马齿苋多糖抑制HepG2细胞存活的作用机制
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摘要
探讨马齿苋多糖(polysaccharide of Portulaca oleracea L.,POP-A)对HepG2细胞的抑制作用及作用机制。体外培养HepG2细胞,分别用不同浓度的POP-A处理细胞,采用四甲基偶氮唑盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]法、荧光定性定量法、免疫印迹法以及反转录聚合酶链式反应(reverse transcription-PCR,RT-PCR)法等检测POP-A对HepG2细胞存活率、线粒体膜电位、细胞内钙离子以及相关凋亡蛋白与基因表达的影响。结果表明:当POP-A浓度为1.25、2.5、5 mg/mL时,POP-A能够显著抑制HepG2肝癌细胞、降低线粒体膜电位、增加细胞内的钙离子浓度、提高p53和Bax的基因及蛋白表达。说明POP-A有抑制HepG2肝癌细胞的作用,其作用机制与促进HepG2细胞凋亡有关。
To explore the inhibitory effect and primary mechanism of polysaccharide from Portulaca oleracea L.(POP-A)on growth of HepG2 cells.HepG2 cells were cultured in vitro.The effect of POP-A on cells viability,mitochondrial membrane potential,intracellular Ca2+levels and the expression levels of related apoptotic proteins and gene of HepG2 cells were studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT) assay,fluorescence quantitative analysis,Western blotting and reverse transcription-PCR(RT-PCR).Results showed that 1.25,2.5,5 mg/mL POP-A significantly inhibited the cell viability of HepG2 cells,decreased mitochondrial membrane potential,increased intracellular Ca2+levels and increased the gene and protein expression levels of p53 and Bax.These results suggested that POP-A could inhibit the apoptosis of HepG2 cells,and its mechanism was related to the promotion of apoptosis of HepG2 cells.
To explore the inhibitory effect and primary mechanism of polysaccharide from Portulaca oleracea L.(POP-A)on growth of HepG2 cells.HepG2 cells were cultured in vitro.The effect of POP-A on cells viability,mitochondrial membrane potential,intracellular Ca2+levels and the expression levels of related apoptotic proteins and gene of HepG2 cells were studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT) assay,fluorescence quantitative analysis,Western blotting and reverse transcription-PCR(RT-PCR).Results showed that 1.25,2.5,5 mg/mL POP-A significantly inhibited the cell viability of HepG2 cells,decreased mitochondrial membrane potential,increased intracellular Ca2+levels and increased the gene and protein expression levels of p53 and Bax.These results suggested that POP-A could inhibit the apoptosis of HepG2 cells,and its mechanism was related to the promotion of apoptosis of HepG2 cells.
引文
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[2]WANG P,SUN H,LIU D,et al.Protective effect of a phenolic extract containing indoline amides from Portulaca oleracea against cognitive impairment in senescent mice induced by large dose of D-galactose/NaNO2[J].Journal of Ethnopharmacol,2017,203:252-259
[3]OH K B,CHANG I M,HWANG K J,et al.Detection of antifungal activity in Portulaca oleracea by a single-cell bioassay system[J].Phytotherapy Research,2000,14(5):329-332
[4]DEHGHAN F,Soori R,Gholami K,et al.Purslane(Portulaca oleracea)Seed Consumption And Aerobic Training Improves Biomarkers Associated with Atherosclerosis in Women with Type 2 Diabetes(T2D)[J].Sciences Report,2016,6:37819
[5]BAI Y,ZANG X,MA J,et al.Anti-Diabetic Effect of Portulaca oleracea L.Polysaccharideandits Mechanism in Diabetic Rats[J].International Journal of Molecular Sciences,2016,17(8):1201
[6]GUO G,YUE L,FAN S,et al.Antioxidant and Antiproliferative Activities of Purslane Seed Oil[J].Journal of hypertension:open access,2016,5(2):218
[7]LI C Y,MENG Y H,YING Z M,et al.Correction to Three Novel Alkaloids from Portulaca oleracea L.and Their Anti-inflammatory Effects[J].Journal of Agric Food Chemical,2017,65(4):993-994
[8]LEE A S,KIM J S,LEE Y J,et al.Anti-TNF-αActivity of Portulaca oleracea in Vascular Endothelial Cells[J].International Journal of Molecular Sciences,2012,13(5):5628-5644
[9]ASKARI V R,REZAEE S A,ABNOUS K,et al.The influence of hydro-ethanolic extract of Portulaca oleracea L.on Th1/Th2 balance in isolated human lymphocytes[J].Journal of Ethnopharmacology,2016,194:1112-1121
[10]ZHAO R,ZHANG T,MA B,et al.Antitumor Activity of Portulaca oleracea L.Polysaccharide on He La Cells Through Inducing TLR4/NF-κB Signaling[J].Nutrition and cancer,2017,69(1):131-139
[11]翁前锋,袁凯龙,张宏颖,等.胶束电动毛细管色谱安培检测中药马齿苋中多巴胺和去甲肾上腺素[J].色谱,2005,23(1):18-21
[12]YIN A W,GAO P F,DANG L M.Anti-aging Effects of Flavonoids from Portulaca oleracea L.[J].Natural Product Research and Development,2017,29(6):988-993
[13]汪荔,王征,张娇,等.传统工艺与膜分离技术联合对马齿苋多糖的提取分离与抗氧化活性研究[J].中草药,2016,47(10):1676-1681
[14]郑智音,贾晓斌,舒娈,等.鲜马齿苋多糖、生物碱和多酚类组分的制备及其降血糖活性研究[J].中草药,2014,45(18):2673-2677
[15]ZHAO C,YING Z,TAO X,et al.A new lactam alkaloid from Portulaca oleracea L.and its cytotoxity[J].Natural Product Research,2017:1-6
[16]Li Y P,YAO L H,WU G J,et al.Antioxidant activities of novel small-molecule polysaccharide fractions purified from Portulaca oleracea L.[J].Food Science and Biotechnology,2014,23(6):2045-2052
[17]ZHAO R,ZHANG T,ZHAO H,et al.Effects of Portulaca oleracea L.Polysaccharides on Phenotypic and Functional Maturation of Murine Bone Marrow Derived Dendritic Cells[J].Nutrition&Cancer,2015,67(6):987-993
[18]ZHAO R,GAO X,CAI Y,et al.Antitumor activity of Portulaca oleracea L.polysaccharides against cervical carcinoma in vitro and in vivo[J].Carbohydrate polymers,2013,96(2):376-383
[19]SHEN H,GUO T,ZENG G,et al.Purification and characterization of an antitumor polysaccharide from Portulaca oleracea L.[J].Carbohydr Polym,2013,93(2):395-400
[20]GONG F,LI F,ZHANG L,et al.Hypoglycemic effects of crude polysaccharide from Purslane[J].International Journal of Molecular Sciences,2009,10(3):880-888
[21]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作用[J].食品科学,2017,38(3):201-205
[22]XIAO J L,YONG H,XUE Q G,et al.Polysaccharides from Portulaca oleracea(purslane)supplementation lowers acute exercise induced oxidative stress in young rats[J].African Journal of Pharmacy and Pharmacology,2011,5(3):381-385
[23]梁彦,吕艳荣.马齿苋多糖的抗衰老作用[J].江苏农业科学,2014,42(4):270-272
[24]崔旻,尹苗,安利国.马齿苋多糖的抗肿瘤活性[J].山东师范大学学报(自然科学版),2002,17(1):72-76
[25]王晓波,姜红,王本华,等.马齿苋多糖对肿瘤细胞的体内外抑制作用[J].中国公共卫生,2005,21(12):1485-1486
[26]丁虹,唐雯静,庞燕芬.马齿苋多糖对宫颈癌细胞恶性生物学行为抑制作用的实验研究[J].东南国防医药,2016,18(1):38-40,58
[27]胡庆娟,吴光杰,牛庆川,等.响应面法优化木瓜蛋白酶法脱马齿苋多糖蛋白工艺[J/OL].食品科学.http://kns.cnki.net/kcms/detail/11.2206.TS.20180209.1041.078.html
[28]UEDA M,ASHIDA H.Green Tea Prevents Obesity by Increasing Expression of Insulin-like Growth Factor Binding Protein-1 in Adipose Tissue of High-Fat Diet-Fed Mice[J].Journal of Agricultural and Food Chemistry,2012,60(36):8917-8923
[29]张贤益,李文娟,钟亮,等.白扁豆多糖对神经细胞缺氧性凋亡的保护机制[J].食品科学,2018,39(3):222-228
[30]GHOBRIAL I M,WITZIG T E,ADJEI A A,et al.Targeting Apoptosis Pathways in Cancer Therapy[J].CA:A Cancer Journal for Clinicians,2005,55(3):178-194
[31]WONG R S.Apoptosis in cancer:from pathogenesis to treatment[J].Journal of Experimental&Clinical Cancer Research,2011,30(1):87-87