Alpinetin relieves high-glucose-induced abnormal proliferation of the colon cell line NCM460 by regulating the Nrf2 signaling pathway
Objective To analyze the role of alpinetin in relieving high-glucose-induced abnormal proliferation of the colon cell line NCM460 by regulating the nuclear factor erythroid 2-related factor 2(Nrf2)signaling pathway.Methods The colon cell line NCM460 was induced with blank control,high-level glucose(33 mmol/L),high-level glucose+low-dose alpinetin(25 μmol/L),high-level glucose+medium-dose alpinetin(50 µmol/L),high-level glucose+high-dose alpinetin(100 µmol/L)and high-level glucose+dimethyl fumarate(DMFU,4 µmol/L)for 72 hours.Cell proliferation was detected by CCK-8 assay.Cell proliferation was detected by CCK-8 assay.Reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)levels,relative levels of interleukin-1β(IL-1β),tumor necrosis factor α(TNF-α),IL-8,IL-6 and IL-12 in cell supernatant were measured using the commercial kits.Cell apoptosis and cell cycle distribution were examined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)and propidium iodide(PI)staining,respectively.The mRNA and protein levels of Nrf2 and heme oxygenase-1(HO-1)were detected by real-time quantitative polymerase chain reaction(RTq-PCR)and Western blot,respectively.Results High-glucose-induced NCM460 cells had significantly higher proliferative rate,ROS and MDA levels,relative levels of IL-1β,TNF-α,IL-8,IL-6 and IL-12 in cell supernatant,and proportions of cells arrested in S and G2 phases,but significantly lower SOD and CAT levels,apoptotic rate,proportion of cells arrested in G1 phase and mRNA and protein levels of Nrf2 and HO-1 than those with blank control(P＜0.05).Compared with those induced with high-level glucose,NCM460 cells induced with high-level glucose+alpinetin at varying concentrations or high-level glucose+DMFU had significantly lower proliferative rate,ROS and MDA levels,relative levels of IL-1β,TNF-α,IL-8,IL-6 and IL-12 in cell supernatant,and proportions of cells arrested in S and G2 phases,but higher SOD and CAT levels,apoptotic rate,proportion of cells arrested in G1 phase and mRNA and protein levels of Nrf2 and HO-1 in a dose-dependent manner(P＜0.05).Conclusion Alpinetin significantly inhibits high-glucose-induced inflammation,oxidative stress and proliferation,and promotes apoptosis in NCM460 cells by regulating the Nrf2/HO-1 signaling pathway.
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