山东医药2017，Vol.57Issue(32)：17-20,4.DOI:10.3969/j.issn.1002-266X.2017.32.005
海藻多糖对H2O2诱导人胚肺成纤维细胞MRC-5氧化损伤的影响及其机制
Antioxidation of seaweed polysaccharides on H2O2-induced oxidative damage of human embryonic lung fibroblasts MRC-5
摘要
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.关键词
肺纤维化/人胚肺成纤维细胞MRC-5/海藻多糖/过氧化氢/氧化损伤/核转录因子E2相关因子2Key words
pulmonary fibrosis/human embryonic lung fibroblasts MRC-5/seaweed polysaccharide/H2O2/oxidative damage/nuclear transcription factor E2-related factor 2分类
医药卫生引用本文复制引用
刘骅漫,刘学,贾新华,张心月,张伟..海藻多糖对H2O2诱导人胚肺成纤维细胞MRC-5氧化损伤的影响及其机制[J].山东医药,2017,57(32):17-20,4.基金项目
国家自然科学基金资助项目(81273704) （81273704）
"泰山学者"建设工程(ts20110819). （ts20110819）
