山楂原花青素对人肝癌SMMC-7721细胞增殖和凋亡的影响
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摘要
目的:探讨山楂原花青素提取物、表儿茶素和原花青素B2对人肝癌SMMC-7721细胞增殖及凋亡的影响。方法:体外培养SMMC-7721细胞,噻唑蓝法检测不同质量浓度山楂原花青素对细胞增殖的影响;4’,6-二脒基-2-苯基吲哚染色法观察细胞凋亡,Western blot法检测细胞凋亡相关蛋白的表达;检测每组细胞中过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、Na~+K~+-ATP酶活力和丙二醛(malondialdehyde,MDA)含量。结果:山楂原花青素提取物、表儿茶素和原花青素B_2呈时间和质量浓度依赖性地显著抑制SMMC-7721细胞的增殖(P<0.05),促进细胞凋亡;与对照组相比,SMMC-7721细胞中SOD、CAT、GSH-Px、Na~+K~+-ATP酶活力降低,MDA含量升高;细胞凋亡相关蛋白半胱天冬酶-8、半胱天冬酶-3、Bcl-2相关X蛋白表达量和多聚二磷酸腺苷核糖聚合酶片段化增加,Bcl-2表达量降低。结论:山楂原花青素可通过影响抗氧化酶活力和细胞凋亡相关蛋白来抑制SMMC-7721细胞增殖,促进细胞凋亡。
Objective: To investigate the effect of hawthorn proanthocyanidins extract, epicatechin and proanthocyanidin B2 on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells. Methods: SMMC-7721 cells were cultured in vitro. The effect of different concentrations of hawthorn proanthocyanidins on cell proliferation and apoptosis was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay and 4',6-diamidino-2-phenylindole(DAPI) staining, respectively. The expression of apoptosis-related proteins was detected by Western blotting. The activities of catalase(CAT), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and Na~+K~+-ATPase and malondialdehyde(MDA) level in cells from each group were determined. Results: Hawthorn proanthocyanidins extract, epicatechin and proanthocyanidin B_2 all significantly inhibited the proliferation of SMMC-7721 cells in a time-dependent and concentrationdependent manner(P < 0.05) and induced cell apoptosis. Compared with the control group, hawthorn proanthocyanidins extract, epicatechin and proanthocyanidin B_2 decreased the activity of SOD, CAT, GSH-Px and Na~+K~+-ATPase and increased the level of MDA in SMMC-7721 cells. The expression of apoptosis-related proteins casepase-8, caspase-3,Bcl-2-associated X protein and the fragment of poly(ADP-ribose) polymerase were increased, while the expression of Bcl-2 was decreased. Conclusion: Hawthorn proanthocyanidins can inhibit the proliferation of SMMC-7721 cells and induce apoptosis through regulating antioxidant activity and apoptosis-related proteins.
Objective: To investigate the effect of hawthorn proanthocyanidins extract, epicatechin and proanthocyanidin B2 on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells. Methods: SMMC-7721 cells were cultured in vitro. The effect of different concentrations of hawthorn proanthocyanidins on cell proliferation and apoptosis was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay and 4',6-diamidino-2-phenylindole(DAPI) staining, respectively. The expression of apoptosis-related proteins was detected by Western blotting. The activities of catalase(CAT), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and Na~+K~+-ATPase and malondialdehyde(MDA) level in cells from each group were determined. Results: Hawthorn proanthocyanidins extract, epicatechin and proanthocyanidin B_2 all significantly inhibited the proliferation of SMMC-7721 cells in a time-dependent and concentrationdependent manner(P < 0.05) and induced cell apoptosis. Compared with the control group, hawthorn proanthocyanidins extract, epicatechin and proanthocyanidin B_2 decreased the activity of SOD, CAT, GSH-Px and Na~+K~+-ATPase and increased the level of MDA in SMMC-7721 cells. The expression of apoptosis-related proteins casepase-8, caspase-3,Bcl-2-associated X protein and the fragment of poly(ADP-ribose) polymerase were increased, while the expression of Bcl-2 was decreased. Conclusion: Hawthorn proanthocyanidins can inhibit the proliferation of SMMC-7721 cells and induce apoptosis through regulating antioxidant activity and apoptosis-related proteins.
引文
[1] NACZK M, SHAHIDI F. Extraction and analysis of phenolics in food[J]. Journal of Chromatography A, 2004, 1054(1/2): 95-111. DOI:10.1016/j.chroma.2004.08.059.
[2] YANG Y, CHIEN M. Characterization of grape procyanidins using high-performance liquid chromatography/mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry[J]. Journal of Agricultural and Food Chemistry, 2000, 48(9): 3990-3996. DOI:10.1021/jf000316q.
[3] CUI T, LI J Z, KAYAHARA H, et al. Quantification of the polyphenols and triterpene acids in Chinese hawthorn fruit by high-performance liquid chromatography[J]. Journal of Agricultural and Food Chemistry, 2006, 54(13): 4574-4581. DOI:10.1021/jf060310m.
[4] PUIGGRòS F, SALVADò M J, BLADé C, et al. Differential modulation of apoptotic processes by proanthocyanidins as a dietary strategy for delaying chronic pathologies[J]. Critical Reviews in Food Science and Nutrition, 2014, 54(3): 277-291. DOI:10.1080/10408398.2011.565456.
[5]禹华娟,孙智达,谢笔钧.莲原花青素在油脂体系中的抗氧化作用[J].中国农业科学, 2010, 43(10): 2132-2140. DOI:10.3864/j.issn.0578-1752.2010.10.020.
[6]谢文利,晋玉章,万宗明,等.聚果多酚抗衰老作用的实验研究[J].食品科学, 2009, 30(9): 207-209.
[7]PRASANNA R,HARISH C C,PICHAI R,et al.Anti-cancer effect of Cassia auriculata leaf extract in vitro through cell cycle arrest and induction of apoptosis in human breast and larynx cancer cell lines[J]. Cell Biology International, 2009, 33(2): 127-134. DOI:10.1016/j.cellbi.2008.10.006.
[8] GAO J, INAGAKI Y, LIU Y. Research progresses on flavonoids isolated from traditional Chinese medicine in treatment of Alzheimer’s disease[J]. Intractable and Rare Diseases Research, 2013, 2(1): 3-10. DOI:10.5582/irdr.2013.v2.1.3.
[9] WANG L, FUMOTO T, MASUMOTO S, et al. Regression of atherosclerosis with apple procyanidins by activating the ATP-binding cassette subfamily a member 1 in a rabbit model[J]. Atherosclerosis, 2017,258:56-64.DOI:10.1016/j.atherosclerosis.2017.01.032.
[10] LIANG Y, WANG J, GAO H Q, et al. Beneficial effects of grape seed proanthocyanidin extract on arterial remodeling in spontaneously hypertensive rats via protecting against oxidative stress[J]. Molecular Medicine Reports, 2016, 14(4): 3711-3718. DOI:10.3892/mmr.2016.5699.
[11] LIU Z Q, YANG Q L, ZHANG C S, et al. Study on antioxidant activity of proanthocyanidins from peanut skin[J]. Advanced Materials Research,2011,197/198:1582-1586.DOI:10.4028/www.scientific.net/AMR.197-198.1582.
[12]JORD?O A M,GON?A LVES F J,CORREIAA C,et al.Proanthocyanidin content, antioxidant capacity and scavenger activity of Portuguese sparkling wines (Bairrada appellation of origin)[J]. Journal of the Science of Food and Agriculture, 2010, 90(12): 2144-2152.DOI:10.1002/jsfa.4064.
[13] WANG Xinghui, HUANG Lingling, YU Tingting, et al. Effects of oligomeric grape seed proanthocyanidins on heart, aorta, kidney in DOCA-salt mice: role of oxidative stress[J]. Phytotherapy Research, 2013, 27(6): 869-876. DOI:10.1002/ptr.4793.
[14] AN J, CHEN C, WANG X, et al. Oligomeric proanthocyanidins alleviate hexabromocyclododecane nduced cytotoxicity in HepG2 cells through regulation on ROS formation and mitochondrial pathway[J]. Toxicology in Vitro, 2014, 28(2): 319-326. DOI:10.1016/j.tiv.2013.11.009.
[15] FARIA A, CALHAU C, DE FREITAS V, et al. Procyanidins as antioxidants and tumor cell growth modulators[J]. Journal of Agricultural and Food Chemistry, 2006, 54(6): 2392-2397. DOI:10.1021/jf0526487.
[16] CHEN J Y, XU Z, ZHU W, et al. Novel proanthocyanidin dimer analogues with the C-ring-opened diaryl-propan-2-gallate structural unit and enhanced antioxidant activities[J].Journal of Functional Foods,2016,21:290-300.DOI:10.1016/j.jff.2015.12.023.
[17] AGUILERA Y, ESTRELLA I, BENITEZ V, et al. Bioactive phenolic compounds and functional properties of dehydrated bean flours[J]. Food Research International, 2011, 44(3): 774-780. DOI:10.1016/j.foodres.2011.01.004.
[18]赵丹,曾新安,徐中岳.不同聚合度葡萄皮原花青素的抗氧化活性[J].食品与发酵工业, 2012, 38(4): 4104-4108.DOI:10.13995/j.cnki.11-1802/ts.2012.04.009.
[19] LIZARRAGA D, LOZANO C, BRIEDé J J, et al. The importance of polymerization and galloylation for the antiproliferative properties of procyanidin-rich natural extracts[J]. The FEBS Journal, 2007, 274(18): 4802-4811.DOI:10.1111/j.1742-4658.2007.06010.x.
[20] LOZANO C, JULIá L, JIMéNEZ A, et al. Electron-transfer capacity of catechin derivatives and influence on the cell cycle and apoptosis in HT29 cells[J]. The FEBS Journal, 2006, 273(11): 2475-2486. DOI:10.1111/j.1742-4658.2006.05255.x.
[21] BRAY F, FERLAY J, SOERJOMATARAM L, et al. Global Cancer Statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA: A Cancer Journal for Clinicians, 2018, 68(6): 394-424. DOI:10.3322/caac.21492.
[22] CUI T, NAKAMURA K, TIAN S, et al. Polyphenolic content and physiological activities of Chinese hawthorn extracts[J]. Bioscience, Biotechnology, and Biochemistry, 2006, 70(12): 2948-2956. DOI:10.1271/bbb.60361.
[23]谭卫东,金红,罗弟祥,等.抗肿瘤药物筛选中MTT法和SRB法的比较[J].天然产物研究与开发, 1999, 11(3): 17-22. DOI:10.16333/j.1001-6880.1999.03.004.
[24]董成龙,游娜,许向红,等. 白藜芦醇对人甲状腺乳头状癌IHH4细胞增殖和凋亡的影响[J].南京医科大学学报(自然科学版), 2015, 35(8): 1096-1103. DOI:10.7655/NYDXBNS20150809.
[25] ROY A M, BALIGA M S, ELMETS C A, et al. Grape seed proanthocyanidins induce apoptosis through p53, Bax, and Caspase 3 pathways[J]. NeoPlasia, 2005, 7(1): 24-36.DOI:10.1593/neo.04412.
[26]SHAHIDI F,AMBIGAIPALAN P.Phenolics and polyphenolics in foods, beverages and spices: antioxidant activity and health effects: a review[J]. Journal of Functional Foods, 2015, 18: 820-897. DOI:10.1016/j.jff.2015.06.018.
[27]WANG S N,MECKLING K A,MARCONE M F,et al.Can phytochemical antioxidant rich foods act as anti-cancer agents?[J]. Food Research International, 2011, 44(9): 2545-2554. DOI:10.1016/j.foodres.2011.05.021.
[28] SPRANGER I, SUN B S, MATEUS A M et al. Chemical characterization and antioxidant activities of oligomeric and polymeric procyanidin fractions from grape seeds[J]. Food Chemistry, 2008, 108(2): 519-532. DOI:10.1016/j.foodchem.2007.11.004.
[29] FRANKEL E N, WATERHOUSE A L, TEISSEDRE P L. Principal phenolic phytochemicals in selected California wines and their antioxidant activity in inhibiting oxidation of human low-density lipoproteins[J]. Journal of Agricultural and Food Chemistry, 1995, 43(4): 890-894.
[30] ZHANG Shuting, CUI Yan, LI Lingxi, et al. Preparative HSCCC isolation of phloroglucinolysis products from grape seed polymeric proantho-cyani dins as new powerful antioxidants[J]. Food Chemistry, 2015,188:422-429.DOI:10.1016/j.foodchem.2015.05.030.
[31] WANG C C, HUANG P L, LIU T Y, et al. Highly oligomeric procyanidins from areca nut induce lymphocyte apoptosis via the depletion of intracellular thiols[J]. Toxicology in Vitro, 2009, 23(7): 1234-1241.DOI:10.1016/j.tiv.2009.07.033.
[32] MATéS J M, PéREZ-GóMEZ C, Nú?EZ D C I. Antioxidant enzymes and human diseases[J]. Clinical Biochemistry, 1999, 32(8): 595-603.DOI:10.1016/S0009-9120(99)00075-2.
[33]许慧.莲房原花青素诱导ROS积蓄介导HepG2细胞自噬和凋亡的研究[D].镇江:江苏大学,2016:37-39.
[34] LEFRANC F, MIJATOVIC T, KONDO Y, et al. Targeting the α1 subunit of the sodium pump to combat glioblastoma cells[J]. Neurosurgery, 2008, 62(1): 211-222. DOI:10.1227/01.NEU.0000311080.43024.0E.
[35]WINNICKA K,BIELAWSKI K,BIELAWSKA A,et al.Antiproliferative activity of derivatives of ouabain, digoxin and proscillaridina in human MCF-7 and MDA-MB-231 breast cancer cells[J]. Biological and Pharmaceutical Bulletin, 2008, 31(6): 1131-1140.DOI:10.1248/bpb.31.1131.
[36] PAPADEMETRIO D L, TRABUCCHI A, CAVALIERE V, et al. The catechin flavonoid reduces proliferation and induces apoptosis of murine lymphoma cells LB02 through modulation of antiapoptotic proteins[J]. Brazilian Journal of Pharmacognosy, 2013, 23(3): 455-463.DOI:10.1590/S0102-695X2013005000025.
[37] MARTINOU J C, YOULE R J. Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics[J]. Developmental Cell, 2011, 21(1): 92-101. DOI:10.1016/j.devcel.2011.06.017.
[38] MARSDEN V S, O’CONNOR L, O’REILLY L A, et al. Apoptosis initiated by Bcl-2-regulated caspase activation independently of the cytochrome C/Apaf-1/Caspase-9 apoptosome[J]. Nature, 2002, 419: 634-637.DOI:10.1038/nature01101.
[39] OLA M S, NAWAZ M, AHSAN H. Role of Bcl-2 family proteins and caspases in the regulation of apoptosis[J]. Molecular and Cellular Biochemistry, 2011, 351(1/2): 41-58. DOI:10.1007/s11010-010-0709-x.
[40] O U K Q, G U L W. Absorption and metabolism of proanthocyanidins[J].Journal of Functional Foods,2014,7:43-53.DOI:10.1016/j.jff.2013.08.004.
[41] ESPIN J C, LARROSA M, GARCIA-CONESA M T, et al. Biological significance of urolithins, the gut microbial ellagic acid-derived etabolites: the evidence so far[J]. Evidence-Based Complementary and Alternative Medicine,2013,2013:1-15.DOI:10.1155/013/270418.
[2] YANG Y, CHIEN M. Characterization of grape procyanidins using high-performance liquid chromatography/mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry[J]. Journal of Agricultural and Food Chemistry, 2000, 48(9): 3990-3996. DOI:10.1021/jf000316q.
[3] CUI T, LI J Z, KAYAHARA H, et al. Quantification of the polyphenols and triterpene acids in Chinese hawthorn fruit by high-performance liquid chromatography[J]. Journal of Agricultural and Food Chemistry, 2006, 54(13): 4574-4581. DOI:10.1021/jf060310m.
[4] PUIGGRòS F, SALVADò M J, BLADé C, et al. Differential modulation of apoptotic processes by proanthocyanidins as a dietary strategy for delaying chronic pathologies[J]. Critical Reviews in Food Science and Nutrition, 2014, 54(3): 277-291. DOI:10.1080/10408398.2011.565456.
[5]禹华娟,孙智达,谢笔钧.莲原花青素在油脂体系中的抗氧化作用[J].中国农业科学, 2010, 43(10): 2132-2140. DOI:10.3864/j.issn.0578-1752.2010.10.020.
[6]谢文利,晋玉章,万宗明,等.聚果多酚抗衰老作用的实验研究[J].食品科学, 2009, 30(9): 207-209.
[7]PRASANNA R,HARISH C C,PICHAI R,et al.Anti-cancer effect of Cassia auriculata leaf extract in vitro through cell cycle arrest and induction of apoptosis in human breast and larynx cancer cell lines[J]. Cell Biology International, 2009, 33(2): 127-134. DOI:10.1016/j.cellbi.2008.10.006.
[8] GAO J, INAGAKI Y, LIU Y. Research progresses on flavonoids isolated from traditional Chinese medicine in treatment of Alzheimer’s disease[J]. Intractable and Rare Diseases Research, 2013, 2(1): 3-10. DOI:10.5582/irdr.2013.v2.1.3.
[9] WANG L, FUMOTO T, MASUMOTO S, et al. Regression of atherosclerosis with apple procyanidins by activating the ATP-binding cassette subfamily a member 1 in a rabbit model[J]. Atherosclerosis, 2017,258:56-64.DOI:10.1016/j.atherosclerosis.2017.01.032.
[10] LIANG Y, WANG J, GAO H Q, et al. Beneficial effects of grape seed proanthocyanidin extract on arterial remodeling in spontaneously hypertensive rats via protecting against oxidative stress[J]. Molecular Medicine Reports, 2016, 14(4): 3711-3718. DOI:10.3892/mmr.2016.5699.
[11] LIU Z Q, YANG Q L, ZHANG C S, et al. Study on antioxidant activity of proanthocyanidins from peanut skin[J]. Advanced Materials Research,2011,197/198:1582-1586.DOI:10.4028/www.scientific.net/AMR.197-198.1582.
[12]JORD?O A M,GON?A LVES F J,CORREIAA C,et al.Proanthocyanidin content, antioxidant capacity and scavenger activity of Portuguese sparkling wines (Bairrada appellation of origin)[J]. Journal of the Science of Food and Agriculture, 2010, 90(12): 2144-2152.DOI:10.1002/jsfa.4064.
[13] WANG Xinghui, HUANG Lingling, YU Tingting, et al. Effects of oligomeric grape seed proanthocyanidins on heart, aorta, kidney in DOCA-salt mice: role of oxidative stress[J]. Phytotherapy Research, 2013, 27(6): 869-876. DOI:10.1002/ptr.4793.
[14] AN J, CHEN C, WANG X, et al. Oligomeric proanthocyanidins alleviate hexabromocyclododecane nduced cytotoxicity in HepG2 cells through regulation on ROS formation and mitochondrial pathway[J]. Toxicology in Vitro, 2014, 28(2): 319-326. DOI:10.1016/j.tiv.2013.11.009.
[15] FARIA A, CALHAU C, DE FREITAS V, et al. Procyanidins as antioxidants and tumor cell growth modulators[J]. Journal of Agricultural and Food Chemistry, 2006, 54(6): 2392-2397. DOI:10.1021/jf0526487.
[16] CHEN J Y, XU Z, ZHU W, et al. Novel proanthocyanidin dimer analogues with the C-ring-opened diaryl-propan-2-gallate structural unit and enhanced antioxidant activities[J].Journal of Functional Foods,2016,21:290-300.DOI:10.1016/j.jff.2015.12.023.
[17] AGUILERA Y, ESTRELLA I, BENITEZ V, et al. Bioactive phenolic compounds and functional properties of dehydrated bean flours[J]. Food Research International, 2011, 44(3): 774-780. DOI:10.1016/j.foodres.2011.01.004.
[18]赵丹,曾新安,徐中岳.不同聚合度葡萄皮原花青素的抗氧化活性[J].食品与发酵工业, 2012, 38(4): 4104-4108.DOI:10.13995/j.cnki.11-1802/ts.2012.04.009.
[19] LIZARRAGA D, LOZANO C, BRIEDé J J, et al. The importance of polymerization and galloylation for the antiproliferative properties of procyanidin-rich natural extracts[J]. The FEBS Journal, 2007, 274(18): 4802-4811.DOI:10.1111/j.1742-4658.2007.06010.x.
[20] LOZANO C, JULIá L, JIMéNEZ A, et al. Electron-transfer capacity of catechin derivatives and influence on the cell cycle and apoptosis in HT29 cells[J]. The FEBS Journal, 2006, 273(11): 2475-2486. DOI:10.1111/j.1742-4658.2006.05255.x.
[21] BRAY F, FERLAY J, SOERJOMATARAM L, et al. Global Cancer Statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA: A Cancer Journal for Clinicians, 2018, 68(6): 394-424. DOI:10.3322/caac.21492.
[22] CUI T, NAKAMURA K, TIAN S, et al. Polyphenolic content and physiological activities of Chinese hawthorn extracts[J]. Bioscience, Biotechnology, and Biochemistry, 2006, 70(12): 2948-2956. DOI:10.1271/bbb.60361.
[23]谭卫东,金红,罗弟祥,等.抗肿瘤药物筛选中MTT法和SRB法的比较[J].天然产物研究与开发, 1999, 11(3): 17-22. DOI:10.16333/j.1001-6880.1999.03.004.
[24]董成龙,游娜,许向红,等. 白藜芦醇对人甲状腺乳头状癌IHH4细胞增殖和凋亡的影响[J].南京医科大学学报(自然科学版), 2015, 35(8): 1096-1103. DOI:10.7655/NYDXBNS20150809.
[25] ROY A M, BALIGA M S, ELMETS C A, et al. Grape seed proanthocyanidins induce apoptosis through p53, Bax, and Caspase 3 pathways[J]. NeoPlasia, 2005, 7(1): 24-36.DOI:10.1593/neo.04412.
[26]SHAHIDI F,AMBIGAIPALAN P.Phenolics and polyphenolics in foods, beverages and spices: antioxidant activity and health effects: a review[J]. Journal of Functional Foods, 2015, 18: 820-897. DOI:10.1016/j.jff.2015.06.018.
[27]WANG S N,MECKLING K A,MARCONE M F,et al.Can phytochemical antioxidant rich foods act as anti-cancer agents?[J]. Food Research International, 2011, 44(9): 2545-2554. DOI:10.1016/j.foodres.2011.05.021.
[28] SPRANGER I, SUN B S, MATEUS A M et al. Chemical characterization and antioxidant activities of oligomeric and polymeric procyanidin fractions from grape seeds[J]. Food Chemistry, 2008, 108(2): 519-532. DOI:10.1016/j.foodchem.2007.11.004.
[29] FRANKEL E N, WATERHOUSE A L, TEISSEDRE P L. Principal phenolic phytochemicals in selected California wines and their antioxidant activity in inhibiting oxidation of human low-density lipoproteins[J]. Journal of Agricultural and Food Chemistry, 1995, 43(4): 890-894.
[30] ZHANG Shuting, CUI Yan, LI Lingxi, et al. Preparative HSCCC isolation of phloroglucinolysis products from grape seed polymeric proantho-cyani dins as new powerful antioxidants[J]. Food Chemistry, 2015,188:422-429.DOI:10.1016/j.foodchem.2015.05.030.
[31] WANG C C, HUANG P L, LIU T Y, et al. Highly oligomeric procyanidins from areca nut induce lymphocyte apoptosis via the depletion of intracellular thiols[J]. Toxicology in Vitro, 2009, 23(7): 1234-1241.DOI:10.1016/j.tiv.2009.07.033.
[32] MATéS J M, PéREZ-GóMEZ C, Nú?EZ D C I. Antioxidant enzymes and human diseases[J]. Clinical Biochemistry, 1999, 32(8): 595-603.DOI:10.1016/S0009-9120(99)00075-2.
[33]许慧.莲房原花青素诱导ROS积蓄介导HepG2细胞自噬和凋亡的研究[D].镇江:江苏大学,2016:37-39.
[34] LEFRANC F, MIJATOVIC T, KONDO Y, et al. Targeting the α1 subunit of the sodium pump to combat glioblastoma cells[J]. Neurosurgery, 2008, 62(1): 211-222. DOI:10.1227/01.NEU.0000311080.43024.0E.
[35]WINNICKA K,BIELAWSKI K,BIELAWSKA A,et al.Antiproliferative activity of derivatives of ouabain, digoxin and proscillaridina in human MCF-7 and MDA-MB-231 breast cancer cells[J]. Biological and Pharmaceutical Bulletin, 2008, 31(6): 1131-1140.DOI:10.1248/bpb.31.1131.
[36] PAPADEMETRIO D L, TRABUCCHI A, CAVALIERE V, et al. The catechin flavonoid reduces proliferation and induces apoptosis of murine lymphoma cells LB02 through modulation of antiapoptotic proteins[J]. Brazilian Journal of Pharmacognosy, 2013, 23(3): 455-463.DOI:10.1590/S0102-695X2013005000025.
[37] MARTINOU J C, YOULE R J. Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics[J]. Developmental Cell, 2011, 21(1): 92-101. DOI:10.1016/j.devcel.2011.06.017.
[38] MARSDEN V S, O’CONNOR L, O’REILLY L A, et al. Apoptosis initiated by Bcl-2-regulated caspase activation independently of the cytochrome C/Apaf-1/Caspase-9 apoptosome[J]. Nature, 2002, 419: 634-637.DOI:10.1038/nature01101.
[39] OLA M S, NAWAZ M, AHSAN H. Role of Bcl-2 family proteins and caspases in the regulation of apoptosis[J]. Molecular and Cellular Biochemistry, 2011, 351(1/2): 41-58. DOI:10.1007/s11010-010-0709-x.
[40] O U K Q, G U L W. Absorption and metabolism of proanthocyanidins[J].Journal of Functional Foods,2014,7:43-53.DOI:10.1016/j.jff.2013.08.004.
[41] ESPIN J C, LARROSA M, GARCIA-CONESA M T, et al. Biological significance of urolithins, the gut microbial ellagic acid-derived etabolites: the evidence so far[J]. Evidence-Based Complementary and Alternative Medicine,2013,2013:1-15.DOI:10.1155/013/270418.