超声-闪式联合法制备蓝莓花色苷提取物及其体内外抗肿瘤活性评价

doi:10.7506/spkx1002-6630-20190411-159

食品科学 ›› 2020, Vol. 41 ›› Issue (6): 259-269.doi: 10.7506/spkx1002-6630-20190411-159

超声-闪式联合法制备蓝莓花色苷提取物及其体内外抗肿瘤活性评价

薛宏坤，谭佳琪，刘成海，刘钗

（1.东北农业大学工程学院，黑龙江哈尔滨 150030；2.东北农业大学农学院，黑龙江哈尔滨 150030）

出版日期:2020-03-25 发布日期:2020-03-23
基金资助:
东北农业大学“青年才俊”项目（518076）

Preparation of Blueberry Anthocyanin Extract by Ultrasound-Assisted Flash Extraction and Evaluation of Its Anti-Tumor Activity in Vivo and in Vitro

XUE Hongkun, TAN Jiaqi, LIU Chenghai, LIU Chai

(1. College of Engineering, Northeast Agricultural University, Harbin 150030, China;2. College of Agricultural, Northeast Agricultural University, Harbin 150030, China）

Online:2020-03-25 Published:2020-03-23

摘要/Abstract

摘要： 以蓝莓为原料，采用超声-闪式联合法提取花色苷，探究超声功率、闪提转速、提取时间、料液比和乙醇体积分数对花色苷含量的影响。在单因素试验的基础上，利用遗传算法优化超声-闪式联合法提取蓝莓花色苷的工艺参数。体外实验采用MTT法和Annexin V-FITC/PI双标记染色法探究不同质量浓度花色苷提取物对S180细胞增殖和凋亡的影响；体内实验利用S180荷瘤小鼠模型和酶联免疫吸附测定法探究花色苷提取物的抗肿瘤活性及其机制。结果表明：超声-闪式联合法提取蓝莓中花色苷的优化工艺参数为提取时间10 min、超声功率450 W、闪提转速8 400 r/min、料液比1∶30（g/mL）和乙醇体积分数68%，在此条件下花色苷含量和纯度分别为（2.71±0.08）mg/g和（31.79±0.53）%。体外实验发现花色苷提取物能有效抑制S180细胞增殖，诱导S180细胞凋亡，使细胞阻滞于S期。荷瘤小鼠体内抑制肿瘤实验表明：花色苷提取物低、中、高剂量组瘤体质量显著低于模型对照组（P＜0.05），抑瘤率分别为26.36%、33.33%、46.51%；花色苷提取物低、中、高剂量组荷瘤小鼠血清中的IL-12和TNF-α含量显著高于模型对照组（P＜0.05），而IL-10含量显著低于模型对照组（P＜0.05）。综上，超声-闪式联合提取法是一种高效的花色苷提取方式，所得的花色苷提取物可通过诱导细胞凋亡和增强机体免疫抑制肿瘤生长。

关键词: 超声-闪式联合提取, 蓝莓, 花色苷, 工艺, 抗肿瘤活性

Abstract: Ultrasonic-assisted flash extraction (UFE) was employed to extract anthocyanins from blueberry fruit. The effects of ultrasonic power, flash extraction speed, extraction time, solid-to-liquid ratio and ethanol concentration on the yield of anthocyanins were investigated. A combination of one-factor-at-a-time method and genetic algorithm was used to optimize the extraction parameters. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and Annexin V-FITC/PI double label staining were employed to investigate the effects of the anthocyanin extract at different concentrations on proliferation and apoptosis of S180 cells. Its in vivo antitumor activity was tested using an S180 tumor-bearing mice model and enzyme-linked immunosorbent assay (ELISA). The results showed that the optimum extraction conditions that provided the highest yield of anthocyanins of (2.71 ± 0.08) mg/g were obtained as follows: extraction time 10 min, ultrasonic power 450 W, flash extraction speed 8 400 r/min, solid-to-liquid ratio 1:30 (g/mL) and ethanol concentration 68%. The content of anthocyanins in the resulting extract was (31.79 ± 0.53)%. In vitro tests showed that the anthocyanin extract could remarkably inhibit proliferation, induce apoptosis of S180 cells, and block the cells in S phase. In vivo anti-tumor tests showed that the masses of tumors treated with the anthocyanin extract at low, medium and high doses were significantly lower than that of the model control group (P < 0.05) and the anti-tumor rates were 26.36%, 33.33% and 46.51%, respectively. The levels of IL-12 and TNF-α in the serum of tumor-bearing mice treated with the anthocyanins at the three doses were significantly higher than those of the model control group (P < 0.05), while the serum level of IL-10 was significantly lower than that of the model control group (P < 0.05). In conclusion, UFE is an efficient method for anthocyanins extraction, and the anthocyanin extract can inhibit the growth of tumors by inducing apoptosis and enhancing immunity.

Key words: ultrasonic-assisted flash extraction, blueberry, anthocyanins, process, anti-tumor activity

中图分类号:

TS255.42

薛宏坤, 谭佳琪, 刘成海, 刘钗. 超声-闪式联合法制备蓝莓花色苷提取物及其体内外抗肿瘤活性评价[J]. 食品科学, 2020, 41(6): 259-269.

XUE Hongkun, TAN Jiaqi, LIU Chenghai, LIU Chai. Preparation of Blueberry Anthocyanin Extract by Ultrasound-Assisted Flash Extraction and Evaluation of Its Anti-Tumor Activity in Vivo and in Vitro[J]. FOOD SCIENCE, 2020, 41(6): 259-269.

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超声-闪式联合法制备蓝莓花色苷提取物及其体内外抗肿瘤活性评价

Preparation of Blueberry Anthocyanin Extract by Ultrasound-Assisted Flash Extraction and Evaluation of Its Anti-Tumor Activity in Vivo and in Vitro

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