海带中岩藻黄素异构体的分离纯化及其体外抗肿瘤活性研究
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摘要
海带采用乙醇提取色素,通过柱层析分离纯化其中的岩藻黄素,并最终通过高效制备液相色谱制备了三种岩藻黄素的异构体。通过紫外光谱(UV)、核磁共振氢谱(1H-NMR)以及质谱(MS)对岩藻黄素及其异构体进行了结构表征和鉴定。采用乳腺癌细胞MDA-MB-231模型,对岩藻黄素的体外抗肿瘤活性进行了研究。结果表明,岩藻黄素对乳腺癌细胞生长有显著的抑制作用,其中13-顺和13′-顺岩藻黄素组分对癌细胞的抑制活性最强, IC50值达到23.71μg/mL。全反式岩藻黄素和9′-顺岩藻黄素两种异构体IC50值分别为45.30和27.09μg/mL。qPCR试验表明岩藻黄素各异构体可以通过抑制癌细胞NF-κB家族基因表达而抑制乳腺癌细胞生长。
Pigments from Laminaria japonica were extracted using ethyl acetate, and fucoxanthin stereoisomers were purified by column chromatography. Finally, three fucoxanthin stereoisomers were prepared by preparative high performance liquid chromatography. The structures of the fucoxanthin stereoisomers were characterized by ultraviolet(UV) spectroscopy, mass spectrometry(MS), and nuclear magnetic resonance(NMR). The in vitro antitumor activity of the fucoxanthin stereoisomers was analyzed using MDA-MB-231 breast cancer cell line. Results demonstrated that the fucoxanthin stereoisomers possessed significant inhibitory effects against the growth of breast cancer cells. The 13-cis and 13′-cis fucoxanthin mixtures exhibited the highest inhibitory activity, with the IC50 value being 23.71 μg/mL. Moreover, the IC50 values of all trans-fucoxanthin and 9′-cis fucoxanthin mixtures were45.30 and 27.09 μg/mL, respectively. Furthermore, the qPCR results suggested that the fucoxanthin stereoisomers inhibit the growth of breast cancer cells by downregulating the expression of the NF-κB pathway-related genes.
Pigments from Laminaria japonica were extracted using ethyl acetate, and fucoxanthin stereoisomers were purified by column chromatography. Finally, three fucoxanthin stereoisomers were prepared by preparative high performance liquid chromatography. The structures of the fucoxanthin stereoisomers were characterized by ultraviolet(UV) spectroscopy, mass spectrometry(MS), and nuclear magnetic resonance(NMR). The in vitro antitumor activity of the fucoxanthin stereoisomers was analyzed using MDA-MB-231 breast cancer cell line. Results demonstrated that the fucoxanthin stereoisomers possessed significant inhibitory effects against the growth of breast cancer cells. The 13-cis and 13′-cis fucoxanthin mixtures exhibited the highest inhibitory activity, with the IC50 value being 23.71 μg/mL. Moreover, the IC50 values of all trans-fucoxanthin and 9′-cis fucoxanthin mixtures were45.30 and 27.09 μg/mL, respectively. Furthermore, the qPCR results suggested that the fucoxanthin stereoisomers inhibit the growth of breast cancer cells by downregulating the expression of the NF-κB pathway-related genes.
引文
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[3]Satomi Y.Antitumor and cancer-preventative function of fucoxanthin:a marine carotenoid[J].Anticancer Research,2017,37(4):1557-1562.
[4]Masaru Terasaki,Atsushi Hirose,Bhaskar Narayan.Evaluation of recoverable functional lipid components of several brown seaweeds(Phaeophyta)from Japan with special reference to fucoxanthin and fucosterol contents[J]Journal of Phycology,2009,45(4):974-980.
[5]Gerhard Englert,Terje Bj?rnland,Synn?ve LiaaenJensen.1D and 2D NMR-study of some allenic carotenoids of the fucoxanthin series[J].Magnetic Resonance in Chemistry,1990,28(6):519-528.
[6]Wu Hon Yip,Lim Seng Joe,Wan Aida Wan Mustapha,et al.Characterisation and stability of pigments extracted from Sargassum binderi obtained from Semporna,Sabah[J].Sains Malaysiana,2014,43(9):1345-1354.
[7]Sugimura Rumiko,Suad Masatake,Sho Ayumi,et al.Stability of fucoxanthin in dried Undaria pinnatifida(wakame)and baked products(scones)containing wakame powder[J].Food Science and Technology Research,2012,18(5):687-693.
[8]Rwigemera A,Mamelona J,Martin L J.Martin.Comparative effects between fucoxanthinol and its precursor fucoxanthin on viability and apoptosis of breast cancer cell lines MCF-7 and MDA-MB-231[J].Anticancer Research,2015,35(1):207-219.
[9]Nakazawa Y,Sashima T,Hosokawa M,et al.Comparative evaluation of growth inhibitory effect of stereoisomers of fucoxanthin in human cancer cell lines[J].Journal of Functional Foods,2009,1(1):88-97.
[10]Chung T W,Choi H J,Lee J Y,et al.Marine algal fucoxanthin inhibits the metastatic potential of cancer cells[J].Biochemical and Biophysical Research Communications,2013,439(4):580-585.
[11]Wang Jun,Chen Shihui,Xu Shiqiang,et al.In vivo induction of apoptosis by fucoxanthin,a marine carotenoid,associated with down-regulating STAT3/EGFRsignaling in sarcoma 180(S180)xenografts-bearing mice[J].Marine Drugs,2012,10(9):2055-2068.
[12]Satomi Y,Nishino H.Implication of mitogen-activated protein kinase in the induction of G1 cell cycle arrest and gadd45 expression by the carotenoid fucoxanthin in human cancer cells[J].BBA-General Subjects,2009,1790(4):260-266.
[13]Swadesh K Das,Takashi Hashimoto,Kazuki Kanazawa.Growth inhibition of human hepatic carcinoma HepG2cells by fucoxanthin is associated with down-regulation of cyclin D[J].BBA-General Subjects,2008,1780(4):743-749.
[14]Kotake-Nara E,Terasaki,M,Nagao A.Characterization of apoptosis induced by fucoxanthin in human promyelocytic leukemia cells[J].Bioscience Biotechnology and Biochemistry,2005,69(1):224-227.
[15]Martin L.Fucoxanthin and its metabolite fucoxanthinol in cancer prevention and treatment[J].Marine Drugs,2015,13(8):4784-4798.
[16]Zhang Yiping,Fang Hua,Xie Quanling,et al.Comparative evaluation of the radical-scavenging activities of fucoxanthin and its stereoisomers[J].Molecules2014,19(2):2100-2113.
[17]Haugan,J A,Liaaen-Jensen S.Isolation and characterisation of four allenic(6′S)-isomers of fucoxanthin[J].Tetrahedron Letters,1994,35(14):2245-2248.
[18]陈海燕,王志莲,郝敏.SOX9与肿瘤的研究进展[J]中华临床医师杂志(电子版),2016,10(3):424-428.Chen Haiyan,Wang Zhilian,Hao Min.Research advances in SOX9 and its involvement in human malignancies[J].Chinese Journal of Clinicians(Electronic Edition),2016,10(3):424-428.
[19]Hayden M S,Ghosh S.Signaling to NF-kappaB[J]Genes 2004,18(18):2195-2224.
[20]Nakshatri H,Bhat-Nakshatri P,Martin D A,et al.Constitutive activation of NF-kappa B during progression of breast cancer to hormone-independent growth[J]Molecular and Cellular Biology,1997,17(7):3629-3639.