马齿苋结合态多酚和自由态多酚抑制肝癌细胞HepG-2增殖及诱导细胞凋亡的研究
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摘要
[目的]研究马齿苋结合态多酚和自由态多酚抑制肝癌细胞HepG-2增殖并诱导细胞凋亡的作用。[方法]将不同浓度马齿苋结合态多酚和自由态多酚与HepG-2细胞在体外培养,采用MTT检测细胞增殖活性;光镜观察细胞形态;流式细胞术分别检测细胞周期、细胞凋亡和线粒体膜电位;酶标仪检测Caspase酶活力。[结果]结果表明,马齿苋多酚对肝癌细胞HepG-2的增殖抑制作用和凋亡促进作用与一定范围的处理浓度和处理时间呈正相关,36h、0.4g·L-1处理条件下,马齿苋自由态多酚对HepG-2细胞的抑制率仅为15.7%,而马齿苋结合态多酚对HepG-2细胞的抑制率达到73.8%;马齿苋结合态多酚作用下,HepG-2肝癌细胞光镜下可观察到细胞出现明显的形态学改变,处理后细胞周期阻滞于S期,Caspase-3、Caspase-8、Caspase-9酶活性也显著提高,并诱导细胞发生凋亡。马齿苋结合态多酚可能通过激活膜受体通路和线粒体介导的内源性凋亡通路诱导HepG-2细胞凋亡。
[Objective]The present study was undertaken to analyze the induced apoptosis and proliferation inhibition influence of bound polyphenoland freepolyphenol in Portulaca Oleracea L.to HepG-2 liver cancer cells.[Methods]Different concentration of the bound polyphenol in Portulaca Oleracea L.(BPPO)and free polyphenol in Portulaca Oleracea L.(FPPO)was used to in vitro cultured with HepG-2 liver cancer cell line.The cell proliferation activity was analyzed by MTT;the cell morphology was observed by light microscope;the cell cycle,apoptosis and mitochondrial membrane potential assay were analyzed by flow cytometry;and the caspases enzymatic activity assay was analyzed by microplate reader.[Conclusion]The result showed that BPPO significantly inhibited the proliferation of HepG-2 cells in a concentration-and time-dependent manner.Significant morphological change in cells could be observed under the microscope.After treatment with BPPO,the cell cycle of HepG-2 cells was stopped in S phase,and the enzyme activity of caspase 3,caspase 8 and caspase 9 were also significantly increased.In summary,the BPPO could significantly inhibit the proliferation and induce apoptosis of HepG-2 cells in a dose-and time-dependent manner,and might induce apoptosis of HepG2 cells by activating membrane receptor pathway and mitochondrial pathway.
[Objective]The present study was undertaken to analyze the induced apoptosis and proliferation inhibition influence of bound polyphenoland freepolyphenol in Portulaca Oleracea L.to HepG-2 liver cancer cells.[Methods]Different concentration of the bound polyphenol in Portulaca Oleracea L.(BPPO)and free polyphenol in Portulaca Oleracea L.(FPPO)was used to in vitro cultured with HepG-2 liver cancer cell line.The cell proliferation activity was analyzed by MTT;the cell morphology was observed by light microscope;the cell cycle,apoptosis and mitochondrial membrane potential assay were analyzed by flow cytometry;and the caspases enzymatic activity assay was analyzed by microplate reader.[Conclusion]The result showed that BPPO significantly inhibited the proliferation of HepG-2 cells in a concentration-and time-dependent manner.Significant morphological change in cells could be observed under the microscope.After treatment with BPPO,the cell cycle of HepG-2 cells was stopped in S phase,and the enzyme activity of caspase 3,caspase 8 and caspase 9 were also significantly increased.In summary,the BPPO could significantly inhibit the proliferation and induce apoptosis of HepG-2 cells in a dose-and time-dependent manner,and might induce apoptosis of HepG2 cells by activating membrane receptor pathway and mitochondrial pathway.
引文
[1]Sun X,Li L,Ma H G,et al.Bisindolylmaleimide alkaloid BMA-155Cl induces autophagy and apoptosis in human hepatocarcinoma HepG-2cells through the NF-κB p65pathway[J].Acta Pharmacol Sin,2017,38(4):524-538.
[2]田淋,莫发荣.多免疫途径对肝癌治疗的研究进展[J].中国医院药学杂志,2018,38(8):897-900.
[3]邢雪.肝脏恶性肿瘤外科治疗进展[J].国际外科学杂志,1992(3):132-135
[4]汤勇,周训平,王晓芹.原发性肝癌的基因治疗进展[J].局解手术学杂志,2009,18(3):202-203.
[5]潘扬勋,陈锦滨,陈健聪,等.单孔腹腔镜在肝癌治疗的安全性及可行性研究[J].中华腔镜外科杂志,2018,11(1):47-50.
[6]张厚云,陈书凯,李学敏,等.射频消融、高频热疗联DC/CIK细胞治疗中晚期肝癌的临床研究[J].临床和实验医学杂志,2018(2):287-290.
[7]丁怀伟,姚佳琪,宋少江.马齿苋的化学成分和药理活性研究进展[J].沈阳药科大学学报,2008,25(10):831-838.
[8]牛广财,李世燕,朱丹,等.马齿苋多糖POPⅡ和POPⅢ的抗肿瘤及提高免疫力的作用[J].食品科学,2017,38(3):201-205.
[9]尹爱武,高鹏飞,党丽敏.马齿苋黄酮抗衰老作用研究(英文)[J].天然产物研究与开发,2017(6):92-97.
[10]Lei X,Li J,Liu B,et al.Separation and identification of four new compounds with antibacterial activity fromPortulaca oleracea L.[J].Molecules,2015,20(9):16375-16387.
[11]Martins W B,Rodrigues S A,Silva H K,et al.Neuroprotective effect of Portulaca oleracea extracts against 6-hydroxydopamine-induced lesion of dopaminergic neurons[J].An Acad Bras Cienc,2016,88(3):1439-1450.
[12]Bai Y,Zang X,Ma J,et al.Anti-diabetic effect of Portulaca oleracea L.polysaccharideandits mechanism in diabetic rats[J].Int J Mol Sci,2016,17(8):1201.
[13]王天宁,刘玉婷,肖凤琴,等.马齿苋的化学成分和药理活性的现代研究研究整理[J].中国实验方剂学杂志,2018(6):224-234.
[14]Jin T,Shen T,Zhou M,et al.Chemical constituents fromPortulaca oleracea and their bioactivities[J].J Chin Pharmacol Sci,2016,25(12):898-905.
[15]Szliszka E,Krol W.Polyphenols isolated from propolis augment trail-induced apoptosis in cancer cells[J].Evid Based Complement Alternat Med,2013,2013:1-10.
[16]于萍.葡多酚对乳腺癌抑制作用的体内外研究[D].青岛:青岛大学,2007.
[17]Shuhua S,Yue X,Zhao H L.Bound polyphenol extracted from jujube pulp triggers mitochondriamediated apoptosis and cell cycle arrest of HepG-2cell in vitro and in vivo[J].J Funct Foods,2018,53(2019):187-196.
[18]史江颖,单树花,李宗伟,等.谷糠结合态多酚对四种肿瘤细胞增殖的抑制作用[J].营养学报,2015,37(2):178-184.
[19]Zhao R,Gao X,Cai Y,et al.Antitumor activity of Portulaca oleracea L.polysaccharides against cervical carcinoma in vitro and in vivo[J].Carbohyd Polym,2013,96(2):376-383.
[20]Shen H,Tang G,Zeng G,et al.Purification and characterization of an antitumor polysaccharide fromPortulaca oleracea L[J].Carbohyd Polym,2013,93(2):395-400.
[21]吴诗玄.龙眼中多酚的提取、分离纯化、结构鉴定及体外抗氧化活性测定[D].广州:暨南大学,2018.
[22]Yang Y,Zhang T.Antimicrobial activities of tea polyphenol on phytopathogens:a review[J].Molecules,2019,24(816):1-8.
[23]董丽华,杨鹏春,戴迟兵.茶多酚对人结肠癌细胞株Caco-2细胞凋亡及细胞周期的影响[J].广东医学,2015,36(1):50-52.
[24]高晓天,容秋萍,宋泽庆.白藜芦醇诱导肺癌细胞凋亡和细胞周期阻滞的分子机制[J].中国热带医学,2017,17(4):350-354.
[25]章迅,吴艾平,章永红.石榴皮多酚对人乳腺癌细胞MDA-MB-231增殖及凋亡的影响[J].天津中医药大学学报,2012,31(4):214-217.
[26]詹玮,田甜,余蕾,等.蓝莓花青素对HepG-2细胞凋亡及组蛋白乙酰化修饰的影响[J].中国普通外科杂志,2018,27(1):55-60.
[27]杨涛,费振海,钟兴明.Caspase家族与细胞凋亡的研究进展[J].浙江医学,2018,40(18):2083-2087.
[28]李萍,杨志刚,文国容,等.茶多酚对ACC-M细胞株Fas、Fas l表达的影响[J].重庆医学,2010,39(3):1219-1221.
[29]Gupta K,Thakur V S,Bhaskaran N,et al.Green Tea Polyphenols Induce p53-Dependent and p53-Independent Apoptosis in Prostate Cancer Cells through Two Distinct Mechanisms[J].PLoS One,2012,7(12).
[30]石静,卿晨.线粒体膜电位改变与细胞凋亡[J].中国民族民间医药,2011,20(7):20-21.
[31]陈慧莉,李建华,王树庆.线粒体跨膜电位和细胞凋亡相关性的研究[J].医学综述,2007(14):1041-1043.
[2]田淋,莫发荣.多免疫途径对肝癌治疗的研究进展[J].中国医院药学杂志,2018,38(8):897-900.
[3]邢雪.肝脏恶性肿瘤外科治疗进展[J].国际外科学杂志,1992(3):132-135
[4]汤勇,周训平,王晓芹.原发性肝癌的基因治疗进展[J].局解手术学杂志,2009,18(3):202-203.
[5]潘扬勋,陈锦滨,陈健聪,等.单孔腹腔镜在肝癌治疗的安全性及可行性研究[J].中华腔镜外科杂志,2018,11(1):47-50.
[6]张厚云,陈书凯,李学敏,等.射频消融、高频热疗联DC/CIK细胞治疗中晚期肝癌的临床研究[J].临床和实验医学杂志,2018(2):287-290.
[7]丁怀伟,姚佳琪,宋少江.马齿苋的化学成分和药理活性研究进展[J].沈阳药科大学学报,2008,25(10):831-838.
[8]牛广财,李世燕,朱丹,等.马齿苋多糖POPⅡ和POPⅢ的抗肿瘤及提高免疫力的作用[J].食品科学,2017,38(3):201-205.
[9]尹爱武,高鹏飞,党丽敏.马齿苋黄酮抗衰老作用研究(英文)[J].天然产物研究与开发,2017(6):92-97.
[10]Lei X,Li J,Liu B,et al.Separation and identification of four new compounds with antibacterial activity fromPortulaca oleracea L.[J].Molecules,2015,20(9):16375-16387.
[11]Martins W B,Rodrigues S A,Silva H K,et al.Neuroprotective effect of Portulaca oleracea extracts against 6-hydroxydopamine-induced lesion of dopaminergic neurons[J].An Acad Bras Cienc,2016,88(3):1439-1450.
[12]Bai Y,Zang X,Ma J,et al.Anti-diabetic effect of Portulaca oleracea L.polysaccharideandits mechanism in diabetic rats[J].Int J Mol Sci,2016,17(8):1201.
[13]王天宁,刘玉婷,肖凤琴,等.马齿苋的化学成分和药理活性的现代研究研究整理[J].中国实验方剂学杂志,2018(6):224-234.
[14]Jin T,Shen T,Zhou M,et al.Chemical constituents fromPortulaca oleracea and their bioactivities[J].J Chin Pharmacol Sci,2016,25(12):898-905.
[15]Szliszka E,Krol W.Polyphenols isolated from propolis augment trail-induced apoptosis in cancer cells[J].Evid Based Complement Alternat Med,2013,2013:1-10.
[16]于萍.葡多酚对乳腺癌抑制作用的体内外研究[D].青岛:青岛大学,2007.
[17]Shuhua S,Yue X,Zhao H L.Bound polyphenol extracted from jujube pulp triggers mitochondriamediated apoptosis and cell cycle arrest of HepG-2cell in vitro and in vivo[J].J Funct Foods,2018,53(2019):187-196.
[18]史江颖,单树花,李宗伟,等.谷糠结合态多酚对四种肿瘤细胞增殖的抑制作用[J].营养学报,2015,37(2):178-184.
[19]Zhao R,Gao X,Cai Y,et al.Antitumor activity of Portulaca oleracea L.polysaccharides against cervical carcinoma in vitro and in vivo[J].Carbohyd Polym,2013,96(2):376-383.
[20]Shen H,Tang G,Zeng G,et al.Purification and characterization of an antitumor polysaccharide fromPortulaca oleracea L[J].Carbohyd Polym,2013,93(2):395-400.
[21]吴诗玄.龙眼中多酚的提取、分离纯化、结构鉴定及体外抗氧化活性测定[D].广州:暨南大学,2018.
[22]Yang Y,Zhang T.Antimicrobial activities of tea polyphenol on phytopathogens:a review[J].Molecules,2019,24(816):1-8.
[23]董丽华,杨鹏春,戴迟兵.茶多酚对人结肠癌细胞株Caco-2细胞凋亡及细胞周期的影响[J].广东医学,2015,36(1):50-52.
[24]高晓天,容秋萍,宋泽庆.白藜芦醇诱导肺癌细胞凋亡和细胞周期阻滞的分子机制[J].中国热带医学,2017,17(4):350-354.
[25]章迅,吴艾平,章永红.石榴皮多酚对人乳腺癌细胞MDA-MB-231增殖及凋亡的影响[J].天津中医药大学学报,2012,31(4):214-217.
[26]詹玮,田甜,余蕾,等.蓝莓花青素对HepG-2细胞凋亡及组蛋白乙酰化修饰的影响[J].中国普通外科杂志,2018,27(1):55-60.
[27]杨涛,费振海,钟兴明.Caspase家族与细胞凋亡的研究进展[J].浙江医学,2018,40(18):2083-2087.
[28]李萍,杨志刚,文国容,等.茶多酚对ACC-M细胞株Fas、Fas l表达的影响[J].重庆医学,2010,39(3):1219-1221.
[29]Gupta K,Thakur V S,Bhaskaran N,et al.Green Tea Polyphenols Induce p53-Dependent and p53-Independent Apoptosis in Prostate Cancer Cells through Two Distinct Mechanisms[J].PLoS One,2012,7(12).
[30]石静,卿晨.线粒体膜电位改变与细胞凋亡[J].中国民族民间医药,2011,20(7):20-21.
[31]陈慧莉,李建华,王树庆.线粒体跨膜电位和细胞凋亡相关性的研究[J].医学综述,2007(14):1041-1043.
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