-regulated mRNA levels of Nrf2,Ho1,Sod,Nqo1,Cat,Gst and Iκb(P＜0.05 or P＜0.01),down-regulated mRNA levels of Keap1,P65,Ii6 and Tnfα(P＜0.05 or P＜0.01),reduced ROS content,increased the Nrf-2 expression and nuclear translocation(P＜0.05 or P＜0.01),and enhanced COL-Ⅰ synthesis(P＜0.01).Conclusion:GA can scavenge ROS and reduce oxidative stress and inflammation in the NIH-3T3 cell by activating Nrf-2 signa-ling pathway,thus repairing the NIH-3T3 cell damage caused by high glucose.
Gallic acidDiabetic woundInjury induced by high glucoseoxidative stressInflammation
NETL
NSTL
万方数据
