二苯乙烯苷抗氧化和抗炎作用的机制研究

doi:10.3969/j.issn.1004-616x.2020.02.005

Abstract

Abstract: OBJECTIVE: To explore the induction of antioxidant and anti-inflammation effects of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) in LPS-stimulated RAW264.7 macrophages. METHODS: RAW264.7 cells were pretreated with TSG (30,60,120 μmol/L) for 4 hours,and then exposed to 1 μg/mL LPS for 12 hours. In addition,a blank control and two positive control groups were set up. For the positive controls,cells were treated with dexamethasone (100 nmol/L) or brusatol (50 μmol/L) at 4 h prior to LPS stimulation. Morphological changes were captured under a microscope,and the activated cell rate was analyzed. Concentrations of TNF-α and IL-6 in the culture medium were assessed using specific ELISA kits. Levels of superoxide anions and reactive oxygen species within the cells were detected using flow-cytometry after staining with MitoSOX^TM and DCFH-DA,respectively. Moreover,immunofluorescence staining and western-blot methods were used to investigate protein expression and sub-cellular localization of Nrf2 in the cells. Real-time PCR was used to detect expression of Nrf2-related antioxidant enzymes,including HO-1,SOD₂,SOD₁,CAT and GPX-1. RESULTS: Compared with the control group,LPS activated macrophages exhibited elongated cell morphology with pseudopodium-like protrusions. LPS also strongly promoted the level of pro-inflammatory cytokines,including TNF-α and IL-6. However,pretreatment with TSG inhibited LPS-induced ROS production and inflammatory cytokines secretion. Interestingly,the anti-inflammatory effect of TSG was better than Dex. TSG administration promoted Nrf2 protein expression and its translocated into the nucleus. Moreover,TSG alleviated LPS-induced oxidative stress through regulating Nrf2-related antioxidant enzymes,including HO-1,SOD₂ and CAT. Brusatol,a specific inhibitor of Nrf2,decreased TNF-α and IL-6 production in LPS-activated macrophages. CONCLUSION: TSG attenuated LPS-induced oxidative stress and inflammation in RAW264.7 macrophages through activation of Nrf2 signal pathway. Therefore,TSG may be useful as a drug to prevent or to treat inflammation-associated diseases.

Key words: 2, 3, 5, 4'-tetrahydroxystilbene 2-O-β-D-glucoside, antioxidant, anti-inflammation, Nrf2, RAW264.7 cells

CLC Number:

R114

CHEN Zizhuo, XU Yuhang, ZHAO Jiuzhou, ZHU Jingpu, ZHENG Yipeng, LI Wenli, WU Haozhi, HAI Chunxu, YU Weihua. Antioxidant and anti-inflammatory effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in macrophages[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(2): 105-111,117.

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Antioxidant and anti-inflammatory effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in macrophages

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