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中国精品科技期刊2020

柠檬皮多酚成分分析及其对胰岛素抵抗HepG2细胞糖代谢的影响

王瑞雪 张筠 崔艳伟 付红岩 房一明 张彦军 初众

王瑞雪,张筠,崔艳伟,等. 柠檬皮多酚成分分析及其对胰岛素抵抗HepG2细胞糖代谢的影响[J]. 食品工业科技,2022,43(23):310−317. doi:  10.13386/j.issn1002-0306.2022030281
引用本文: 王瑞雪,张筠,崔艳伟,等. 柠檬皮多酚成分分析及其对胰岛素抵抗HepG2细胞糖代谢的影响[J]. 食品工业科技,2022,43(23):310−317. doi:  10.13386/j.issn1002-0306.2022030281
WANG Ruixue, ZHANG Yun, CUI Yanwei, et al. Analysis of Polyphenols from Lemon Peel and Its Effect on Glucose Metabolism in Insulin-resistant HepG2 Cells[J]. Science and Technology of Food Industry, 2022, 43(23): 310−317. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022030281
Citation: WANG Ruixue, ZHANG Yun, CUI Yanwei, et al. Analysis of Polyphenols from Lemon Peel and Its Effect on Glucose Metabolism in Insulin-resistant HepG2 Cells[J]. Science and Technology of Food Industry, 2022, 43(23): 310−317. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022030281

柠檬皮多酚成分分析及其对胰岛素抵抗HepG2细胞糖代谢的影响

doi: 10.13386/j.issn1002-0306.2022030281
基金项目: 农业农村部香辛饮料作物遗传资源利用重点实验室、海南省香料饮料作物工程技术研究中心开放课题资助(2019xys003);黑龙江东方学院科研创新团队建设项目(HDFKYTD202104)。
详细信息
    作者简介:

    王瑞雪(1998−),女,硕士研究生,研究方向:食品功能性,E-mail:1186615232@qq.com

    通讯作者:

    张筠(1976−),女,博士,教授,研究方向:食品功能性,E-mail:forever19@sina.com

    初众(1981−),男,硕士,研究员,研究方向:食品工程,E-mail:cz809@163.com

  • 中图分类号: TQ283

Analysis of Polyphenols from Lemon Peel and Its Effect on Glucose Metabolism in Insulin-resistant HepG2 Cells

  • 摘要: 目的:探究柠檬皮多酚(Limon Peel Polyphenols,LPP)的组成成分,并研究其对胰岛素抵抗(Insulin Resistance,IR)的HepG2细胞糖代谢的影响。方法:采用高效液相色谱串联四级杆飞行时间质谱法(HPLC-QTOF-MS)分析LPP组成,利用HepG2细胞建立胰岛素抵抗模型,用LPP作用于IR-HepG2细胞,通过测定细胞葡萄糖消耗量初步探究LPP对糖代谢的影响,再通过测定糖原含量和己糖激酶(Hexokinase,HK)、丙酮酸激酶(Pyruvate Kinase,PK)、磷酸烯醇式丙酮酸羧基酶(Phosphoenol Pyruvate Carboxykinase,PEPCK)、葡萄糖六磷酸酶(Glucose-6-Phosphatase,G6Pase)的活性,探究LPP调节细胞糖代谢的作用途径。结果:经过HPLC-QTOF-MS分析出12种物质,主要为黄酮及其苷类成分;在糖代谢研究方面,与模型组相比浓度为0.1~2 mg/mL柠檬皮多酚可显著提高细胞葡萄糖消耗量(P<0.05),且当浓度为0.5 mg/mL时其提高IR-HepG2细胞糖原含量和HK、PK活性,降低PEPCK和G6Pase活性的能力最接近于二甲双胍阳性对照。结论:柠檬皮多酚可以缓解IR-HepG2细胞的胰岛素抵抗状态,并能通过促进糖原合成、提高糖酵解关键酶活性、降低糖异生酶活力的方式调节糖代谢水平,为后续的体内研究提供了数据支持,并为未来开发功能性产品提供了理论依据。
  • 图  1  柠檬皮多酚总离子流色谱图

    Figure  1.  TIC chromatography of lemon peel polyphenols

    图  2  柠檬皮多酚高效液相色谱图

    Figure  2.  HPLC chromatography of lemon peel polyphenols

    图  3  峰9(b)的EIC图(A)、一级质谱图(B)和二级质谱图(C)

    Figure  3.  EIC spectra (A)、MS spectra (B) and MS/MS spectra (C) of peak 9(b)

    图  4  LPP对IR-HepG2细胞葡萄糖消耗量的影响

    Figure  4.  Effect of LPP on the glucose consumption of IR-HepG2 cells

    注:相同培养时间条件下,#表示与模型组相比差异显著(P<0.05);△表示与阳性对照组相比差异显著(P<0.05),未标注表示差异不显著(P>0.05)。

    图  5  LPP对IR-HepG2细胞存活率的影响

    Figure  5.  Effect of LPP on survival rate of IR-HepG2 cells

    图  6  LPP对IR-HepG2细胞中糖原含量的影响

    Figure  6.  Effect of LPP on the glycogen content in IR-HepG2 cells

    注:与空白对照组相比,*表示P<0.05,**表示P<0.01,***表示P<0.001;与模型组相比,###表示P<0.001;与阳性对照组相比,△△表示P<0.01,△△△表示P<0.001;未标记表示P>0.05,图7~图10同。

    图  7  LPP对IR-HepG2细胞中HK活性的影响

    Figure  7.  Effect of LPP on the activity of HK in IR-HepG2 cells

    图  8  LPP对IR-HepG2细胞中PK活性的影响

    Figure  8.  Effect of LPP on the activity of PK in IR-HepG2 cells

    图  9  LPP对IR-HepG2细胞中PEPCK活性的影响

    Figure  9.  Effect of LPP on the activity of PEPCK in IR-HepG2 cells

    图  10  LPP对IR-HepG2细胞中G6Pase活性的影响

    Figure  10.  Effect of LPP on the activity of G6Pase in IR-HepG2 cells

    表  1  LPP成分分析结果

    Table  1.   The results of LPP component analysis

    序号保留时间(min)母离子子离子分子式误差名称参考文献
    13.34169.049593 [M+H-甲氧基-CO-羟基]+C8H8O40.06香草酸[18]
    24.93355.1024193 [M+H-葡萄糖]+C16H18O9−0.12新绿原酸[19]
    37.70625.1760487 [M+H-138]+C28H32O160.56Chrysoeriol-7-O-(2'-O-mannopyranosyl)allopyranoside
    47.93449.1080329 [M+H-120]+C21H20O11−0.21荭草素[20]
    59.18433.1131283 [M+H-阿拉伯糖]+C21H20O10−0.85山奈黄苷[21]
    69.19597.1815289 [M+H-新橙皮糖]+C27H32O15−0.27圣草枸橼苷[22]
    79.37579.1708271 [M+H-新橙皮糖]+
    433 [M+H-鼠李糖]+
    C27H30O14−0.56野漆树苷[17]
    810.26463.1233313 [M+H-阿拉伯糖]+C22H22O11−0.4金雀花素
    9(a)11.94609.1813301 [M+H-新橙皮糖]+
    463 [M+H-鼠李糖]+
    C28H32O15−0.86地奥司明[23]
    9(b)11.98611.1960303 [M+H-鼠李糖-葡萄糖]+
    465 [M+H-鼠李糖]+
    C28H34O150.39橙皮苷[17]
    1012.41353.0855147 [M+H-206]+C16H16O90.614-甲基-7-乙酰氧基香豆素-β-D-葡萄糖醛酸苷
    1124.59515.2277161 [M+H-C18H28O8]+
    411 [M+H-C3H4O4]+
    C28H34O9−0.2诺米林[24]
    下载: 导出CSV
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  • 收稿日期:  2022-03-23
  • 网络出版日期:  2022-10-21
  • 刊出日期:  2022-11-23

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