红豆越橘果总黄酮保护线粒体及其机制

doi:10.7506/spkx1002-6630-201319015

食品科学

红豆越橘果总黄酮保护线粒体及其机制

李兴泰1，冮洁1，匡海学2，金凤新3，海华1，张雅奎3，刘德文3

1.大连民族学院生命科学学院，辽宁大连 116600；2.黑龙江中医药大学药学院，黑龙江哈尔滨 150040；
3.大兴安岭农林科学院，黑龙江加格达奇 165000

出版日期:2013-10-15 发布日期:2013-09-27
通讯作者: 李兴泰
基金资助:
中央高校基本科研业务费专项(DC12010210)；黑龙江中医药大学博士后科研流动站大兴安岭北奇神绿色产业集团
博士后科研工作站资助项目(LRB10-316)；大连民族学院人才引进科研项目(20116126)

Mitochondrial Protection Activity of Total Flavonoids from Vaccinium vitis-idaea Fruits and Underlying Mechanism

LI Xing-tai1，GANG Jie1，KUANG Hai-xue2，JIN Feng-xin3，HAI Hua1，ZHANG Ya-kui3，LIU De-wen3

1. College of Life Science, Dalian Nationalities University, Dalian 116600, China；
2. College of Pharmacy, Heilongjiang University of Chinese Medicine, Harbin 150040, China；
3. Daxing’anling Academy of Agriculture and Forestry Sciences, Jiagedaqi 165000, China

Online:2013-10-15 Published:2013-09-27
Contact: LI Xing-tai

摘要/Abstract

摘要：

目的：探索红豆越橘果总黄酮(VTF)保护线粒体活性及其机制。方法：用60%乙醇连续回流提取冷冻干燥的红豆越橘果，经大孔吸附树脂纯化后得VTF，并用AlCl3比色法测定其总黄酮含量。以Fe2+/VC诱发肝及脑线粒体脂质过氧化，采用硫代巴比妥酸显色法测定丙二醛(MDA)含量；用Ca2+诱导肝线粒体通透性转换(MPT)，用分光光度法测MPT程度；以还原型辅酶Ⅰ/吩嗪硫酸甲酯(NADH/PMS)为超氧阴离子自由基(O2－•)生成系统，过氧化氢(H2O2)/Fe2+体系为羟自由基(•OH)生成系统，分别用氮蓝四唑(NBT)还原法和Fenton反应显色法测定VTF清除自由基O2－•及•OH的能力；用Na2S2O3滴定法测定VTF清除H2O2的能力；还测定VTF对Fe2+螯合能力及还原力的影响。结果：冷冻干燥的红豆越橘果总黄酮提取率达5.1%，VTF提取物中黄酮的含量为72.3%。VTF可明显抑制线粒体MDA生成；VTF能明显清除O2－•、•OH和H2O2，并均呈剂量-效应关系；VTF具有一定的Fe2+螯合能力及还原力。另外，Ca2+引起的MPT可通过加入VTF而在一定程度上抑制其发生。结论：VTF能通过抗氧化、清除活性氧及抑制MPT来保护线粒体免受损伤，这是VTF保护线粒体的机制。

关键词: 红豆越橘, 总黄酮, 线粒体保护, 活性氧, 线粒体通透性转换

Abstract:

Objective: To investigate the mitochondrial protection activity and underlying mechanism of total flavonoids from Vaccinium vitis-idaea (VTF). Methods: Freeze-dried V. vitis-idaea fruits were extracted by continuous reflux with 60% ethanol followed by purification with macroporous resin to prepare VTF. Total flavonoids were determined by AlCl3 colorimetry. Lipid peroxidation of liver and brain mitochondria was induced by Fe2+/VC in vitro. Thiobarbituric acid
(TBA) colorimetry was used to measure malondialdehyde (MDA) content. Mitochondrial permeability transition (MPT) of rat liver was induced by Ca2+ overload in vitro and measured by spectrophotometric method. The scavenging activities of VTF against superoxide anion (O2 －•) and hydroxyl (•OH) free radicals, which were produced by reduced nicotinamideadenine dinucleotide (NADH)/N-methylphenazonium methyl sulfate (PMS) and hydrogen peroxide (H2O2)/Fe2+ system, respectively, measured by NBT reduction and Fenton reaction colorimetry, respectively. The Na2S2O3 titration method was used to measure the scavenging activities of VTF against H2O2. The effect of VTF on Fe2+ chelation and reducing power was also examined. Results: The extraction yield of flavonoids from freeze-dried V. vitis-idaea fruits was 5.1%, and the content of VTF was 72.3%. VTF could inhibit mitochondrial MDA production and scavenge O2 －•, •OH and H2O2 significantly in a dose-dependent manner, respectively. It could increase Fe2+ chelation and reducing power in some degree and protect mitochondria from permeability transition. Conclusion: The protective effect of VTF on mitochondrial injury is ascribed to antioxidation, scavenging reactive oxygen species (ROS) and inhibiting MPT, which may be the underlying mechanism of mitochondrial protection by VTF.

Key words: Vaccinium vitis-idaea, total flavonoids, mitochondrial protection, reactive oxygen species, mitochondrial permeability transition

中图分类号:

Q731

李兴泰1，冮洁1，匡海学2，金凤新3，海华1，张雅奎3，刘德文3. 红豆越橘果总黄酮保护线粒体及其机制[J]. 食品科学, doi: 10.7506/spkx1002-6630-201319015.

LI Xing-tai1，GANG Jie1，KUANG Hai-xue2，JIN Feng-xin3，HAI Hua1，ZHANG Ya-kui3，LIU De-wen3. Mitochondrial Protection Activity of Total Flavonoids from Vaccinium vitis-idaea Fruits and Underlying Mechanism[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201319015.

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红豆越橘果总黄酮保护线粒体及其机制

Mitochondrial Protection Activity of Total Flavonoids from Vaccinium vitis-idaea Fruits and Underlying Mechanism

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