食品科学

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稠李花色苷酶法制备及对H2O2诱导大鼠胰岛素瘤细胞损伤的保护作用

刘 晓,曹向宇,李其久,孙宇航,杨思敏,于 慧,王纬宇,刘剑利*   

  1. 辽宁大学生命科学院,辽宁 沈阳 110036
  • 出版日期:2016-03-25 发布日期:2016-03-18

Enzymatic Preparation of Anthocyanins from Padus racemosa Fruits and Protection against Ins-1 Cell Damage Induced by H2O2

LIU Xiao, CAO Xiangyu, LI Qijiu, SUN Yuhang, YANG Simin, YU Hui, WANG Weiyu, LIU Jianli*   

  1. Liaoning University Academy of Science, Shenyang 110036, China
  • Online:2016-03-25 Published:2016-03-18

摘要:

目的:探究纤维素酶酶法制备稠李花色苷的最佳条件,并研究制备的稠李花色苷对H2O2诱导的大鼠胰岛素瘤(Ins-1)细胞损伤的保护作用。方法:通过单因素试验和正交试验,优化得到酶法制备稠李花色苷的最佳条件;稠李花色苷处理大鼠Ins-1细胞,进行形态学观察、3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]细胞活力实验、2’,7’-二氢二氯荧光素二乙酸酯(2’,7’-dichlorodihydrofluorescein diacetate,DCFH-DA)荧光探针法检测细胞内活性氧实验研究稠李花色苷对H2O2诱导损伤的Ins-1细胞保护作用。结果:纤维素酶法提取稠李花色苷的最佳条件为:温度60 ℃、纤维素酶添加量9 mg/g、料液比1∶35(g/mL)、pH 3.5,此条件下稠李花色苷得率为(0.956±0.027) mg/g;形态学观察及MTT细胞活力实验显示,稠李花色苷对H2O2诱导损伤的Ins-1细胞具有较强的保护作用,DCFH-DA法检测细胞内活性氧实验说明,稠李花色苷能够显著地清除Ins-1细胞内的活性氧。结论:纤维素酶法制备稠李花色苷是一种有效的方法,制备的稠李花色苷对H2O2诱导的大鼠Ins-1细胞损伤具有较强的保护作用。

关键词: 花色苷, 酶法, 氧化损伤, 保护, 大鼠胰岛素瘤(Ins-1)细胞

Abstract:

Purpose: To optimize the extraction conditions for anthocyanins from Padus racemosa fruits by cellulose
hydrolysis, and to detect the protective effect of the extracted anthocyanins on H2O2-induced damage in Ins-1 cells. Methods:
The optimization was performed by single factor and orthogonal array experiments. The protective effect of Padus racemosa
anthocyanins on Ins-1 cells were examined by morphological observation, MTT assay and DCFH-DA fluorescent probe
method. Results: The optimal extraction conditions were achieved by carrying out cellulose hydrolysis at 60 ℃, pH 3.5 and
an enzyme dosage of 9 mg/g with a solid/liquid ratio of 1:35 (g/mL). Under these conditions, the yield of Padus racemosa
anthocyanins was (0.956 ± 0.027) mg/g. Padus racemosa anthocyanins protected Ins-1 cells from damage induced by H2O2
as indicated by morphological observation and MTT assay, and strongly scavenged reactive oxygen species in the DCFHDA
fluorescent assay. Conclusions: Enzymatic hydrolysis is an effective method for extracting anthocyanins from Padus
racemosa, and the extracted anthocyanins strongly protect against H2O2-induced damage in Ins-1 cells.

Key words: anthocyanin, enzyme, oxidative damage, protection, rat insulinoma (Ins-1) cell

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