水蜡树果实总黄酮抗脂质过氧化作用

史晓晶; 蔡瑾; 靳辉; 李丹; 张悦悦; 郭军; 吴婷

doi:10.19303/j.issn.1008-0384.2020.12.013

水蜡树果实总黄酮抗脂质过氧化作用

doi: 10.19303/j.issn.1008-0384.2020.12.013

史晓晶^1, ,,
蔡瑾²,
靳辉¹,
李丹¹,
张悦悦¹,
郭军¹,
吴婷¹

1.
忻州师范学院生物系，山西　忻州　034000
2.
山西大学应用化学研究所，山西　太原　030006

基金项目: 山西省高等学校科技创新项目（2019L0832）；山西省应用基础研究青年基金面上项目（201701D221179）

详细信息

通讯作者:
史晓晶（1983−），女，博士，副教授，研究方向：天然活性产物与环境毒理（Email：xzsysxj@sina.com）

中图分类号: S 609+.2
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出版历程
- 收稿日期: 2020-02-06
- 修回日期: 2020-12-31
- 刊出日期: 2020-12-31

Anti-peroxidation of Total Flavonoids from Fruits of Ligustrum obtusifolium on Lipids

1.
Department of Biology, Xinzhou Normal University, Xinzhou, Shanxi　034000, China
2.
Institute of Applied Chemistry, Shanxi University, Taiyuan, Shanxi　030006, China

摘要

摘要: 目的初步探究水蜡树果实总黄酮的抗脂质过氧化作用。方法用分光光度法测定水蜡树果实总黄酮对DPPH·自由基和羟自由基（·OH）的清除率，分析其对小鼠肝匀浆中自氧化和诱导（FeSO₄和H₂O₂）生成的丙二醛（MDA）含量、Fe²⁺与抗坏血酸（Vc）诱导的小鼠肝线粒体肿胀度、H₂O₂诱导的小鼠红细胞氧化溶血、氧化氢酶（CAT）和谷胱甘肽过氧化氢酶（GSH-Px）活性及谷胱甘肽（GSH）含量的影响。结果水蜡树果实总黄酮对DPPH·自由基和·OH自由基具有一定的清除能力，IC₅₀分别为（0.45±0.01） g·L⁻¹和（1.57±0.12 ）g·L⁻¹；1.0～6.0 g·L⁻¹水蜡树果实总黄酮能抑制自氧化体系中MDA的生成；0.5～6.0 g·L⁻¹总黄酮可降低FeSO₄诱导条件下MDA的生成；2.0～6.0 g·L⁻¹总黄酮可抑制H₂O₂诱导条件下MDA的生成；0.5～10.0 g·L⁻¹总黄酮能明显减缓Fe²⁺与Vc诱导的肝线粒体悬液OD₅₂₀值的下降，2.0～10.0 g·L⁻¹总黄酮可明显减弱H₂O₂诱导红细胞的溶血度；0.6～4.0 g·L⁻¹总黄酮可显著提高CAT和GSH-Px的活性及GSH的含量。结论水蜡树果实总黄酮可减弱小鼠肝组织自发性过氧化和诱导脂质过氧化，抑制小鼠肝线粒体和红细胞的氧化损伤，提高抗氧化酶的活性及非酶抗氧化物的含量。本研究对水蜡树果实天然抗氧化剂的开发有一定的借鉴意义。
- 水蜡树果实 /
- 总黄酮 /
- 抗氧化 /
- 脂质过氧化
Abstract: Objective Anti-peroxidation effect of the flavonoids extracted from fruits of Ligustrum obtusifolium on lipids was investigated. Method Scavenging capacities of the crude extract on DPPH· and hydroxyl (·OH) free radicals were tested using spectrophotometry. Effects of the flavonoids on the malondialdehyde (MDA) content in rat liver homogenate upon spontaneous oxidation or peroxidation by either FeSO₄ or H₂O₂ induction and the mitochondrial swelling on rat liver induced by Fe²⁺ and ascorbic acid (Vc) as well as the hemolysis of red blood cells (RBCs) induced by H₂O₂ were assessed in vitro. And the activity of the catalase (CAT) and glutathione peroxidase (GSH-Px) and the level of the glutathione (GSH) were tested in vivo. Result The flavonoid extract exhibited abilities in scavenging DPPH· and ·OH free radicals with IC₅₀ of (0.45±0.01) g·L⁻¹ and (1.57±0.12) g·L⁻¹, respectively. From the concentration of 1.0 g·L⁻¹ to 6.0 g·L⁻¹, the extract significantly reduced the in vitro MDA in rat liver homogenate under spontaneous oxidation; from 0.5 g·L⁻¹ to 6.0 g·L⁻¹, under FeSO₄ induction; and, from 2.0 g·L⁻¹ to 6.0 g·L⁻¹ under H₂O₂ induction. From 0.5 g·L⁻¹ to 10.0 g·L⁻¹, the flavonoids also suppressed the liver mitochondrial swelling induced by Fe²⁺ and Vc; while, from 2.0 g·L⁻¹ to 10.0 g·L⁻¹, the RBC hemolysis induced by H₂O₂, and those from 0.6 g·L⁻¹ to 4.0 g·L⁻¹ could enhance the level of CAT, GSH, and GSH-Px. Conclusion The crude flavonoid extract from the fruits of L. obtusifolium significantly inhibited in vitro on both spontaneous and induced lipid peroxidation reducing the oxidative damage on the mitochondria and RBCs of rat liver, and enhance the acitivity of antioxidant enzymes and the content of non-enzyme antioxidants. The results suggested a promising potential in developing a natural antioxidant from the fruits.
- Ligustrum obtusifolium fruit /
- total flavonoids /
- antioxidation /
- lipid peroxidation

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图 1 水蜡树果实总黄酮粗提物对DPPH·（左）和·OH（右）的清除率

注：同浓度上不同小写字母表示经Duncan氏新复极差法检验在0.05水平下差异显著。

Figure 1. Scavenging capacities of crude flavonoid extract from L. obtusifolium fruit on DPPH· (A) and ·OH (B)

Note: Data with different letters on same concentration indicate significant difference at P＜0.05 by Duncan’s new multiple range test.

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图 2 水蜡树果实总黄酮粗提物对小鼠肝匀浆生成MDA的影响

注：ND代表无诱导组，FD代表FeSO₄诱导组，HD代表H₂O₂诱导组。同色柱上方不同小写字母表示经Duncan氏新复极差法检验在0.05水平下差异显著。

Figure 2. MDA contents in rat liver homogenate treated by flavonoid extract of L. obtusifolium fruits

Note: ND denotes no induction treatment; FD, induction by FeSO₄; and, HD, induction by H₂O₂. Data with different letters on column of same color indicate significant difference at P＜0.05 by Duncan’s new multiple range test.

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图 3 水蜡树果实总黄酮粗提物对小鼠肝线粒体肿胀度的影响

注：FVD代表FeSO₄和Vc共同作用的诱导组，CK为对照。同一时间处理上不同小写字母表示经Duncan氏新复极差法检验在0.05水平下差异显著。

Figure 3. Rat liver mitochondrial swelling induced by Fe²⁺ and Vc treated by flavonoid extract of L. obtusifolium fruits

Note: FVD denotes induction by FeSO₄ and Vc; and, CK, control. Data with different letters on same time treatment indicate significant difference at P＜0.05 by Duncan’s new multiple range test.

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图 4 水蜡树果实总黄酮粗提物对H₂O₂诱导红细胞氧化溶血的影响

注：柱中不同小写字母表示经Duncan氏新复极差法检验在0.05水平下差异显著。

Figure 4. Hemolysis of RBCs treated by flavonoid extract of L. obtusifolium fruits

Note: Different letters in the columns indicate significant difference at P＜0.05 level by Duncan’s new multiple range test.

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表 1 水蜡树果实总黄酮粗提物对小鼠肝组织中CAT、GSH和GSH-Px的影响

Table 1. Effect of crude extract of total flavonoids from L. obtusifolium fruit on CAT, GSH and GSH-Px in liver tissues of rats

组别	CAT/ （U·mgprot^-1）	GSH/ （μmol·gprot^-1）	GSH-Px/ （U·mgprot^-1）
C	20.53±0.47 d	50.78±0.99 d	82.54±0.84 d
L	26.49±0.50 c	60.20±0.67 c	108.40±1.35 c
M	30.84±0.95 b	71.46±1.30 b	123.44±0.90 b
H	34.91±0.61 a	77.47±0.74 a	131.06±0.79 a
注：同列内不同小写字母表示经Duncan氏新复极差法检验在0.05水平下差异显著。 Note: Different letters in the columns indicate significant difference at P＜0.05 level by Duncan’s new multiple range test.

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