岩藻黄素对H_2O_2诱导BNL CL.2细胞氧化损伤保护作用研究
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摘要
建立BNL CL.2细胞氧化损伤模型,探讨岩藻黄素对细胞氧化损伤的保护作用。采用H2O2诱导,以MTT法筛选最佳诱导条件建立BNL CL.2氧化损伤模型。用不同浓度(1~20μmol/L)的岩藻黄素处理模型细胞,MTT法检测细胞活力,LDH试剂盒检测细胞乳酸脱氢酶释放量,GSH试剂盒检测胞内还原型谷胱甘肽含量,荧光酶标仪分析岩藻黄素对胞内活性氧(ROS)的清除能力。1 000μmol/L的H2O2处理BNL CL.2细胞24h成功建立了氧化损伤模型。20μmol/L岩藻黄素可显著提高模型细胞的活力(P <0.05);1~20μmol/L岩藻黄素可明显降低模型细胞乳酸脱氢酶释放量(P <0.05),可显著提高细胞内源性抗氧化剂谷胱甘肽的含量(P <0.05);1~10μmol/L岩藻黄素可显著降低模型细胞内ROS的含量(P <0.05)。岩藻黄素具有抗氧化作用,可抑制H2O2引起的BNL CL.2细胞的氧化损伤。
The aim of this study was to investigate the protective effect of fucoxanthin against H2 O2-induced oxidative damage through establishing oxidative damage model of BNL CL.2 cells.BNL CL.2 cells were induced by H2 O2 and MTT method was used to screen the appropriate concentration of H2 O2 to establish oxidative damage model.BNL CL.2 cells were cultured in the medium supplemented with different concentration of fucoxanthin(1~20μmol/L).The effect of fucoxanthin on the cell viability was detected by MTT test.LDH release assay kit was used to detect the lactate dehydrogenase release of the cells.GSH assay kit was used to detect the GSH content of the cells.The reactive oxygen species(ROS)scavenging activity of FUCO was tested by fluorescence microplate reader.The model of oxidative damage in BNL CL.2 cells was successfully established by 1 000μmol/L H2 O2 treatment for 24 h.Fucoxanthin at the concentration of 20μmol/L significantly increased cell viability of the oxidatively damaged cells(P<0.05).LDH Release of the H2 O2-treated cells was significantly decreased(P<0.05),but the glutathione content was significantly increased by 1~20μmol/L fucoxanthin.The level of intracellular ROS was significantly decreased by 1~10μmol/L fucoxanthin(P<0.05).Fucoxanthin has antioxidant activity and can protect the BNL CL.2 cells from H2 O2-induced oxidative damage.
The aim of this study was to investigate the protective effect of fucoxanthin against H2 O2-induced oxidative damage through establishing oxidative damage model of BNL CL.2 cells.BNL CL.2 cells were induced by H2 O2 and MTT method was used to screen the appropriate concentration of H2 O2 to establish oxidative damage model.BNL CL.2 cells were cultured in the medium supplemented with different concentration of fucoxanthin(1~20μmol/L).The effect of fucoxanthin on the cell viability was detected by MTT test.LDH release assay kit was used to detect the lactate dehydrogenase release of the cells.GSH assay kit was used to detect the GSH content of the cells.The reactive oxygen species(ROS)scavenging activity of FUCO was tested by fluorescence microplate reader.The model of oxidative damage in BNL CL.2 cells was successfully established by 1 000μmol/L H2 O2 treatment for 24 h.Fucoxanthin at the concentration of 20μmol/L significantly increased cell viability of the oxidatively damaged cells(P<0.05).LDH Release of the H2 O2-treated cells was significantly decreased(P<0.05),but the glutathione content was significantly increased by 1~20μmol/L fucoxanthin.The level of intracellular ROS was significantly decreased by 1~10μmol/L fucoxanthin(P<0.05).Fucoxanthin has antioxidant activity and can protect the BNL CL.2 cells from H2 O2-induced oxidative damage.
引文
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[4] Lin H V,Tsou Y C,Chen Y T,et al.Effects of low-molecularweight fucoidan and high stability fucoxanthin on glucose homeostasis,lipid metabolism,and liver function in a mouse model of typeⅡdiabetes[J].Marine Drugs,2017,15(4):113-127.
[5] Maeda H.Nutraceutical effects of fucoxanthin for obesity and diabetes therapy:A review[J].Journal of Oleo Science,2015,64(2):125-132.
[6] Raposo M F D J,Rui M S C D M.Carotenoids from marine microalgae:A valuable natural source for the prevention of chronic diseases[J].Marine Drugs,2015,13(8):5128-5155.
[7] Sachindra N M,Sato E,Maeda H,et al.Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites[J].Journal of Agricultural&Food Chemistry,2007,55(21):8516-8522.
[8] Heo S J,Ko S C,Kang S M,et al.Cytoprotective effect of fucoxanthin isolated from brown algae Sargassum siliquastrum,against H2O2-induced cell damage[J].European Food Research and Technology,2008,228(1):145-151.
[9] Liu Chengling,Chiu Yuting,Hu Miaolin.Fucoxanthin enhances HO-1and NQO1expression in murine hepatic BNL CL.2cells through activation of the Nrf2/ARE system partially by its pro-oxidant activity[J].Journal of Agricultural and Food Chemistry,2011,59:11344-11351.
[10]刘甜甜,赵海峰.H2O2诱导大鼠原代海马神经细胞建立氧化损伤细胞模型[J].卫生研究,2014,43(5):719-723.Liu Tiantian,Zhao Haifeng.H2O2-induced oxidative damage in primary cultured rat hippocampal neurons[J].Health Research,2014,43(5):719-723.
[11] Ha A W,Na S J,Kim W K.Antioxidant effects of fucoxanthin rich powder in rats fed with high fat diet[J].Nutr Res Pract,2013,7(6):475-480.
[12]兰爱平,梅卫义,孟金兰,等.硫化氢通过抑制p38MAPK保护PC12细胞对抗化学性缺氧损伤[J].中国药理学通报,2010,26(10):1339-1344.Lan Aiping,Mei Weiyi,Meng Jinlan,et al.Hydrogen sulfide protects PC12cells against chemical hypoxic injury by inhibiting p38MAPK[J].Chinese Pharmacological Bulletin,2010,26(10):1339-1344.
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