蓝莓花色苷提取物干预人肝癌细胞对葡萄糖消耗的作用研究
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摘要
探究蓝莓花色苷提取物对胰岛素抵抗的人肝癌细胞HepG2细胞对葡萄糖消耗的干预作用。利用MTT实验筛选得到3个品种蓝莓花色苷提取物的最适作用浓度;筛选胰岛素和葡萄糖诱导HepG2细胞产生胰岛素抵抗的最佳浓度;以MTT矫正细胞数量,用葡萄糖氧化酶法检测细胞培养基中葡萄糖的变化来研究蓝莓花色苷提取物的对细胞胰岛素抵抗的干预作用。北陆花色苷提取物在低于200μg/m L的浓度时,灿烂和园蓝花色苷提取物在低于100μg/m L的浓度时,对HepG2细胞活力无明显影响;以诱导剂胰岛素的浓度为1×10-8mol/L,孵育24h作为诱导条件,其模型效果较佳、对细胞活力改变小且稳定性好;3个品种的蓝莓花色苷提取物在7.8125～31.25μg/m L浓度时,可不同程度促进正常HepG2细胞和胰岛素抵抗的HepG2细胞对葡萄糖的消耗量,且与剂量成正比关系,在31.25μg/m L达到促进最大值,与模型组比较均有极显著差异,糖消耗量分别增加了60%、65%和88%。三个品种的蓝莓花色苷提取物均能较好的预防和改善胰岛素抵抗HepG2细胞对葡萄糖的利用。
The regulation of glucose consumption in insulin-resistant hepatoma cell line HepG2 with blueberry anthocyanin extracts was investigated. The optimum concentration of blueberry anthocyanin extracts was screened by MTT assay. Different concentrations of insulin and glucose were performed to induce insulin resistance in HepG2 cells. The number of cells was corrected by MTT, and contents of glucose in medium were detected by glucose oxidase method. Changes in glucose in the medium reflected the intervention of the blueberry anthocyanin extract on cellular insulin resistance. MTT results showed that and there was no significant effect on the activity of HepG2 cells at concentrations below 200 μg/m L of the extracts of NL BAEs and that below 100 μg/m L of the extracts of BW BAEs and GB BAEs. With the inducing agent insulin concentration of 1×10-8 mol/L and incubation for 24 h as the induction condition, the model was effective, while the cell viability had no significant change and the stability was good. It could promote glucose consumption in normal HepG2 cells and insulin-resistant HepG2 cells at the concentration of 7.8125～31.25 μg/m L of three varieties of blueberry anthocyanin extracts and showed a dose-effect relationship. It had a maximum at 31.25 μg/m L. Compared to the model group, sugar consumptions of extracts of NL BAEs, BW BAEs and GB BAEs had increased by 60%, 65% and 88%, respectively. Three varieties of blueberry anthocyanin extract could prevent and improve the utilization of glucose by insulin-resistant HepG2 cells.
The regulation of glucose consumption in insulin-resistant hepatoma cell line HepG2 with blueberry anthocyanin extracts was investigated. The optimum concentration of blueberry anthocyanin extracts was screened by MTT assay. Different concentrations of insulin and glucose were performed to induce insulin resistance in HepG2 cells. The number of cells was corrected by MTT, and contents of glucose in medium were detected by glucose oxidase method. Changes in glucose in the medium reflected the intervention of the blueberry anthocyanin extract on cellular insulin resistance. MTT results showed that and there was no significant effect on the activity of HepG2 cells at concentrations below 200 μg/m L of the extracts of NL BAEs and that below 100 μg/m L of the extracts of BW BAEs and GB BAEs. With the inducing agent insulin concentration of 1×10-8 mol/L and incubation for 24 h as the induction condition, the model was effective, while the cell viability had no significant change and the stability was good. It could promote glucose consumption in normal HepG2 cells and insulin-resistant HepG2 cells at the concentration of 7.8125～31.25 μg/m L of three varieties of blueberry anthocyanin extracts and showed a dose-effect relationship. It had a maximum at 31.25 μg/m L. Compared to the model group, sugar consumptions of extracts of NL BAEs, BW BAEs and GB BAEs had increased by 60%, 65% and 88%, respectively. Three varieties of blueberry anthocyanin extract could prevent and improve the utilization of glucose by insulin-resistant HepG2 cells.
引文
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[25]张卓睿,毛迪锐,高晗,等.蓝莓花青素对小鼠抗疲劳及体内抗氧化作用[J].食品科学,2017,38(21):207-211ZHANG Zhuo-rui,MAO Di-rui,GAO Han,et al.Anti-fatigue and antioxidation of blueberry anthocyanins in mice[J].Food Science,2017,38(21):207-211
[26]Nilgun I,Ozan G.Investigation of the impact of blueberries on metabolic factors influencing health[J].Journal of Functional Foods,2017,38(Part A):298-307
[27]Li D,Zhang Y,Liu Y,et al.Purified anthocyanin supplementation reduces dyslipidemia,enhances antioxidant capacity,and prevents insulin resistance in diabetic patients[J].Journal of Nutrition,2015,145(4):742-748
[28]Shi M,Loftus H,Mc Ainch A J,et al.Blueberry as a source of bioactive compounds for the treatment of obesity,type 2diabetes and chronic inflammation[J].Journal of Functional Foods,2017,30:16-29
[2]Liu S X,Yang H Y,Li S Y,et al.Polyphenolic compositions and chromatic characteristics of bog bilberry syrup wines[J].Molecules,2015,20(11):19865-19877
[3]Wu Y,Zhou Q,Chen X Y,et al.Comparison and screening of bioactive phenolic compounds in different blueberry cultivars:Evaluation of anti-oxidation andα-glucosidase inhibition effect[J].Food Research International,2017,100(Part 1):312-324
[4]徐玉涛,李珂珂,王贺新,等.高效液相色谱法对蓝莓果实中8个有机酸含量的测定[J].食品科学,2015,36(18):127-131XU Yu-tao,LI Ke-ke,WANG He-xin,et al.Determination of8 organic acids in blueberry fruit by high performance liquid chromatography[J].Food Science,2015,36(18):127-131
[5]Tian Y,Liimatainen J,Alanne A L,et al.Phenolic compounds extracted by acidic aqueous ethanol from berries and leaves of different berry plants[J].Food Chemistry,2017,220(4):266-281
[6]Musso G,Gambino R,Cassader M.Non-alcoholic fatty liver disease from pathogenesis to management:an update[J].Obesity Reviews,2009,11:430-445
[7]Hajiaghaalipour F,Khalilpourfarshbafi M,Arya A.Modulation of glucose transporter protein by dietary flavonoids in type 2 diabetes mellitus[J].International Journal of Biological Sciences,2018,11(5):508-524
[8]Stull A.Blueberries’impact on insulin resistance and glucose intolerance[J].Antioxidants,2016,5(4)
[9]Yan F,Dai G,Zheng X.Mulberry anthocyanin extract ameliorates insulin resistance by regulating PI3K/AKTpathway in Hep G2 cells and db/db mice[J].The Journal of Nutritional Biochemistry,2016,36(10):68-80
[10]Yan F,Zhang J,Zhang L,et al.Mulberry anthocyanin extract regulates glucose metabolism by promotion of glycogen synthesis and reduction of gluconeogenesis in human Hep G2cells[J].Food&Function,2016,7(1):425-433
[11]黄明辉,葛长杰,于淑娟.山茱萸中马钱苷和莫诺苷对Hep G2细胞及其胰岛素抵抗模型的影响研究[J].现代食品科技,2017,33(4):13-18HUANG Ming-hui,GE Chang-jie,YU Shu-juan.Effects of loganin and morroniside extracted from cornus officinalis on Hep G2 cells and their insulin resistance model[J].Modern Food Science and Technology,2017,33(4):13-18
[12]Nile S H,Park S W.Edible berries:bioactive components and their effect on human health[J].Nutrition,2014,30(2):134-144
[13]Huang H P,Chang Y C,Wu C H,et al.Anthocyanin-rich Mulberry extract inhibit the gastric cancer cell growth in vitro and xenograft mice by inducing signals of p38/p53 and c-jun[J].Food Chemistry,2011,129(4):1703-1709
[14]Wang H,Guo X,Hu X,et al.Comparison of phytochemical profiles,antioxidant and cellular antioxidant activities of different varieties of blueberry(Vaccinium spp.)[J].Food Chemistry,2017,217(C):773-781
[15]Pedro A C,Granato D,Rosso N D.Extraction of anthocyanins and polyphenols from black rice(Oryza sativa L.)by modeling and assessing their reversibility and stability[J].Food Chemistry,2016,191:12
[16]Sun L Q,Ding X P,Qi J,et al.Antioxidant anthocyanins screening through spectrum effect relationships and DPPH-HPLC-DAD analysis on nine cultivars of introduced rabbiteye blueberry in China[J].Food Chemistry,2012,132(2):759-765
[17]Vendrame S,Daugherty A,Kristo AS,et al.Wild blueberry(Vaccinium angustifolium)consumption improves inflammatory status in the obese Zucker rat model of the metabolic syndrome[J].The Journal of Nutritional Biochemistry,2013,24(8):1508-1512
[18]韩彩静,程安玮,陈相艳,等.蓝莓多酚对巨噬细胞中i NOS、COX-2基因表达的影响[J].食品科学,2014,35(19):144-148HAN Cai-jing,CHENG An-wei,CHEN Xiang-yan,et al.Effects of blueberry polyphenols on the expression of i NOSand COX-2 in macrophages[J].Food Science,2014,35(19):144-148
[19]郭红辉,胡艳,刘驰,等.黑米花色苷对果糖喂养大鼠胰岛素敏感性影响[J].中国公共卫生,2008,24(10):1200-1202GUO Hong-hui,HU Yan,LIU Chi,et al.Effect of anthocyanin extracted from black rice on insulin sensitivity in fructose-fed rats[J].Chin J Public Health,2008,24(10):1200-1202
[20]阎芙洁.桑葚花色苷对糖代谢的调控作用及其机制研究[D].杭州:浙江大学,2018YAN Fu-jie.Study on the beneficial effect and mechanism of mulberry anthocyanins on glucose metabolism regulation[D].Hangzhou:Zhejiang University,2018
[21]胡昕倩,柯敏.花色苷对高糖高胰岛素培养下Müller细胞的保护[J].武汉大学学报(医学版),2011,32(5):623-626HU Xin-qian,KE Min.Effect of cyanin in protecting cultured müller cells under high glucose and high insulin condition[J].Medical Journal of Wuhan University,2011,32(5):623-626
[22]Mokashi P,Khanna A,Pandita N.Flavonoids from Enicostema littorale blume enhances glucose uptake of cells in insulin resistant human liver cancer(Hep G2)cell line via IRS-1/PI3K/Akt pathway[J].Biomedicine et Pharmacotherapy,2017,90:268-277
[23]武雪玲,李筱筱,贾世亮,等.黑果枸杞花青素对Aβ42致痴呆模型大鼠记忆力及抗氧化活性研究[J].现代食品科技,2017,33(3):29-34WU Xue-ling,LI Xiao-xiao,JIA Shi-liang,et al.Memory enhancing and antioxidant activities of lycium ruthenicum murray anthocyanin extracts in an Aβ42-induced rat model of dementia[J].Modern Food Science and Technology,2017,33(3):29-34
[24]Vendrame S,Daugherty A,Kristo AS,et al.Wild blueberry(Vaccinium angustifolium)consumption improves inflammatory status in the obese Zucker rat model of the metabolic syndrome[J].The Journal of Nutritional Biochemistry,2013,24(8):1508-1512
[25]张卓睿,毛迪锐,高晗,等.蓝莓花青素对小鼠抗疲劳及体内抗氧化作用[J].食品科学,2017,38(21):207-211ZHANG Zhuo-rui,MAO Di-rui,GAO Han,et al.Anti-fatigue and antioxidation of blueberry anthocyanins in mice[J].Food Science,2017,38(21):207-211
[26]Nilgun I,Ozan G.Investigation of the impact of blueberries on metabolic factors influencing health[J].Journal of Functional Foods,2017,38(Part A):298-307
[27]Li D,Zhang Y,Liu Y,et al.Purified anthocyanin supplementation reduces dyslipidemia,enhances antioxidant capacity,and prevents insulin resistance in diabetic patients[J].Journal of Nutrition,2015,145(4):742-748
[28]Shi M,Loftus H,Mc Ainch A J,et al.Blueberry as a source of bioactive compounds for the treatment of obesity,type 2diabetes and chronic inflammation[J].Journal of Functional Foods,2017,30:16-29