食品科学 ›› 2022, Vol. 43 ›› Issue (19): 200-207.doi: 10.7506/spkx1002-6630-20210906-067

• 营养卫生 • 上一篇    下一篇

豆芋异黄酮的制备及其对RIN-m5F细胞氧化损伤的保护作用

邓珂,陈镠,杨良缘,胡钰,许光治,张有做,王艳,倪勤学   

  1. (浙江农林大学食品与健康学院,浙江 杭州 311300)
  • 出版日期:2022-10-15 发布日期:2022-10-26
  • 基金资助:
    浙江省自然科学基金项目(LY19C200016)

Preparation of Isoflavones from the Tuber of Apios americana Medik. and Their Protective Effects on RIN-m5F Cells from Oxidative Damage

DENG Ke, CHEN Liu, YANG Liangyuan, HU Yu, XU Guangzhi, ZHANG Youzuo, WANG Yan, NI Qinxue   

  1. (School of Food and Health, Zhejiang Agriculture & Forest University, Hangzhou 311300, China)
  • Online:2022-10-15 Published:2022-10-26

摘要: 以豆芋(Apios americana Medik.)为原料,采用乙醇热回流提取、乙酸乙酯萃取,以异黄酮质量浓度为指标,优化大孔树脂富集工艺,并对得到的豆芋异黄酮(记为AI-3)抗氧化活性、α-葡萄糖苷酶抑制能力及对胰岛瘤细胞RIN-m5F氧化损伤的保护作用进行探究。结果:优化的D101型大孔树脂富集工艺为:上样质量浓度1.5 mg/mL,体积分数80%乙醇溶液(pH 6)洗脱,洗脱体积80 mL(4 倍柱体积),所得AI-3得率为0.44%(占豆芋干基质量),异黄酮质量分数50.83%;AI-3对1,1-二苯基-2-三硝基苯肼自由基、2,2’-联氮-双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基的半抑制质量浓度分别为19.51、3.40 μg/mL,铁离子还原抗氧化能力为387.83 μmol/mg,对α-葡萄糖苷酶活力的半抑制质量浓度为235.97 μg/mL;AI-3在0~300 μg/mL范围内,RIN-m5F细胞存活率同空白对照组相比无显著差异(P>0.05),且25~300 μg/mL时可极显著提高RIN-m5F氧化损伤细胞的存活率(P<0.01),300 μg/mL时细胞存活率达84.19%,高于阳性对照组。结论:D101型大孔树脂可有效富集豆芋异黄酮,所得AI-3具有较强的抗氧化、α-葡萄糖苷酶抑制活性及RIN-m5F细胞氧化损伤保护作用。研究结果为开发预防糖尿病等代谢综合征功能性食品提供了理论依据。

关键词: 豆芋异黄酮;大孔树脂富集;抗氧化;α-葡萄糖苷酶抑制;RIN-m5F细胞保护

Abstract: The intention of this study was to optimized the enrichment process of isoflavones from the tuber of Apios americana Medik. using macroporous adsorption resin and to evaluate their bioactivities. Isoflavones from the tuber of A. americana Medik. were extracted by successive hot ethanol reflux extraction and ethyl acetate extraction, and then the concentration process of isoflavones was optimized based on isoflavone concentration. The antioxidant activity and α-glucosidase inhibitory activity of the obtained isoflavone-rich fraction (AI-3) and its protective effect against oxidative damage to RIN-m5F cells were studied. The optimized processing parameters were as follows: use of D101-type macroporous resin as sorbent, sample concentration of 1.5 mg/mL, use of 80% ethanol as eluent, and eluent volume of 80 mL (the bed volume (BV) was 20 mL). The yield of AI-3 from the dried tuber of A. americana Medik. was 0.44%, and the isoflavone content was 50.83%. The half maximal inhibitory concentration (IC50) of AI-3 for scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation radical were 19.51 and 3.40 μg/mL, respectively, and the ferric reducing capacity was 387.83 μmol/mg. The IC50 against α-glucosidase was 235.97 μg/mL. The survival rate of RIN-m5F cells treated with AI-3 at concentrations in the range of 0–300 μg/mL had no significant difference from that of the control group (P > 0.05), and AI-3 concentrations in the 25–300 μg/mL range significantly improved the survival rate of oxidatively damaged cells (P < 0.01). At 300 μg/mL AI-3 concentration, the survival rate of RIN-m5F cells was 84.19%, which was higher than that of the positive control group. D101-type macroporous resin can effectively enrich isoflavones from the tuber of A. americana Medik. yielding a fraction (AI-3) with strong antioxidant activity, α-glucosidase-inhibitory capacity and protective effect against oxidative damage to RIN-m5F cells. This study will provide a theoretical basis for the development of functional foods for the prevention of diabetes and other metabolic syndromes.

Key words: Apios americana Medik. isoflavones; macroporous resin enrichment; antioxidant; α-glucosidase inhibition; RIN-m5F cell protection

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