山姜素调节Nrf2通路对高糖诱导的结肠细胞NCM460异常增殖的影响研究  

Alpinetin relieves high-glucose-induced abnormal proliferation of the colon cell line NCM460 by regulating the Nrf2 signaling pathway

作　　者：江聪 杨保伟 田甜 李夏红 JIANG Cong;YANG Baowei;TIAN Tian(Department of Anorectal,Haikou Hospital of Traditional Chinese Medicine,Hainan,Haikou 570100,China)

机构地区：[1]海南省海口市中医医院肛肠科,570100 [2]海南省海口市中医医院肿瘤科,570100

出　　处：《河北医药》2024年第14期2085-2090,共6页Hebei Medical Journal

基　　金：海南省卫生健康行业科研项目(编号:22A200142)。

摘　　要：目的 研究山姜素对高糖诱导的结肠细胞NCM460异常增殖的影响及对Nrf2通路的调节作用。方法 结肠细胞NCM460分为正常对照组、高糖模型组、山姜素低、中、高剂量组及阳性对照组,高糖模型组加入终浓度为33 mmol/L的葡萄糖,山姜素各剂量组加入终浓度为33 mmol/L的葡萄糖,同时分别加入终浓度为25、50、100μmol/L的山姜素,阳性对照组加入终浓度为33 mmol/L的葡萄糖的同时加入4μmol/L的富马酸二甲酯,培养72 h,CCK-8试剂盒测定NCM460细胞增殖率,使用试剂盒测定NCM460细胞活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)水平及NCM460细胞上清液白介素(IL)-1β、肿瘤坏死因子(TNF)-α、IL-8、IL-6、IL-12水平,原位末端标记法(TUNEL)染色法测定NCM460细胞凋亡率,碘化丙啶(PI)染色法测定NCM460细胞周期分布,实时定量聚合酶链式反应(RTq-PCR)NCM460细胞Nrf2、HO-1 mRNA水平,免疫印迹法(Western blot)测定NCM460细胞Nrf2、HO-1蛋白水平。结果 与正常对照组比较,高糖模型组NCM460细胞增殖率,ROS、MDA水平,NCM460细胞上清液IL-1β、TNF-α、IL-8、IL-6、IL-12水平,NCM460细胞S及G2期比例显著升高(P<0.05),NCM460细胞SOD、CAT水平,NCM460细胞凋亡率及G1期比例,NCM460细胞Nrf2、HO-1 mRNA及蛋白水平显著降低(P<0.05);与高糖模型组比较,山姜素各剂量组及阳性对照组NCM460细胞增殖率,ROS、MDA水平,NCM460细胞上清液IL-1β、TNF-α、IL-8、IL-6、IL-12水平,NCM460细胞S及G2期比例显著降低(P<0.05),NCM460细胞SOD、CAT水平,NCM460细胞凋亡率及G1期比例,NCM460细胞Nrf2、HO-1 mRNA及蛋白水平显著升高(P<0.05),且随着山姜素剂量的增加,各项指标变化更优(P<0.05)。结论 山姜素能够显著抑制高糖诱导的NCM460细胞炎症水平,修复NCM460细胞氧化应激损伤,抑制高糖诱导的NCM460细胞的异常增殖并促进其凋亡,其机制可能与调节Nrf2/HO-1通路有关。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,a

关 键 词：山姜素 Nrf2通路 NCM460细胞 异常增殖 

分 类 号：R285[医药卫生—中药学]