低氧胁迫对军曹鱼幼鱼免疫相关基<italic/>因转录水平表达的影响

王维政; 杨林桐; 杨二军; 谢瑞涛; 陈刚; 黄建盛

doi:10.12284/hyxb2021116

低氧胁迫对军曹鱼幼鱼免疫相关基因转录水平表达的影响

doi: 10.12284/hyxb2021116

王维政^1,,
杨林桐¹,
杨二军¹,
谢瑞涛^{1, 3},
陈刚^{1, 2},
黄建盛^{1, 2, ,}

1.
广东海洋大学水产学院，广东湛江 524088
2.
南方海洋科学与工程广东省实验室（湛江），广东湛江 524025
3.
广东恒兴饲料实业股份有限公司，广东湛江 524022

基金项目: 财政部和农业农村部：国家现代农业产业技术体系资助项目（CARS-47）；南方海洋科学与工程广东省实验室(湛江)资助项目(ZJW-2019-06)

详细信息

作者简介:
王维政(1995-)，男，广东省广州市人，主要研究方向为海水鱼类养殖生理生态学。E-mail：1029588896@qq.com

通讯作者:
黄建盛，副教授，主要研究方向海水鱼养殖生理生态学。E-mail：huangjs@gdou.edu.cn

中图分类号: Q786
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出版历程
- 收稿日期: 2021-01-30
- 修回日期: 2021-04-12
- 网络出版日期: 2021-05-12
- 刊出日期: 2021-09-25

Transcription level of immune related genes of juvenile cobia (Rachycentron canadum) under hypoxia stress

Wang Weizheng^1
,,
Yang Lintong¹,
Yang Erjun¹,
Xie Ruitao^{1, 3},
Chen Gang^{1, 2},
Huang Jiansheng^{1, 2
, ,}

1.
Fishery College, Guangdong Ocean University, Zhanjiang 524088, China
2.
Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang 524025, China
3.
Guangdong Evergreen Feed Industry Co. Ltd., Zhanjiang 524022, China

摘要

摘要: 为研究低氧胁迫对军曹鱼幼鱼免疫功能的影响，将幼鱼暴露于溶氧浓度为(3.15±0.21) mg/L的水体28 d，测定不同时间点肿瘤坏死因子α (TNFα)、肿瘤坏死因子α诱导蛋白3 (TNFAIP3)、白细胞介素1β (IL-1β)、白细胞介素1受体2 (IL-1R2)、白细胞介素17C (IL-17C)和热休克蛋白70 (HSP70)等免疫相关基因在幼鱼鳃、肝脏、肠道和脾脏中的转录水平表达量。结果显示：(1)在幼鱼的鳃中，TNFα 和 IL-1R2 基因转录水平表达量在胁迫1 d和14 d时极显著低于对照组(p<0.01)，胁迫 28 d则分别表现为极显著(p<0.01)和显著下降(p<0.05)；TNFAIP3在胁迫1 d时极显著升高(p<0.01)，在胁迫7 d和14 d时则分别表现为显著(p<0.05)和极显著(p<0.01)下降；IL-1β在胁迫1 d时极显著下降(p<0.01)后极显著升高 (p<0.01)；IL-17C在胁迫7 d和14 d时极显著下降(p<0.01)；HSP70 在胁迫1 d时显著下降(p<0.05)，在胁迫14 d和28 d时均极显著降低(p<0.01)；(2)在肝脏中，TNFα 和 IL-1R2 基因转录水平表达量在胁迫1 d和28 d时显著下降(p<0.05)，在胁迫14 d时则极显著降低(p<0.01)；TNFAIP3在胁迫1 d时极显著上升(p<0.01)，在胁迫14 d和28 d时则极显著下降(p<0.01)；IL-1β在所有胁迫时间点均极显著上升(p<0.01)；IL-17C 在胁迫7 d、14 d和28 d时均极显著低于对照水平(p<0.01)；HSP70基因表达量持续上升，并于胁迫28 d时达到顶峰(p<0.01)；(3)在肠道中，TNFα、IL-1β、IL-1R2、IL-17C和HSP70基因转录水平表达量在胁迫的所有时间点均极显著高于对照组(p<0.01)，TNFAIP3在胁迫1 d时表达量极显著升高(p<0.01)后下降，在胁迫7 d和14 d时分别与对照组具有显著(p<0.05)和极显著差异(p<0.01)；(4)在脾脏中，TNFα和IL-17C 基因转录水平表达量在胁迫1 d、7 d和14 d时极显著下降(p<0.01)，在胁迫28 d时则显著降低(p<0.05)；IL-1β在胁迫1 d和28 d时极显著下降(p<0.01)，在胁迫7 d和14 d时则显著降低(p<0.05)；IL-1R2在胁迫1 d、7 d和28 d时极显著下降(p <0.01)，在胁迫14 d时则显著下降(p<0.05)；HSP70基因表达量在胁迫1 d、7 d和28 d时均极显著下降(p<0.01)。研究结果表明，28 d的低氧胁迫后，军曹鱼幼鱼免疫基因的转录表达水平发生显著性变化，预示长时间的低氧可能抑制了军曹鱼的免疫功能，引发肠道炎症，并增加了军曹鱼感染病原菌的风险。
- 军曹鱼 /
- 低氧 /
- 免疫基因
Abstract: To study the effect of hypoxia on immune function of juvenile cobia (Rachycentron canadum), the transcription level of immune related genes were detected in gills, liver, intestines, and spleen after exposing to (3.15±0.21) mg/L hypoxia stress for 28 d. The results showed: (1) in gills, the transcription level of tumor necrosis factor α (TNFα) and type 2 interleukin-1 receptor (IL-1R2) extremely decreased (p<0.01) on 1 d and 14 d of stress. On 28 d of stress, TNFα extremely decreased (p<0.01) while IL-1R2 significantly decreased (p<0.05). Tumor necrosis factor alpha induced protein 3 (TNFAIP3) extremely increased (p<0.01) on 1 d and then significantly decreased (p<0.05) on 7 d and extremely decreased (p<0.01) on 14 d. Interleukin-1β (IL-1β) extremely decreased (p<0.01) on 1 d and then extremely increased (p<0.01) . Interleukin-17C (IL-17C) extremely decreased (p<0.01) on 7 d and 14 d, while heat shock protein 70 (HSP70) significantly decreased (p<0.05) on 1 d and extremely decreased (p<0.01) on 14 d and 28 d. (2) In liver, the transcription level of TNFα and IL-1R2 significantly decreased (p<0.05) on 1 d and 28 d and extremely decreased (p<0.01) on 14 d. TNFAIP3 extremely increased (p<0.01) on 1 d and extremely decreased (p<0.01) on 14 d and 28 d. IL-1β extremely increased (p<0.01) at all stress time points, while IL-17C extremely decreased (p<0.01) on 7 d, 14 d and 28 d. HSP70 continued to increase and reached the maximum on 28 d (p<0.01). (3) In the intestine, the transcription level of TNFα, IL-1β, IL-1R2, IL-17C and HSP70 genes were higher (p<0.01) than those in the control group at all stress time points. TNFAIP3 extremely increased (p<0.01) on 1 d and significantly decreased (p<0.05) on 7 d and extremely decreased (p<0.01) on 14 d. (4) In spleen, the transcription level of TNFα and IL-17C extremely decreased (p<0.01) on 1 d, 7 d, 14 d and significantly decreased (p<0.05) on 28 d. IL-1β extremely decreased (p<0.01) on 1 d, 28 d and significantly decreased (p<0.05) on 7 d and 14 d. IL-1R2 extremely decreased (p<0.01) on 1 d, 7 d, 28 d and significantly decreased (p<0.05) on 14 d, while HSP70 extremely decreased (p<0.01) on 1 d, 7 d and 28 d. The results indicated that immune genes of transcription level in juvenile cobia showed significantly change after 28 d of hypoxia stress, suggesting that long-term hypoxia may inhibit the immune function, cause intestinal inflammation, and increase the risk of pathogen infection in cobia.
- Rachycentron canadum /
- hypoxia /
- immune gene

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图 1 低氧胁迫后军曹鱼幼鱼鳃组织TNFα（a）、TNFAIP3（b）、IL-1β（c）、IL-1R2（d）、IL-17C（e）和HSP70（f）基因的相对表达量

*表示显著差异（p<0.05），**表示极显著差异（p<0.01）

Fig. 1 Relative expression of TNFα (a), TNFAIP3 (b), IL-1β (c), IL-1R2 (d), IL-17C (e) and HSP70 (f) genes in gill of juvenile cobia after hypoxia

*Shows significant difference (p<0.05), **shows extremely significant difference (p<0.01)

下载: 全尺寸图片幻灯片

图 2 低氧胁迫后军曹鱼幼鱼肝组织TNFα（a）、TNFAIP3（b）、IL-1β（c）、IL-1R2（d）、IL-17C（e）和HSP70（f）基因的相对表达量

*表示显著差异（p<0.05），**表示极显著差异（p<0.01）

Fig. 2 Relative expression of TNFα (a), TNFAIP3 (b), IL-1β (c), IL-1R2 (d), IL-17C (e) and HSP70 (f) genes in liver of juvenile cobia after hypoxia

*Shows significant difference (p<0.05), **shows extremely significant difference (p<0.01)

下载: 全尺寸图片幻灯片

图 3 低氧胁迫后军曹鱼幼鱼肠组织TNFα（a）、TNFAIP3（b）、IL-1β（c）、IL-1R2（d）、IL-17C（e）和HSP70（f）基因的相对表达量

*表示显著差异（p<0.05），**表示极显著差异（p<0.01）

Fig. 3 Relative expression of TNFα (a), TNFAIP3 (b), IL-1β (c), IL-1R2 (d), IL-17C (e) and HSP70 (f) genes in intestine of juvenile cobia after hypoxia

*Shows significant difference (p<0.05), **shows extremely significant difference (p<0.01)

下载: 全尺寸图片幻灯片

图 4 低氧胁迫后军曹鱼幼鱼脾组织TNFα（a）、TNFAIP3（b）、IL-1β（c）、IL-1R2（d）、IL-17C（e）和HSP70（f）基因的相对表达量

*表示显著差异（p<0.05），**表示极显著差异（p<0.01）

Fig. 4 Relative expression of TNFα (a), TNFAIP3 (b), IL-1β (c), IL-1R2 (d), IL-17C (e) and HSP70 (f) genes in spleen of juvenile cobia after hypoxia

*Shows significant difference (p<0.05), **shows extremely significant difference (p<0.01)

下载: 全尺寸图片幻灯片

表 1 本研究所用引物序列

Tab. 1 Primer sequences used in this study

引物名称	序列(5’−3’)	用途
TNFα-F	GCTGGAGTGGAAGAACGGTCAA	qRT-PCR
TNFα-R	CTTGCCGCCTATGGAGTCTGAG	qRT-PCR
TNFAIP3-F	TCCAGAATGCCTCACGGTGTC	qRT-PCR
TNFAIP3-R	AGTGTGGTGTGCGATGTGTCA	qRT-PCR
IL-1β-F	GCAGAGAATCGGCACAGACA	qRT-PCR
IL-1β-R	GAAGGTTCGGTAGCGGTTGT	qRT-PCR
IL-1R2-F	ACCAAGGAGGAAGACACGAAGT	qRT-PCR
IL-1R2-R	TGCCGAGCCAGGATGTAATCA	qRT-PCR
IL-17C-F	GCCAAAGAAATGCGAGGAGATG	qRT-PCR
IL-17C-R	ACAGAGGCACTCAGCGAAGA	qRT-PCR
HSP70-F	TCACTGGAGTCCTACGCCTAC	qRT-PCR
HSP70-R	GCTGATGCTCATACTCGTCCTT	qRT-PCR
β-actin-F	AGGGAAATTGTGCGTGAC	qRT-PCR
β-asctin-R	AGGCAGCTCGTAGCTCTT	qRT-PCR

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