Antioxidation of seaweed polysaccharides on H2O2-induced oxidative damage of human embryonic lung fibroblasts MRC-5
LIU Huaman
LIU Xue
JIA Xinhua
ZHANG Xinyue
ZHANG Wei
Abstract：Objective To investigate the effects of seaweed polysaccharides on H2O2-induced oxidative damage of human embryonic lung fibroblasts MRC-5 and its mechanism.Methods MRC-5 cells cultured in vitro were randomly divided into the blank group, H2O2 group, seaweed polysaccharide group, seaweed polysaccharide+H2O2 group, and H2O2+seaweed polysaccharide group.Cells in the blank group were cultured normally, cells in the H2O2 group were treated with H2O2, cells in the seaweed polysaccharide group were treated with seaweed polysaccharide, cells in the seaweed polysaccharide+H2O2 group were treated with seaweed polysaccharide for 1 h, then followed by H2O2, and cells in the H2O2+seaweed polysaccharide group were treated with H2O2 for 1 h, then followed by H2O2.The concentration of H2O2 in each group was 600 umol/L, and the concentration of seaweed polysaccharide was 0.3125 mg/mL.At 0, 24, 48 and 72 h after treatment, the cell proliferation inhibitory rate in each group was determined by MTT assay.The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS) were measured at 24 h.NF-E2-related factor 2 (Nrf2) protein was detected by immunofluorescence assay.The expression levels of Nrf2, Kelch-like ECH-associated protein 1 (Keap1), NADP (H) quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO-1), and CGLC mRNA were detected by RT-PCR.Results The cell inhibition rates of the H2O2 group, seaweed polysaccharide +H2O2 group, and H2O2 + seaweed polysaccharide group were higher than those of the blank group and seaweed polysaccharide group at 24, 48 and 72 h, and those of the seaweed polysaccharide+H2O2 group and H2O2+seaweed polysaccharide group were lower than that of the H2O2 group (all P<0.05).Compared with the blank group and the seaweed polysaccharide group, the levels of MDA and ROS in the H2O2 group, the seaweed polysaccharide+H2O2 group, and the H2O2+seaweed polysaccharide group increased, and the SOD level decreased.Compared with the H2O2 group, the MDA and ROS levels decreased and the SOD level increased in the seaweed polysaccharide+H2O2 group and the H2O2+seaweed polysaccharide group (all P<0.05).Compared with the blank group and the seaweed polysaccharide group, the expression of Nrf2 mRNA and protein in the H2O2 group, the seaweed polysaccharide+H2O2 group, and the H2O2+seaweed polysaccharide group decreased, and the seaweed polysaccharide+H2O2 group and H2O2+seaweed polysaccharide group were higher than the H2O2 group (all P<0.05).Compared with the blank group and the seaweed polysaccharide group, the relative expression of Keap1 mRNA in the H2O2 group, the seaweed polysaccharide+H2O2 group, and the H2O2+seaweed polysaccharide group increased, and the relative expression of NQO1, CGLC, and HO-1 mRNA decreased.Compared with the H2O2 group, the relative expression of Keap1 mRNA in the polysaccharide+H2O2 group and H2O2+seaweed polysaccharide group decreased, and the relative expression of NQO1, CGLC, and HO-1 mRNA increased (all P<0.05).Conclusion Seaweed polysaccharide could inhibit the oxidative stress injury of MRC-5 induced by H2O2 through up-regulating the expression of Nrf2 and activating Nrf2-Keap1-ARE signaling pathway.
Keywords：pulmonary fibrosishuman embryonic lung fibroblasts MRC-5seaweed polysaccharideH2O2oxidative damagenuclear transcription factor E2-related factor 2
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 17-20 )