纳米硒对低氧胁迫下中华绒螯蟹免疫保护和抗氧化能力的影响

贾慧凝; 侍苗苗; 卞永乐; 侍崇敬; 刘恒蔚; 宋学宏; 秦粉菊

doi:10.12131/20220106

纳米硒对低氧胁迫下中华绒螯蟹免疫保护和抗氧化能力的影响

doi: 10.12131/20220106

1.
苏州科技大学化学与生命科学学院，江苏苏州 215009
2.
苏州大学基础医学与生物科学学院，江苏苏州 215123

基金项目: 国家自然科学基金项目 (31772896)；苏州市科技计划项目 (SNG2017055)

详细信息

作者简介:
贾慧凝 (1998—)，女，硕士研究生，研究方向为环境生物技术。E-mail: jiahuining_a@163.com

通讯作者:
秦粉菊 (1976—)，女，教授，博士，研究方向为环境毒理学和纳米营养学。E-mail: qinfenju@mail.usts.edu.cn

中图分类号: S 963.73⁺4
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出版历程
- 收稿日期: 2022-04-19
- 修回日期: 2022-06-06
- 录用日期: 2022-06-14
- 网络出版日期: 2022-06-25
- 刊出日期: 2022-12-05

Effects of nanometer selenium on immune protection and antioxidant ability of Eriocheir sinensis under hypoxia stress

1.
School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou 215009, China
2.
School of Biology and Basic Medical Sciences, Soochow University, Suzhou 215123, China

摘要

摘要: 低氧胁迫会减弱中华绒螯蟹 (Eriocheir sinensis) 的免疫机能和抗氧化能力。为揭示纳米硒 (nano-Se) 对低氧胁迫下中华绒螯蟹的免疫保护作用及抗氧化调控机制，在基础饲料中添加不同水平 (0、0.1、0.2、0.4、0.8和1.6 mg·kg⁻¹) 的纳米硒饲喂中华绒螯蟹90 d。饲喂实验结束后，进行低氧胁迫实验并注射嗜水气单胞菌 (Aeromonas hydrophila)。结果表明：1) 低氧胁迫24 h和低氧胁迫下感染嗜水气单胞菌24 h的蟹死亡率分别可达62.45%和100%，低氧胁迫12 h使血淋巴中血蓝蛋白浓度、血细胞数量、组织中超氧化物歧化酶 (SOD)、过氧化氢酶 (CAT)、谷胱甘肽过氧化物酶 (GSH-Px) 活性显著升高 (P<0.05)，胁迫至第24小时有下降趋势；乳酸 (LD) 和丙二醛 (MDA) 浓度在低氧胁迫12~24 h持续上升。2) 饲料中添加适量 (0.1~0.4 mg·kg⁻¹) 纳米硒可显著降低低氧胁迫下蟹死亡率和低氧胁迫下嗜水气单胞菌的致死率 (P<0.05)，显著提高低氧胁迫下血蓝蛋白浓度和血细胞数量以及抗氧化酶 (SOD、CAT、GSH-Px) 活性，降低LD和MDA浓度 (P<0.05)；添加0.8~1.6 mg·kg⁻¹纳米硒加剧了低氧胁迫损伤。结果表明饲料中添加适量纳米硒可改善低氧胁迫下中华绒螯蟹的免疫功能和抗氧化能力，且添加水平以0.2 mg·kg⁻¹为宜。
- 中华绒螯蟹 /
- 纳米硒 /
- 低氧胁迫 /
- 免疫 /
- 抗氧化
Abstract: Hypoxia stress will weaken the immune function and antioxidant capacity of Eriocheir sinensis. In order to reveal the immune protection and anti-oxidation regulation mechanism of nanometer selenium (nano-Se) on E. sinensis under hypoxia stress, we had fed E. sinensis with different doses of nano-Se (0, 0.1, 0.2, 0.4, 0.8 and 1.6 mg·kg⁻¹) in basic diets for 90 d. After the feeding, we conducted a hypoxia stress test and injected Aeromonas hydrophila under hypoxia stress. The results show that: 1) The mortality of E. sinensis under hypoxia stress for 24 h and that infected with A. hydrophila under hypoxia stress reached 62.45% and 100%, respectively. The levels of hemocyanin and the hemocyte count in crab hemolymph, and the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in crab tissues increased significantly under hypoxia stress for 12 h (P<0.05), as well as there was a decreasing trend under stress to 24 h. The contents of lactic acid (LD) and malondialdehyde (MDA) continued to rise from 12 to 24 h under hypoxia stress. 2) Appropriate addition amount (0.1−0.4 mg·kg⁻¹) of nano-Se reduced the mortality of E. sinensis significantly and the lethality of A. hydrophila under hypoxia stress (P<0.05), increasing the levels of hemocyanin, the hemocyte count and the activities of antioxidant enzymes (SOD, CAT, GSH-Px) under hypoxia stress significantly, but decreasing the contents of LD and MDA (P<0.05). The addition of 0.8−1.6 mg·kg⁻¹ nano-Se had aggravated hypoxia stress injury. These results indicate that appropriate addition of nano-Se to the diets can improve the decrease of immune response and antioxidant ability of E. sinensis under hypoxia stress, and the optimal dose of nano-Se in basal diets is 0.2 mg·kg⁻¹.
- Eriocheir sinensis /
- Nanometer selenium /
- Hypoxia stress /
- Immunity /
- Antioxidant

HTML全文

图 1 纳米硒对低氧胁迫下中华绒螯蟹血淋巴耐低氧指标的影响 (N=6)

注：柱上不同英文字母表示相同低氧胁迫时间下显著差异 (P<0.05)；*. 与对照组差异显著 (P<0.05)；后图同此。

Figure 1. Effects of nano-Se on hemolymph hypoxia tolerance indexes of E. sinensis under hypoxia stress (N=6)

Note: Different lowercase letters on the bar indicate significant difference at the same hypoxia stress time (P<0.05); *. Significant difference compared with the control group (P<0.05); the same case in the following figures.

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图 2 纳米硒对低氧胁迫下中华绒螯蟹血淋巴血细胞数量的影响 (N=6)

Figure 2. Effects of nano-Se on hemolymph hemocyte count of E. sinensis under hypoxia stress (N=6)

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图 3 纳米硒对低氧胁迫下中华绒螯蟹不同组织中超氧化物歧化酶活性的影响 (N=6)

Figure 3. Effects of nano-Se on SOD activity in different tissues of E. sinensis under hypoxia stress (N=6)

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图 4 纳米硒对低氧胁迫下中华绒螯蟹不同组织中过氧化氢酶活性的影响 (N=6)

Figure 4. Effects of nano-Se on CAT activity in different tissues of E. sinensis under hypoxia stress (N=6)

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图 5 纳米硒对低氧胁迫下中华绒螯蟹不同组织中谷胱甘肽过氧化物酶活性的影响 (N=6)

Figure 5. Effects of nano-Se on GSH-Px activity in different tissues of E. sinensis under hypoxia stress (N=6)

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图 6 纳米硒对低氧胁迫下中华绒螯蟹不同组织中丙二醛浓度的影响 (N=6)

Figure 6. Effects of nano-Se on MDA concentration in different tissues of E. sinensis under hypoxia stress (N=6)

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表 1 基础饲料配方组成

Table 1. Ingredients of composition of basal diets

原料 Ingredient	质量分数 Mass fraction/%
鱼粉 Fish meal	17
棉粕 Cotton seed meal	17
菜粕 Rape seed meal	16
次粉 Wheat flour	10.5
豆粕 Soybean meal	10
玉米 Corn	9
米糠 Rice bran	5
黏合剂 Adhesive	1
血粉 Blood meal	3
虾壳粉 Shrimp shell meal	3
豆油+菜油 (1∶1) Soybean oil+ Rapeseed oil	2
磷酸二氢钙 Ca(H₂PO₄)₂	1.5
沸石粉 Zeolite powder	2
河蟹饲料添加剂 Crab feed additive	1
蟹用多维^① Crab vit premix	1
蟹用多矿^② Crab min premix	1
注：①. 每100 g蟹用多维预混料中含：维生素E 2.0 g、维生素C 3.0 g、维生素A 0.6 g、维生素D₃ 0.08 g、维生素B₁ 0.07 g、维生素B₂ 0.14 g、维生素B₃ 0.28 g、维生素B₅ 0.01 g、维生素B₆ 0.08 g、维生素B₇ 0.05 g、维生素B₁₁ 0.02 g、维生素H 0.04 g、烟酸0.3 g、叶酸0.05 g、氯化胆碱0.5 g、泛酸钙0.25 g、生物素0.05 g、肌醇0.7 g；②. 每100 g蟹用多矿预混料中含：磷酸二氢钠3.5 g、磷酸二氢钾6.0 g、碳酸钙3.5 g、氯化钾0.6 g、七水合硫酸镁3.2 g、六水合氯化铝0.55 g、七水合硫酸锌0.157 g、柠檬酸铁0.019 g、四水合硫酸锰0.043 g、碘化钾0.016 g、氯化铜0.014 g、六水合氯化钴0.055 g、乳酸钙5.15 g。 Note: ①. Per 100 g of crab multi vitamin premix contains: ${\rm{V}}_{\rm{E}} $ 2.0 g , ${\rm{V}}_{\rm{C}} $ 3.0 g, ${\rm{V}}_{\rm{A}} $ 0.6 g, ${\rm{V}}_{{\rm{D}}_3}$ 0.08 g, ${\rm{V}}_{{\rm{B}}_1}$ 0.07 g, ${\rm{V}}_{{\rm{B}}_2}$ 0.14 g, ${\rm{V}}_{{\rm{B}}_3}$ 0.28 g, ${\rm{V}}_{{\rm{B}}_5}$ 0.01 g, ${\rm{V}}_{{\rm{B}}_6}$ 0.08 g, ${\rm{V}}_{{\rm{B}}_7}$ 0.05 g, ${\rm{V}}_{{\rm{B}}_{11}}$ 0.02 g, V_H 0.04 g, niacin 0.3 g, folic acid 0.05 g, choline chloride 0.5 g, calcium pantothenate 0.25 g, biotin 0.05 g, inositol 0.7 g; ②. Per 100 g of crab multi mineral premix contains: NaH₂PO₄ 3.5 g, KH₂PO₄ 6.0 g, CaCO₃ 3.5 g, KCl 0.6 g, MgSO₄·7H₂O 3.2 g, AlCl₃·6H₂O 0.55 g, ZnSO₄·7H₂O 0.157 g, FeC₆H₅O₇ 0.019 g, MnSO₄·4H₂O 0.043 g, KI 0.016 g, CuCl₂ 0.014 g, CoCl₂·6H₂O 0.055 g, C₆H₁₀CaO₆ 5.15 g.

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表 2 纳米硒对低氧胁迫下中华绒螯蟹死亡率的影响 (N=10)

Table 2. Effects of nano-Se on mortality rate of E. sinensis under hypoxia stress (N=10)

纳米硒添加水平 Level of nano-Se/(mg·kg⁻¹)	死亡率 Mortality rate/%			免疫保护率 Immune protection rate/%
纳米硒添加水平 Level of nano-Se/(mg·kg⁻¹)	0 h	12 h	24 h	12 h	24 h
0	0	12.66±3.33^c	62.45±8.57^c*	—	—
0.1	0	6.25±1.50^b	44.39±4.88^b	50.63	28.92
0.2	0	2.50±0.30^a	25.62±3.33^a	80.25	58.98
0.4	0	12.34±2.05^c	31.55±3.29^b	2.53	49.48
0.8	0	19.36±2.67^d	62.66±6.32^c	−52.92	−0.34
1.6	0	20.25±5.55^d	87.73±10.34^d	−59.95	−40.48
注：同列不同上标字母表示差异显著 (P<0.05)；. 与对照组 (0 mg·kg⁻¹纳米硒低氧胁迫0 h组) 差异显著 (P<0.05)；—. 无数据；后表同此。 Note: Different superscript letters within the same column indicate significant difference (P<0.05); . Significant difference compared with the control group (0 mg·kg⁻¹ nano-Se group under hypoxia stress for 0 h) (P<0.05); —. No data. The same case in the following tables.

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表 3 纳米硒对低氧胁迫下嗜水气单胞菌致死率的影响 (N=10)

Table 3. Effects of nano-Se on lethality of A. hydrophila under hypoxia stress (N=10)

纳米硒添加水平 Level of nano-Se/(mg·kg⁻¹)	嗜水气单胞菌致死率 Lethality of A. hydrophila/%			免疫保护率 Immune protection rate/%
纳米硒添加水平 Level of nano-Se/(mg·kg⁻¹)	0 h	12 h	24 h	12 h	24 h
0	0	50.36±6.72^c*	100±0^c*	—	—
0.1	0	25.22±3.51^b	87.47±5.95^b	49.92	12.53
0.2	0	12.48±2.39^a	56.82±7.31^a	75.22	43.18
0.4	0	25.78±3.64^b	81.43±2.5^b	48.81	18.57
0.8	0	50.33±4.52^c	100±0^c	0.06	0
1.6	0	75.29±6.15^d	100±0^c	−49.5	0

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