岩藻黄素抗H_2O_2诱导WI-38细胞早衰的作用研究
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摘要
目的探讨岩藻黄素是否具有抑制H_2O_2引起的WI-38细胞早衰的活性。方法 WI-38细胞在添加岩藻黄素的培养基中培养一定时间后经H_2O_2处理诱导发生早衰,用MTT法检测细胞活力,SA-β-半乳糖苷酶染色法检测细胞内衰老相关的SA-β-半乳糖苷酶活性。结果 300μmol·L-1 H_2O_2处理WI-38细胞20min可成功建立早衰细胞模型。与空白对照组相比,H_2O_2处理组细胞活力明显降低,SA-β-半乳糖苷酶阳性细胞数明显增多。5和10μmol·L-1岩藻黄素组细胞活力明显高于H_2O_2处理组,SA-β-半乳糖苷酶阳性细胞数较H_2O_2组明显减少。结论岩藻黄素能够抑制由H_2O_2处理引起的细胞早衰,具有一定的抗衰老作用。
Objective To investigate the anti-aging effect of fucoxanthin on H_2O_2-induced premature senescence in WI-38 human diploid fibroblasts.Methods The cell viability tested by MTT method and the morphological observation under microscope were used to screen the conditions for the establishment of WI-38 cells model with H_2O_2-induced premature senescence.WI-38 cells were cultured in medium supplemented with different concentrations of fucoxanthin and then treated with H_2O_2 to induce premature senescence.The cell viability and senescence-associated SA-β-galactosidase activity assessed by SA-β-galactosidase staining were tested to evaluate cellular senescence in different groups.Results The premature senescence of WI-38 cells could be induced by treatment with 300μmol·L~(-1) H_2O_2 for 20 min.Compared with control group,the cell viability of H_2O_2-treated group was significantly decreased,and the number of SA-β-galactosidase positive cells was significantly increased.However,in the 5 and 10μmol·L~(-1) FUCO groups,the cell viability was significantly higher than that of the H_2O_2-treated group,the numbers of SA-β-galactosidase positive cells was significantly decreased than that of the H_2O_2-treated group.Conclusion FUCO could protect WI-38 cells from H_2O_2-induced premature senescence,it had anti-aging effect.
Objective To investigate the anti-aging effect of fucoxanthin on H_2O_2-induced premature senescence in WI-38 human diploid fibroblasts.Methods The cell viability tested by MTT method and the morphological observation under microscope were used to screen the conditions for the establishment of WI-38 cells model with H_2O_2-induced premature senescence.WI-38 cells were cultured in medium supplemented with different concentrations of fucoxanthin and then treated with H_2O_2 to induce premature senescence.The cell viability and senescence-associated SA-β-galactosidase activity assessed by SA-β-galactosidase staining were tested to evaluate cellular senescence in different groups.Results The premature senescence of WI-38 cells could be induced by treatment with 300μmol·L~(-1) H_2O_2 for 20 min.Compared with control group,the cell viability of H_2O_2-treated group was significantly decreased,and the number of SA-β-galactosidase positive cells was significantly increased.However,in the 5 and 10μmol·L~(-1) FUCO groups,the cell viability was significantly higher than that of the H_2O_2-treated group,the numbers of SA-β-galactosidase positive cells was significantly decreased than that of the H_2O_2-treated group.Conclusion FUCO could protect WI-38 cells from H_2O_2-induced premature senescence,it had anti-aging effect.
引文
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[19]Piechota M,Sunderland P,Wysocka A,et al.Is senescence-associatedβ-galactosidase a marker of neuronal senescence[J].Oncotarget,2016,7(49):81099-81109.
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[2]Zheng L,Liu M,Zhai G Y,et al.Antioxidant and anti-ageing activities of mycelia zinc polysaccharide from Pholiota namekv SW-03[J].J Sci Food Agric,2015,95(15):3117-3126.
[3]夏世金,孙涛,吴俊珍.自由基,炎症与衰老[J].实用老年医学,2014,28(2):100-103.
[4]Salminen A,Kaarniranta K,Kauppinen A.Crosstalk between oxidative stress and Sirt1:Impact on the aging process[J].Int J Mol Sci,2013,14(2):3834-3859.
[5]Takahashi K,Hosokawa M,Kasajima H,et al.Anticancer effects of fucoxanthin and fucoxanthinol on colorectal cancer cell lines and colorectal cancer tissues[J].Oncol Lett,2015,10(3):1463-1467.
[6]Maoka T,Nishino A,Yasui H,et al.Anti-oxidative Activity of Mytiloxanthin,a Metabolite of Fucoxanthin in Shellfish and Tunicates[J].Mar Drugs,2016,14(5):93-94.
[7]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.
[8]Lin H V,Tsou Y C,Chen Y T,et al.Effects of Low-Molecular-Weight Fucoidan and High Stability Fucoxanthin on Glucose Homeostasis,Lipid Metabolism,and Liver Function in a Mouse Model of TypeⅡDiabetes[J].Mar Drugs,2017,15(4):113-126.
[9]Maeda H.Nutraceutical effects of fucoxanthin for obesity and diabetes therapy:a review[J].J Oleo Sci,2015,64(2):125-132.
[10]Lashmanova E,Proshkina E,Zhikvivetskaya S,et al.Fucoxanthin increase lifespan of Drosophila melanogaster and Caenorhabditis elegans[J].Pharmacol Res,2015,100:228-241.
[11]Lim Y,Lee E,Lee J,et al.Down-regulation of Asymmetric Arginine Methylation During Replicative and H2O2-induced Premature Senescence in WI-38 Human Diploid Fibroblasts[J].J Biochem,2008,144(4):523-529.
[12]徐薇,庄志雄,杨建平,等.过氧化氢诱导HPF细胞早衰模型与细胞复制性衰老的生物学性状比较研究[J].毒理学杂志,2013,27(3):164-168.
[13]郝莉,徐玉英,郭春霞,等.银杏酮酯抗H2O2诱导衰老海马神经元氧化DNA损伤的作用研究[J].中药药理与临床,2015,31(3):83-88.
[14]蒲秀瑛,于双,樊文博,等.归芪多糖对早衰WI-38细胞端粒酶活性的影响[J].中国中医药信息杂志,2015,22(11):50-53.
[15]Kim B,Park K Y,Kim H Y,et al.Anti-aging effects and mechanisms of kimchi during fermentation under stress-induced premature senescence cellular system[J].Food Sci Biotechnol,2011,20(3):643-649.
[16]Pu X,Yu S,Fan W,et al.Guiqi polysaccharide protects the normal human fetal lung fbroblast WI-38cells from H2O2-induced premature senescence[J].Int J Clin Exp Patho,2015,8(5):4398-4407.
[17]谢健,童培建,单乐天.H2O2氧化应激诱导兔椎间盘髓核细胞衰老的研究[J].中国骨伤,2013,26(4):332-335.
[18]Wang Y,Meng A M,Zhou D H.Inhibition of phosphatidylinostol 3-kinase uncouples H2O2-induced senescent phenotype and cell cycle arrest in normal human diploid fibroblasts[J].Exp Cell Res,2004,298(1):188-196.
[19]Piechota M,Sunderland P,Wysocka A,et al.Is senescence-associatedβ-galactosidase a marker of neuronal senescence[J].Oncotarget,2016,7(49):81099-81109.
[20]Xiao X,Chen Z K,Ding Y Q,et al.Senescence-associated beta galactosidase expression in rat schwann cells after chronic denervation[J].四川大学学报:医学版,2016,47(3):351-355.