亚硝酸盐和微塑料胁迫对凡纳滨对虾鳃中免疫、解毒代谢和渗透调节相关指标的影响

邢逸夫; 段亚飞; 韦政坤; 朱轩仪; 黄建华; 张家松

doi:10.12131/20220176

亚硝酸盐和微塑料胁迫对凡纳滨对虾鳃中免疫、解毒代谢和渗透调节相关指标的影响

doi: 10.12131/20220176

邢逸夫^{1, 2,},
段亚飞²,
韦政坤^{1, 2},
朱轩仪²,
黄建华²,
张家松^2, ,

1.
大连海洋大学水产与生命学院，辽宁大连 116023
2.
中国水产科学研究院南海水产研究所/农业农村部南海渔业资源开发利用重点实验室，广东广州 510300

基金项目: 国家自然科学基金项目 (31902343)；广东省基础与应用基础研究基金 (2021A1515012084)；广州市科技计划项目 (202102080246)；广州市科学技术协会青年人才托举工程项目 (X20210201039)；农业科研杰出人才培养计划 (13210308)；中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助 (2021SD19, 2022RC01)；中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金资助 (2021XT0604)；国家重点研发计划“蓝色粮仓科技创新”专项 (2019YFD0900500)

详细信息

作者简介:
邢逸夫 (1998—)，男，硕士研究生，研究方向为水产动物环境生理。E-mail: xyf907051631@163.com

通讯作者:
张家松 (1971—)，男，研究员，博士，研究方向为设施渔业养殖技术。E-mail: jiasongzhang@hotmail.com

中图分类号: S 917.4
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- 文章访问数: 233
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- 被引次数: 0
出版历程
- 收稿日期: 2022-06-22
- 修回日期: 2022-08-10
- 录用日期: 2022-09-11
- 网络出版日期: 2022-09-16
- 刊出日期: 2023-04-05

Effects of nitrite and microplastic stress on immune, detoxification metabolism and osmoregulation-related indicators in gills of Litopenaeus vannamei

XING Yifu^{1, 2
,},
DUAN Yafei²,
WEI Zhengkun^{1, 2},
ZHU Xuanyi²,
HUANG Jianhua²,
ZHANG Jiasong^{2
, ,}

1.
College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China
2.
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences/Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, Guangzhou 510300, China

摘要

摘要: 鳃是对虾重要的呼吸器官，是亚硝酸盐毒性效应的主要靶器官，也是微塑料主要富集的部位之一。鳃组织参与了对虾渗透调节、氮排泄、免疫功能等生理过程，对对虾维持机体健康具有重要意义。为研究亚硝酸盐和微塑料单因素及复合胁迫对凡纳滨对虾 (Litopenaeus vannamei) 鳃组织生理功能的影响，将对虾分为对照组、20 mg·L⁻¹亚硝酸盐胁迫组 (NIT)、10 µg·L⁻¹微塑料胁迫组 (MP)、20 mg·L⁻¹亚硝酸盐和10 µg·L⁻¹微塑料复合胁迫组 (NM)，于第14天测定对虾鳃中相关指标变化。结果显示：1) 氧化应激指标丙二醛 (MDA)、过氧化氢 (H₂O₂) 浓度及抗氧化酶超氧化物歧化酶 (SOD)、过氧化氢酶 (CAT)、谷胱甘肽过氧化物酶 (GPx) 活性在胁迫后发生不同程度的改变；2) 解毒代谢指标细胞色素P450基因 (CYP450) 和谷胱甘肽S-转移酶基因 (GST) 相对表达水平在胁迫后发生不同程度的紊乱；3) 渗透调节指标如离子转运酶液泡型ATP酶 (VATP)、Na⁺/H⁺交换体7 (NHE7)、Na⁺/K⁺-ATP酶亚基α (NKA-α)、Na⁺/K⁺-ATP酶亚基β (NKA-β)、碳酸酐酶 (CA) 和水通道蛋白TIP4-1 (TIP4)、钙通道蛋白1 (CCP)、氯通道蛋白2 (CLC)、水通道蛋白 (AQP) 基因的相对表达水平在胁迫后发生不同程度的紊乱；4) 细胞凋亡因子基因 (CASP-3) 相对表达水平在3个胁迫组均显著降低 (P<0.05)。由此推断，亚硝酸盐和微塑料胁迫会诱导凡纳滨对虾鳃中免疫、解毒代谢和渗透调节相关指标变化，进而影响其正常的生理功能。
- 凡纳滨对虾 /
- 鳃 /
- 亚硝酸盐 /
- 微塑料 /
- 解毒代谢 /
- 渗透调节
Abstract: Gill is an important respiratory organ of prawns, the main target organ of nitrite toxicity effect, and also one of the main enrichment sites of microplastics. The gill tissue participates in the physiological processes of prawns such as osmoregulation, nitrogen excretion, immune function, etc., which is important for maintaining the prawns' health. In order to investigate the effects of single and combined stress of nitrite and microplastics on the physiological functions of gill tissues of Litopenaeus vannamei, we designed the control group, 20 mg·L⁻¹ nitrite stress group (NIT), 10 μg·L⁻¹ microplastic stress group (MP), 20 mg·L⁻¹ nitrite and 10 μg·L⁻¹ microplastic composite stress group (NM), and then measured the changes of immune and osmotic regulation in gills of shrimps on the 14^th day. The results show that: 1) The oxidative stress indicators such as the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) changed at different degrees after the stress. 2) The detoxification metabolic indicators such as the relative expression levels of cytochrome P450 (CYP450), glutathione S-transferase (GST) and apoptosis factor (CASP-3) were disturbed at different degrees after the stress. 3) The relative gene expression levels of the osmoregulation indexes such as ion transporters (VATP, NHE7, NKA-α, NKA-β and CA) and channel proteins (TIP4, CCP, CLC and AQP) occurred at different degrees of disorder after the stress. 4) The apoptosis index sucha as the relative expression level of apoptosis factor (CASP-3) gene decreased significantly in the three groups (P<0.05). Thus, it is inferred that nitrite and microplastic stress can induce the changes of immunity, detoxification metabolism and osmotic regulation in the gills of L. vannamei, affecting its normal physiological functions.
- Litopenaeus vannamei /
- Gill /
- Nitrite /
- Microplastics /
- Detoxification metabolism /
- Osmotic regulation

HTML全文

图 1 亚硝酸盐和微塑料胁迫对凡纳滨对虾鳃中氧化损伤指标的影响

注：数据上标不同字母表示组间差异显著 (P<0.05)；后图同此。

Figure 1. Effects of nitrite and microplastics stress on oxidative damage index in gills of L. vannamei

Note: Different letters on the bar indicate significant differences among the groups (P<0.05). The same case in the following figures.

下载: 全尺寸图片幻灯片

图 2 亚硝酸盐和微塑料胁迫对凡纳滨对虾鳃中抗氧化酶活性的影响

Figure 2. Effects of nitrite and microplastics stress on antioxidant enzymes activity in gills of L. vannamei

下载: 全尺寸图片幻灯片

图 3 亚硝酸盐和微塑料胁迫对凡纳滨对虾鳃组织免疫基因表达的影响

Figure 3. Effects of nitrite and microplastics stress on immune gene expression in gills of L. vannamei

下载: 全尺寸图片幻灯片

图 4 亚硝酸盐和微塑料胁迫对凡纳滨对虾鳃组织渗透调节基因表达的影响

Figure 4. Effects of nitrite and microplastics stress on osmotic regulation gene expression in gills of L. vannamei

下载: 全尺寸图片幻灯片

表 1 本研究所用引物序列

Table 1. Primer sequences used in this study

基因名称Gene name	正向引物 (5'—3')Forward primer (5'–3')	反向引物 (5'—3')Reverse primer (5'–3')
CASP-3	ATTATAGAGTGCCTGCGTTGCTACC	TGCGATATCATGCCTAGATGCTTGG
CYP450	CGCCGCCAAGGAAGTGAAGG	TGGGAGGCGATGGTGGTGTC
GST	ACGAACACTACGAACAGAAGGATGC	GCCAGGAAGTCGATGTAGGTTAGC
VATP	GCCAAGTATGAGCACCTAGCAGAC	TCGTCCACCACCTTACTGAAGAGAG
TIP4	GTCTCTCTGAACCCTTGCCAAACTC	GTTGCCTATGATTACCTGCGTCCTC
CCP	CCCTGAGACCTCTGTTTGCTGTG	TGGCAGTGTTCTTCGCATGTTCC
NHE7	CACCGCCATCCTCACCAAGTTC	ACAGAAGAGCACAGCCACAATACC
CLC	GCCGATAGGAGGTGTGTTGTTCAG	CGAAGAAGCCACGCCAGTAGTTC
NKA-α	TGAAATCGTGTTTGCCCGTACCTC	ACCATCACCAGTTACAGCCACAATG
NKA-β	GAACCCAGCCGACGAAGAATACG	CAGCAACAATAGGTGGCAGGTAGC
AQP	GCAGCCATCTTGAAGGGAGTGAC	ACGAGGACGAAGGTGATGAGGAG
CA	CCTATTCTGGCTCCCTCACTACCC	GTTCATCCTCTGGGCAACACTCG
β-actin	AGCTCATTGTAGAAGGTGTGATGCC	TCCTGACCCTGAAGTACCCCATTG

下载: 导出CSV

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