Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2023.02.002

The present experimental investigation was executed to evaluate the combined effect of probiotic bacterium Bacillus coagulans along with herb Mentha piperita on histological architecture; and extent of DNA damage after challenge trial with pathogenic bacterium Aeromonas hydrophila in Catla catla. In this experiment, six dietary treatments in triplicate were formulated; C0, M3 and M6 in which M. piperita was supplemented at 0, 3, 6 g kg⁻¹ of feed, while in dietary treatments CP, MP3 and MP6; supplementation of probiotic bacterium B. coagulans at the rate of 3000 CFU g⁻¹ was done along with different inclusion level of M. piperita (0, 3, 6 g kg⁻¹ of feed). Fishes were fed for 60 days on different dietary treatments. Growth performance, histological study and extent of DNA damage using comet assay was examined. Significantly (p < 0.05) high growth % gain in BW was observed in treatment MP6 followed by treatment M6. The histological analysis clearly revealed that dietary M. piperita without or with probiotic bacterium can protect the histoarchitecture of intestinal layers and liver even after challenge with A. hydrophila. Analysis of various comet parameters (tail length, olive moment, tail DNA, head DNA) affirmed that M. piperita supplemented diet can reduce the extent of DNA damage in fishes after challenge trial with A. hydrophila, however dietary supplementation of probiotic bacterium along with M. piperita can reduce the DNA damage up to larger extent confirming its combined protective effect of supplemented diets.

本实验研究旨在评估益生菌凝结芽孢杆菌和薄荷对猫鼬病原菌嗜水气单胞菌挑战试验后的组织学结构和 DNA 损伤程度的综合影响。在该实验中，共制定了六种一式三份的膳食处理：C0、M3 和 M6，其中蒌草的添加量分别为 0、3 和 6 g kg-1 饲料，而在 CP、MP3 和 MP6 膳食处理中，在添加不同水平的蒌草（0、3 和 6 g kg-1 饲料）的同时，还添加了 3000 CFU g-1 的益生菌凝结球菌。鱼类在不同的饮食处理下被喂养了 60 天。采用彗星试验对鱼类的生长性能、组织学研究和 DNA 损伤程度进行了检测。观察到 MP6 处理的体重增长百分比显著高于 M6 处理（p < 0.05）。组织学分析清楚地表明，即使在受到嗜水青虫的挑战后，不添加益生菌或添加益生菌的膳食中的蒌蒿能保护肠道层和肝脏的组织结构。对各种彗星参数（尾长、橄榄时刻、尾部 DNA、头部 DNA）的分析表明，补充瓜子黄酮的膳食可以减少鱼类在受到蚜蝇绥氏菌挑战后的 DNA 损伤程度，但在补充瓜子黄酮的同时补充益生菌可以在更大程度上减少 DNA 损伤，这证实了补充膳食的综合保护作用。

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引用次数: 0

CRISPR/Cas9-mediated mutation of Mstn confers growth performance in Culter alburnus juveniles CRISPR/Cas9 介导的 Mstn 基因突变可提高白腹角雉幼体的生长性能

Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2023.04.008

Myostatin is a member of the TGF-β superfamily and functions as a negative regulator for skeletal muscle development and growth. It has become the most targeted gene in aquaculture that used for selective breeding. Previous studies involved in genome editing in several fish species confirmed that CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats) system was highly efficient with lower off-target effect, however, no reports were raised in Culter alburnus. In this study, we employed CRISPR/Cas9 gene editing system to successfully disrupt mstn gene by co-injection with Cas9 protein and the targeted sgRNA in C. alburnus. Various Indel mutations were obtained with 82% knockout efficiency in the F0 generation by PCR sequencing. In addition, mutations in mstn that induced by CRISPR/Cas9 were detected in the F1 generation by individually mating the wild-type female with the F0 generation of mstn-KO male at sexual maturity. More importantly, the body weight and length were significantly elevated in mstn ^± group when compared to those of the control. As expected in mstn ^± group, the expression level of mstn was sharply reduced, whereas a slight increase was observed in two growth-related genes (myod and myog). Moreover, higher numbers of muscle fibers were observed in mstn ^± group, meaning that growth performance in mstn ^± individuals might be represented by increasing the number of muscle fibers. Taken together, our current study successfully obtained a site-specific modification of mstn using CRISPR/Cas9 technology, and these results provided a new insight for facilitating topmouth culter genetic studies and breeding.

Myostatin 是 TGF-β 超家族的成员，是骨骼肌发育和生长的负调控因子。它已成为水产养殖中用于选择性育种的最多目标基因。以往在多个鱼类物种中进行的基因组编辑研究证实，CRISPR/Cas9（Clustered Regularly Interspaced Short Palindromic Repeats）系统具有高效率、低脱靶效应的特点，但在白鲑鱼中尚未见报道。在本研究中，我们采用CRISPR/Cas9基因编辑系统，通过共注射Cas9蛋白和靶向sgRNA，成功地破坏了白僵菌的mstn基因。通过PCR测序，在F0代中获得了多种Indel突变，基因敲除效率高达82%。此外，通过将野生型雌性与F0代mstn-KO雄性在性成熟时单独交配，在F1代中检测到了CRISPR/Cas9诱导的mstn突变。更重要的是，与对照组相比，mstn±组的体重和体长明显增加。正如预期的那样，在mstn±组中，mstn的表达水平急剧下降，而两个与生长相关的基因（myod和myog）的表达水平则略有上升。此外，在 mstn ± 组中观察到了更多的肌纤维，这意味着 mstn ± 组个体的生长表现可能是通过增加肌纤维数量来体现的。综上所述，本研究利用CRISPR/Cas9技术成功地对mstn进行了位点特异性修饰，这些结果为促进顶口秆鱼的遗传研究和育种提供了新的见解。

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引用次数: 0

Simvastatin enhanced growth and altered testis transcriptome profiles of male tilapias 辛伐他汀促进雄性罗非鱼的生长并改变其睾丸转录组图谱

Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2023.05.010

Simvastatin (SIM), a commonly prescribed drug for lowering serum-lipid, has been detected in many aquatic environments. Exposure to high concentrations of SIM in rats and aquatic organisms resulted in toxicity to hepatocytes and negative effects on development. However, there is a paucity of studies on the effects of long-term exposure to low concentrations of SIM on aquaculture species. This study aimed to analyze the effects of environmentally relevant concentrations (0.1 μg/L and 1 μg/L) of SIM exposure for two months on developing Mozambique tilapias. Surprisingly, the growth of male tilapias was enhanced after a 2-month SIM exposure. The analysis of testis histology revealed that after the SIM exposure, the development of the testis was delayed, and sperm production was reduced. Sequencing of testis transcriptomes showed that the exposure to SIM altered the expression of many genes related to the development of testes and spermatogenesis. These data suggest that a 2-month exposure to environmentally relevant concentrations of increases the growth of male tilapias by inhibiting sexual development of males.

辛伐他汀（SIM）是一种用于降低血清脂质的常用处方药，在许多水生环境中都被检测到。大鼠和水生生物接触高浓度 SIM 会导致肝细胞中毒，并对发育产生负面影响。然而，有关长期接触低浓度 SIM 对水产养殖物种影响的研究却很少。本研究旨在分析环境相关浓度（0.1 μg/L 和 1 μg/L）的 SIM 接触两个月对莫桑比克罗非鱼发育的影响。令人惊讶的是，雄性罗非鱼在接触 SIM 2 个月后生长速度加快。睾丸组织学分析表明，接触 SIM 后，睾丸发育延迟，精子生成减少。睾丸转录组测序表明，接触 SIM 改变了许多与睾丸发育和精子生成有关的基因的表达。这些数据表明，暴露于环境相关浓度的 SIM 2 个月会抑制雄性罗非鱼的性发育，从而影响雄性罗非鱼的生长。

引用次数: 0

Ameliorative effect of walnut oil against cognitive impairment in alzheimers type dementia in rodent 核桃油对啮齿动物阿尔茨海默型痴呆认知功能障碍的改善作用

Q3 Agricultural and Biological Sciences

Oil Crop Science

Pub Date : 2024-11-01 DOI: 10.1016/j.ocsci.2024.09.003

Congcong Ma , Li Zhang , Lei Wang , Qingde Huang , Qianchun Deng , Fenghong Huang , Jiqu Xu

Walnut oil (WO), known for abundant polyunsaturated fatty acids and an array of bioactive substances such as tocopherols, phytosterols, squalene, melatonin, and polyphenols, which is endowed with numerous health advantages. The primary objective of this research was to ascertain the impact of WO on cognitive deficits in learning and memory impairment mice caused by scopolamine (SCOP). The Morris water maze and the step-down avoidance test were utilized to assess the memory and learning capabilities. WO notably counteracted the detrimental effects of SCOP on learning and memory in the Morris water maze, as indicated by a reduction in escape latency and swimming distance. Likewise, WO administration led to a notably reduced number of errors in training trial and an increased latency in testing trial when compared to the SCOP group in the step-down avoidance test,. Moreover, WO activated the cholinergic system of the brain by upregulating choline acetyltransferase activity and reducing acetylcholinesterase activity. These results suggest that WO has the potential to protect against memory decline in mice, offering a promising strategy for the prevention of memory-related disorders.

核桃油（WO）以丰富的多不饱和脂肪酸和一系列生物活性物质（如生育酚、植物甾醇、角鲨烯、褪黑激素和多酚）而闻名，具有众多健康优势。本研究的主要目的是确定 WO 对东莨菪碱（SCOP）导致的学习和记忆障碍小鼠认知缺陷的影响。研究采用了莫里斯水迷宫和降阶回避测试来评估小鼠的记忆和学习能力。在莫里斯水迷宫中，WO明显抵消了SCOP对学习和记忆的不利影响，这表现在逃逸潜伏期和游泳距离的缩短上。同样，在降阶回避测试中，与 SCOP 组相比，施用 WO 可显著减少训练试验中的错误次数，增加测试试验中的潜伏期。此外，WO 还能通过上调胆碱乙酰转移酶活性和降低乙酰胆碱酯酶活性来激活大脑的胆碱能系统。这些结果表明，WO 具有防止小鼠记忆力衰退的潜力，为预防记忆相关疾病提供了一种很有前景的策略。

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引用次数: 0

Design and practice of educational experiments on genetic epistasis. 遗传表观性教育实验的设计与实践。

Q3 Medicine

遗传

Pub Date : 2024-11-01 DOI: 10.16288/j.yczz.24-248

Yi Shi, Yao Yu, Yi-Lin Lü, Hong Lü

Genetic epistasis is a fundamental concept in genetics that describes how interactions between genes determine phenotypic traits. To enhance students' understanding and practical application of genetic epistasis, this experiment is designed and conducted using gene mutations in the adenine biosynthesis pathway of Saccharomyces cerevisiae (baker's yeast). S. cerevisiae is a classic model organism for genetic teaching experiments. In its adenine biosynthesis pathway, a mutation in the ADE2 gene leads to the accumulation of the intermediate 5'-phosphoribosylaminoimidazole (AIR), causing the cells to appear red. However, if a gene upstream of ADE2 in the adenine biosynthesis pathway (such as ADE8) is defective, the red phenotype of yeast will disappear. Conversely, a defect in a gene downstream of ADE2 (such as ADE1) does not alter the red phenotype. Therefore, ADE8 is epistatic to ADE2. In this experiment, the CRISPR-Cas9 genome editing technology is employed, allowing students to perform single knockout of ade2Δ, as well as double knockouts of ade2Δade8Δ and ade2Δade1Δ in S. cerevisiae. By observing the phenotypic changes in yeast mutants from white to red and back to white, students gain a profound understanding of the basic genetic theory of how genes determine phenotypes and the concept of epistasis in gene interactions. This experiment also enables students to master fundamental yeast genetic techniques, significantly enhancing their ability to design and conduct experiments in real research environments. This is of great significance for their future research work and academic development.

遗传表观性是遗传学中的一个基本概念，它描述了基因之间的相互作用如何决定表型性状。为了加强学生对遗传表观性的理解和实际应用，本实验设计并使用了酿酒酵母（面包酵母）腺嘌呤生物合成途径中的基因突变。酿酒酵母是遗传教学实验的经典模式生物。在其腺嘌呤生物合成途径中，ADE2 基因突变会导致中间体 5'- 磷酸核糖基氨基咪唑（AIR）的积累，从而使细胞呈现红色。但是，如果腺嘌呤生物合成途径中 ADE2 上游的一个基因（如 ADE8）出现缺陷，酵母的红色表型就会消失。相反，ADE2 下游基因（如 ADE1）的缺陷不会改变红色表型。因此，ADE8 是 ADE2 的外显子。在本实验中，学生利用 CRISPR-Cas9 基因组编辑技术，对 S. cerevisiae 进行了 ade2Δ 的单基因敲除，以及 ade2Δade8Δ 和 ade2Δade1Δ 的双基因敲除。通过观察酵母突变体从白色到红色再到白色的表型变化，学生深刻理解了基因决定表型的基本遗传理论和基因相互作用中的表观遗传概念。本实验还使学生掌握了基本的酵母遗传技术，大大提高了他们在实际研究环境中设计和进行实验的能力。这对他们今后的研究工作和学术发展具有重要意义。

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引用次数: 0

Identification and functional characterization of CD209 homologous genes in zebrafish. 斑马鱼 CD209 同源基因的鉴定和功能表征

Q3 Medicine

遗传

Pub Date : 2024-11-01 DOI: 10.16288/j.yczz.24-181

Xiao-Jun Yang, Zhen-Han Huang, Wei Liu, Wen-Qing Zhang, Zhi-Bin Huang

Innate immune responses play a crucial role in maintaining homeostasis, their initiation closely related to pattern recognition receptors or damage-associated molecules on the surface of innate immune cells. CD209, a pattern recognition receptor on the surface of macrophages or dendritic cells, plays an important role in immune functions. However, the impact of CD209 on innate immune cells such as macrophages or neutrophils in vivo remains unclear. In this study, through multiple sequence alignment and phylogenetic tree construction, three genes homologous to human CD209 were found in zebrafish. These are cd209(Ensembl ID:ENSDARG00000029461), zgc:174904(Ensembl ID:ENSDARG00000059049) and si:dkey-187I7.2(Ensembl ID: ENSDARG00000096624).Compared to the cd209 and si:dkey-187i8.2 genes in the Ensembl database, zgc:174904 is more similar to human CD209 in sequence. Using whole-mount in situ hybridization and fluorescence co-localization experiments, it was found that zgc:174904 is mainly expressed in macrophages. Further morpholino knockdown experiments showed that knocking down zgc:174904 leads to an upregulation of M1-type macrophage-related genes and a decrease in the number of mature neutrophils, indicating that zgc:174904 is functionally more similar to CD209. These findings not only reveal the potential role of CD209 in regulating macrophage function and neutrophil development but also provide significant insights for research into the mechanisms of innate immunity.

先天性免疫反应在维持体内平衡方面发挥着至关重要的作用，其启动与先天性免疫细胞表面的模式识别受体或损伤相关分子密切相关。CD209 是巨噬细胞或树突状细胞表面的一种模式识别受体，在免疫功能中发挥着重要作用。然而，CD209 在体内对巨噬细胞或中性粒细胞等先天性免疫细胞的影响仍不清楚。本研究通过多序列比对和系统发生树的构建，在斑马鱼中发现了三个与人类 CD209 同源的基因。与Ensembl数据库中的cd209和si:dkey-187i8.2基因相比，zgc:174904与人类CD209的序列更为相似。通过整装原位杂交和荧光共定位实验发现，zgc:174904 主要在巨噬细胞中表达。进一步的吗啉敲除实验表明，敲除zgc:174904会导致M1型巨噬细胞相关基因的上调和成熟中性粒细胞数量的减少，这表明zgc:174904在功能上与CD209更为相似。这些发现不仅揭示了CD209在调控巨噬细胞功能和中性粒细胞发育中的潜在作用，而且为先天性免疫机制的研究提供了重要启示。

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引用次数: 0

Stability analysis of reef fish communities based on symbiotic graph model 基于共生图模型的珊瑚礁鱼类群落稳定性分析

Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2023.03.005

The community stability of coral reefs and fish is the focus of ecological monitoring of coral reefs. Among them, the realization of effective metrics of variations in reef fish communities (i.e., the combined communities of coral reefs and fish) is important for analyzing the stability of communities as well as maintaining the ecological balance of coral reefs. Based on coral reef and fish data collected at St. John's Island from 2004 to 2010, this study proposes a symbiotic graph modeling method to express the biological relationships of reef fish communities, and a Pyramid Match graph kernel method for fusing Attributes (PMA) to quantify community fluctuations to measure interannual variability of communities. The results showed that the community similarity was low in 2006, 2007, and 2008. The total coral cover rate in the study area decreased by 32.04% from 2006 to 2007 and increased by 24% in 2008. The total number of fish fell from 3780 in 2006 to 2596 in 2007 and rose to 6249 in 2008. Among them, the proportion of herbivorous fish decreased to 30.84% in 2007. Furthermore, we have combined the Louvain algorithm with the proposed PMA method to effectively identify the regions that should be prioritized for protection. Experiments were conducted on real datasets with good results, demonstrating the potential of the proposed method to assist in the analysis of community stability and identification of priority conservation areas.

珊瑚礁和鱼类的群落稳定性是珊瑚礁生态监测的重点。其中，实现珊瑚礁鱼类群落（即珊瑚礁和鱼类的组合群落）变化的有效度量，对于分析群落的稳定性以及维持珊瑚礁的生态平衡具有重要意义。本研究基于 2004 年至 2010 年在圣约翰岛采集的珊瑚礁和鱼类数据，提出了表达珊瑚礁鱼类群落生物关系的共生图建模方法和量化群落波动的融合属性金字塔匹配图核方法（PMA），以衡量群落的年际变异性。结果表明，2006 年、2007 年和 2008 年的群落相似度较低。研究区域的珊瑚总覆盖率从 2006 年到 2007 年下降了 32.04%，2008 年上升了 24%。鱼类总数从 2006 年的 3780 条下降到 2007 年的 2596 条，2008 年又上升到 6249 条。其中，草食性鱼类的比例在 2007 年下降到 30.84%。此外，我们还将卢万算法与所提出的 PMA 方法相结合，有效地确定了应优先保护的区域。在真实数据集上进行的实验取得了良好的效果，证明了所提出的方法在协助分析群落稳定性和确定优先保护区域方面的潜力。

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引用次数: 0

Effects of biological flocculation technology (BFT) on water quality dynamics and immune response of grass carp (Ctenopharyngodon idella) 生物絮凝技术（BFT）对草鱼水质动态和免疫反应的影响

Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2023.04.005

The biofloc technology (BFT) offers benefits in improving feed utilization and reducing the feed conversion ratio compared to the recirculating aquaculture system (RAS). In this study, high-throughput 16S rRNA gene sequencing was employed to investigate and compare the bacterial communities in these aquaculture systems and the gut microbiota of grass carp reared in them. We observed a significant distinction between the microbial communities of the aquaculture systems and the intestinal microbiota of grass carp. The genera cetobacterium and bacillus were more abundant in the BFT system, accounting for only 0.05% and 0.25% in the RAS. Our study also demonstrated that BFT could influence the intestinal microorganisms of grass carp (Ctenopharyngodon Idella) by reducing the relative abundance of burkholderia-caballeronia-paraburkholderia and increasing that of cetobacterium. Based on bugBase community predictions, the potential pathogenicity in BFT and grass carp intestine was lower than in RAS. Regarding gene expression in the grass carp intestine, immunosuppressive genes showed higher expression, while pro-inflammatory genes exhibited lower expression in grass carp reared in BFT. Furthermore, the final average length and weight of grass carp in the BFT system were significantly higher than those in the RAS. This study provides valuable insights into gut microbiome ecology in relation to two grass carp aquaculture systems, which could be effectively applied in grass carp management to promote health and improve water quality.

与循环水养殖系统（RAS）相比，生物絮团技术（BFT）在提高饲料利用率和降低饲料转化率方面具有优势。本研究采用高通量 16S rRNA 基因测序技术，研究并比较了这些水产养殖系统中的细菌群落以及在其中饲养的草鱼的肠道微生物群落。我们观察到水产养殖系统的微生物群落与草鱼肠道微生物群落之间存在明显差异。在 BFT 系统中，鲸杆菌属和芽孢杆菌属的数量更多，而在 RAS 系统中仅占 0.05% 和 0.25%。我们的研究还表明，BFT 可以通过降低burkholderia-caballeronia-paraburkholderia 的相对丰度和提高 cetobacterium 的相对丰度来影响草鱼（Ctenopharyngodon Idella）的肠道微生物。根据 bugBase 群落预测，BFT 和草鱼肠道的潜在致病性低于 RAS。在草鱼肠道基因表达方面，BFT饲养草鱼的免疫抑制基因表达较高，而促炎基因表达较低。此外，BFT 系统中草鱼的最终平均体长和体重明显高于 RAS 系统。本研究提供了有关两种草鱼养殖系统的肠道微生物组生态学的宝贵见解，可有效地应用于草鱼管理，以促进健康和改善水质。

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引用次数: 0

Effects of microbe-derived antioxidants on growth, digestive and aminotransferase activities, and antioxidant capacities in the hepatopancreas of Eriocheir sinensis under ammonia nitrogen stress 微生物源抗氧化剂对氨氮胁迫下中华鳖肝胰腺的生长、消化酶和转氨酶活性以及抗氧化能力的影响

Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2022.12.002

This study evaluated the effects of feeding a microbial-derived antioxidant diet (MA) for 56 days on the growth performance of Eriocheir sinensis, as well as on digestive and metabolic enzyme activity and antioxidant capacity under ammonia stress (400 mg/L ammonia nitrogen for 48 h). The weight gain at 28d and the specific growth rate at 56ds for crabs in the MA group were significantly higher than those in the control diet (C) group (p < 0.05). Compared with the C group, the color of the hepatopancreas in the MA group was significantly improved (p < 0.05). A significant decrease in the level of Hemolymph ammonia was found in the microbial antioxidant ammonia-N (MA_A) group (p < 0.05), which may be related to the decrease of transaminase (AST, ALT, GDH) activities caused by MA diet (p < 0.05). There is a significant increase in the digestive enzyme activities (Trypsin and Lipase), which reduced the accumulation of triglycerides in the hepatopancreas in the (MA_A) group (p < 0.05). In addition, the antioxidant damage of crabs in hepatopancreas caused by ammonia nitrogen toxicity could also significantly be improved by MA fed in the (MA_A) group (p < 0.05). microbe-derived antioxidants (supplemented with 0.2% in feeds) can protect the health of hepatopancreas, promote growth, and improve the ability to resist ammonia nitrogen stress for E.sinensis.

本研究评估了投喂微生物源抗氧化饲料（MA）56天对中华绒螯蟹生长性能的影响，以及在氨胁迫（400 mg/L氨氮，48 h）条件下对消化代谢酶活性和抗氧化能力的影响。MA组螃蟹28 d的增重和56 d的特定生长率显著高于对照组（P <0.05）。与 C 组相比，MA 组的肝胰脏颜色明显改善（p < 0.05）。微生物抗氧化氨-N（MA_A）组的血淋巴氨水平明显下降（p <0.05），这可能与 MA 日粮引起的转氨酶（AST、ALT、GDH）活性下降有关（p <0.05）。消化酶活性（胰蛋白酶和脂肪酶）明显增加，减少了（MA_A）组肝胰腺中甘油三酯的积累（p < 0.05）。此外，(MA_A)组螃蟹肝胰腺因氨氮毒性引起的抗氧化损伤也可通过投喂 MA 得到显著改善（p < 0.05）。微生物衍生的抗氧化剂（在饲料中添加 0.2%）可保护中华绒螯蟹肝胰腺健康，促进生长，提高其抵抗氨氮胁迫的能力。

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引用次数: 0

IKKε positively regulates NF-κB, MAPK, and IRF3-mediated type I IFN signaling pathways in Japanese eel (Anguilla japonica) IKKε 正向调节日本鳗鲡体内 NF-κB、MAPK 和 IRF3 介导的 IFN 信号通路

Q1 Agricultural and Biological Sciences

Aquaculture and Fisheries

Pub Date : 2024-11-01 DOI: 10.1016/j.aaf.2023.01.001

IKKε is an IκB kinase participating in the control of NF-κB and type I IFN signal pathways in mammals. However, the function of IKKε in regulating immune response is largely unknown in teleost. Herein, an IKKε homologue named AjIKKε was characterized in Japanese eel (Anguilla japonica). AjIKKε has an N-terminal kinase domain, a ubiquitin-like domain, and a coiled coil-containing domain (CC), which is conserved and similar to its counterpart in mammals. Expression analysis showed that AjIKKε could be up-regulated in kidney, spleen, and particularly in liver under the stimulation of poly I:C, LPS, and Aeromonas hydrophila infection. In vitro, the mRNA levels of AjIKKε were significantly provoked in eel liver cells stimulated by LPS and poly I:C, or the different concentrations of A. hydrophila. The overexpression of AjIKKε could not only induce a significantly higher level of promoter activity of human NF-κB, AP-1, and IFN-β in a dose-dependent manner but also up-regulate the activation of promoters of Japanese eel cRel, AP1, IL6, IFN4, IRF3, and IRF7 in HEK293 cells. RNAi studies showed that after AjIKKε was knocked down, the expression levels of IL1, IL6, TNFα, c-Jun, IFN2, IFN3, MX1, MX2, and IRF3 genes were significantly down-regulated in liver, spleen, and kidney of Japanese eels. In addition, the mutants of AjIKKε-K39A, AjIKKε-S174A, and AjIKKε-ΔCC failed to activate Japanese eel IFN4, IRF3 and human IFN-β promoters in HEK293 cells. Collectively, these results suggest that AjIKKε may function as a positive regulator of NF-κB, MAPK, and IRF3-mediated type I IFN signaling pathways related to immune response evoked by bacterial and viral infection.

IKKε是一种IκB激酶，参与控制哺乳动物的NF-κB和I型IFN信号通路。然而，IKKε在远洋鱼类中调节免疫反应的功能尚不清楚。在此，研究人员鉴定了日本鳗鲡（Anguilla japonica）中名为AjIKKε的IKKε同源物。AjIKKε具有一个N-末端激酶结构域、一个泛素样结构域和一个含线圈的结构域（CC），这些结构域与哺乳动物中的同源物保守且相似。表达分析表明，在多聚 I:C、LPS 和嗜水气单胞菌感染的刺激下，AjIKKε可在肾脏、脾脏，特别是肝脏中上调。在体外，鳗鱼肝细胞在 LPS 和 poly I:C 或不同浓度的嗜水气单胞菌的刺激下，AjIKKε的 mRNA 水平显著升高。在 HEK293 细胞中，过表达 AjIKKε 不仅能以剂量依赖的方式诱导人 NF-κB、AP-1 和 IFN-β 启动子活性水平的显著提高，还能上调日本鳗鲡 cRel、AP1、IL6、IFN4、IRF3 和 IRF7 启动子的活化。RNAi研究表明，敲除AjIKKε后，日本鳗鱼肝脏、脾脏和肾脏中IL1、IL6、TNFα、c-Jun、IFN2、IFN3、MX1、MX2和IRF3基因的表达水平显著下调。此外，AjIKKε-K39A、AjIKKε-S174A和AjIKKε-ΔCC突变体在HEK293细胞中不能激活日本鳗鱼IFN4、IRF3和人IFN-β启动子。这些结果表明，AjIKKε可能是NF-κB、MAPK和IRF3介导的I型IFN信号通路的正调控因子，与细菌和病毒感染诱发的免疫反应有关。

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引用次数: 0

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