半番鸭不同组织Cu/Zn-SOD基因表达及酶活性研究

郑嫩珠; 章琳俐; 李丽; 辛清武; 缪中纬; 朱志明; 黄一帆

doi:10.19303/j.issn.1008-0384.2019.08.010

半番鸭不同组织Cu/Zn-SOD基因表达及酶活性研究

doi: 10.19303/j.issn.1008-0384.2019.08.010

郑嫩珠^{1, 2,},
章琳俐²,
李丽²,
辛清武²,
缪中纬²,
朱志明²,
黄一帆^1, ,

1.
福建农林大学食品科学学院, 福建福州 350002
2.
福建省农业科学院畜牧兽医研究所, 福建福州 350013

基金项目:

中央引导地方科技发展专项 2018L3004

福建省发改委"五新"项目 016032018001

福建省财政专项——福建省农业科学院创新团队建设项目 STIT2017-2-1

详细信息

作者简介:
郑嫩珠(1972-), 女, 博士, 研究员, 研究方向:畜禽遗传育种与农产品加工(E-mail:hql202@126.com)

通讯作者:
黄一帆(1954-), 男, 教授, 博士生导师, 研究方向:农产品加工与贮藏(E-mail:zjhyfang@163.com)

中图分类号: X131;X53
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出版历程
- 收稿日期: 2019-07-10
- 修回日期: 2019-08-03
- 刊出日期: 2019-08-01

Expression of Cu/Zn-SOD Gene and Activity of Cu/Zn-SOD Enzyme in Mule Duck Tissues

1.
College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
2.
Institute of Animal Science and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China

摘要

摘要: 目的首次对半番鸭不同组织Cu/Zn-SOD基因表达、酶活性进行研究，以期了解半番鸭不同组织抗氧化特性，为深入研究半番鸭对环境因子的响应能力及相关分子机制提供参考。方法通过实时荧光定量法、黄嘌呤氧化酶法和硫代巴比妥酸法分别测定了半番鸭心脏、肝脏、脾脏、肺脏、肾脏、肌胃和胸肌中的Cu/Zn-SOD mRNA相对表达量、酶活力和MDA含量。结果 Cu/Zn-SOD mRNA在胸肌中相对表达量最高，总体趋势为胸肌>肝脏>肺>心脏>脾脏>肾脏/肌胃。酶活力测定结果显示，肝脏和肾脏Cu/Zn-SOD酶活力显著高于胸肌和肌胃。半番鸭胸肌MDA含量显著高于肝脏、肌胃和肾脏，提示胸肌可能更易受到氧化应激损伤。相关分析结果显示Cu/Zn-SOD活力与MDA含量呈显著负相关，而抗氧化酶活力与mRNA表达水平相关性不显著。结论半番鸭Cu/Zn-SOD基因表达和酶活力具有组织特异性。不同组织Cu/Zn-SOD活力差异可能与Cu/Zn-SOD自身的生理功能以及组织代谢水平有关。
- Cu/Zn-SOD /
- mRNA表达 /
- 酶活力 /
- 半番鸭
Abstract: Objective Expressions of Cu/Zn-SOD gene and activities of the antioxidant enzyme in various organs of mule ducks were determined. Method The gene expression in the heart, liver, spleen, lung, kidney, gizzard and chest muscles, antioxidant enzyme activities, and MDA contents of mule duck were determined by the qRT-PCR, xanthine-xanthine oxidase, and thiobarbituric acid methods. Result The greatest mRNA expression of Cu/Zn-SOD gene was found in the chest muscle followed by the tissues in the order of liver > lung > heart > spleen > gizzard/kidney of the ducks. The activities of Cu/Zn-SOD enzyme in the liver and kidney were significantly higher than those in the breasts and gizzard. The higher MDA content in the breasts than in the liver, gizzard or kidney indicated a greater susceptibility of the chest muscle to damage by oxidative stress. There was no significant correlation observed between the antioxidant enzyme activity and the mRNA expression level in different organs. However, the Cu/Zn-SOD enzyme activity was inversely correlated to MDA content in the tissues. Conclusion The Cu/Zn-SOD mRNA expression and enzyme activity were organ-specific. The differences in the enzymatic activities might stem from the diverse physiological functions of the enzymes and/or the tissue metabolism per se.
- Cu/Zn-SOD /
- mRNA expression /
- enzyme activity /
- mule duck

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图 1 目的基因qRT-PCR扩增曲线和熔解曲线

注：A为SOD1扩增曲线(左)和熔解曲线(右)，B为β-actin基因扩增曲线(左)和熔解曲线(右)。

Figure 1. Amplification and melting curves of qRT-PCR on target genes

Note:A: SOD1 gene amplification curve (left) and melting curve (right); B: β-actin amplification curve (left) and melting curve (right).

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图 2 半番鸭不同组织Cu/Zn-SOD mRNA的相对表达

注：1：心脏；2：肝脏；3：脾脏；4：肺脏；5：胸肌；6：肌胃；7：肾脏。不同小写字母表示在P＜0.05水平差异有统计学意义，不同大写字母表示在P＜0.01水平差异有高度统计学意义。

Figure 2. Relative expressions of Cu/Zn-SOD mRNA in mule duck tissues

Note: 1: heart; 2: liver; 3: spleen; 4: lung; 5: chest; 6: gizzard; and, 7: kidney. Data with different lowercase letters indicate significant differences at P < 0.05; and, data with different capital letters indicate significant differences at P < 0.01.

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图 3 抗氧化酶活力与mRNA、MDA含量相关性分析

Figure 3. Correlation coefficients between antioxidant enzyme activity and mRNA expression/MDA content

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表 1 不同时期饲料的营养水平

Table 1. Nutrients in diets for mule ducks at various stages

营养水平 Nutrient levels/%	0~21 日龄 0~21 days of age	22~42 日龄 22~42 days of age	43~70 日龄 43~70 days of age
粗蛋白 CP	≥18.0	≥16.0	≥15.0
粗纤维 CF	≤5.0	≤6.0	≤7.0
粗灰分 CA	≤8.0	≤12.0	≤12.0
钙 Ca	0.70~1.40	0.70~1.40	0.70~1.40
总磷 AP	≥0.40~0.80	≥0.35~0.75	≥0.35~0.75
食盐 NaCl	0.30~0.80	0.30~0.80	0.30~0.80
水分 Water	≤13.0	≤13.0	≤13.0

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表 2 荧光定量PCR引物信息

Table 2. Information on qRT-PCR primers

基因 Genes	登录号 GenBank No.	引物序列(5′→3′) Primer Sequences (5′to 3′)	扩增长度 Size/bp	Tm /℃
SOD1	KU048808.1	F: CAGCAGGGTAACGGACCAGTAAA；R: CCCAAATTCGTGGACATGGAAG	87	60
ACTB	EF667345	F: GATGTGGATCAGCAAGCAGGAGT；R: GGGTGTGGGTGTTGGTAACAGT	95	60
注：SOD1：铜锌超氧化物歧化酶；ACTB：β-肌动蛋白。 Note: SOD1: Cu/Zn SOD; ACTB: β-actin.

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表 3 半番鸭不同组织Cu/Zn-SOD活力和MDA含量

Table 3. Cu/Zn-SOD enzyme activity and MDA content in mule duck tissues

指标 Index	肝脏 Liver	胸肌 Chest muscle	肌胃 Gizzard	肾脏 Kidney
Cu/Zn-SOD活力Cu/Zn-SOD activity/(U·mg^-1)	176.07±4.57Aa	39.37 ±1.41Cc	48.49±2.08Cc	117.64±4.85 Bb
MDA含量MDA content/(n mol·mg^-1)	1.07±0.02Ba	2.84±0.18 Aa	1.10±0.03 Bb	0.89±0.02 Cc
注：同行数据后不同小写字母表示差异显著(P＜0.05)，不同大写字母表示差异极显著(P＜0.01)。 Note: In the same row, different lowercase letters represent statistically significant differences at the 0.05 probability level; different capital letters represent highly statistically significant differences at the 0.01 probability level.

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