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Potential anticancer activity analysis of piscidin 5-like from larimichthys crocea

Acta Oceanologica Sinica 2022年第3期41卷 53-60页

作者： Libing Zheng Jiayin Qiu Huihui Liu Changfeng Chi Longshan Lin National and Provincial Joint Laboratory of Exploration and Utilization of Marine Aquatic Genetic Resources School of Marine Science and TechnologyZhejiang Ocean UniversityZhoushan 316022China Third Institute of Oceanography Ministry of Natural ResourcesXiamen 361005China

Antitumor activity is one characteristic function of some certain antimicrobial peptides(AMPs)found in recent years.In the present study,we attempted to detect potential anticancer activity of a recombinant piscidin 5-like from larimichthys crocea(r Lc-P5L)which owned widely antibacterial and strong antiparasitic activity in vitro.The light microscope observation indicated r Lc-P5L was of antitumor activity to He La cells,293 T cells and L929 cells.MTT assay showed the toxic sensitivity of r Lc-P5L to three tumor cell strains was 293 T>L929>He La.Scanning electron microscope(SEM)results showed r Lc-P5L behaved like a lytic peptide to cause damage on cells membrane of L929 cells by forming globular clusters,even pores at 60μmol/L,or degrading membrane to make it completely lose cytoskeleton structure at 80μmol/L;r Lc-P5L treatment also resulted in DNA degradation.Fluorescence observation results indicated r Lc-P5L could cause L929 cells at least two obvious changes:one is nucleus,nuclear chromatin condensed in the margin,nuclear volume became smaller and shrank to be out of shape,or lysed to be debris;the other is cytoskeleton,they became disordered and polarized to make cells atrophic shapes,or even lysed to be debris.In summary,r Lc-P5L owned potential anticancer activity causing membrane structure damage and genome DNA degradation.Interestingly,treatment with different concentration of r Lc-P5L seemingly caused the similar but different changes,whether it indeed gave rise to cancer cells diverse death way,the further studies should be performed,and the detailed mechanisms were still need further explored.

关键词： larimichthys crocea piscidin 5-like anticancer activity membrane destruction DNA degradation

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Morphological changes and variations in Na+/K+-ATPase activity in the gills of juvenile large yellow croaker (larimichthys crocea) at low salinity

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Aquaculture and Fisheries 2022年第3期7卷 313-320页

作者：滕广亮黄伟卿吉成龙陈云龙单秀娟 College of Fisheries and Life Science Shanghai Ocean UniversityShanghai201306China Laboratory for Marine Fisheries Science and Food Production Processes Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China Yellow Sea Fisheries Research Institute Chinese Academy of Fishery ScienceQingdao266071China College of Life Science Ningde Normal UniversityNingde352100China Ningde Dingcheng Fishery Company Limited Ningde352000China Yantai Institute of Coastal Zone Research Chinese Academy of SciencesYantai264003China

Gill morphological changes and physiological responses in juvenile large yellow croaker(larimichthys crocea)were examined upon exposure to low salinity after indoor culture of the fish at salinities of 2,4,6,8,and 24‰(control group).The thickness of the lamellae was significantly higher in the low-salinity groups than in the control group;in contrast,the interlamellar space was significantly lower in the low-salinity groups than in the control group.Additionally,a significant negative correlation was found between the thickness of lamellae and interlamellar spaces(P<0.01).Mitochondria-rich cells(MRCs)were mainly found in the filament at 24‰salinity and proliferated in the lamellae at lower salinities,suggesting that filament and lamellar MRCs are responsible for ion secretion and absorption,respectively.Meanwhile,the activity of Na+/K+-ATPase(NKA)was significantly elevated with the decrease in salinity from 24‰to 4‰(P<0.05),which was consistent with MRCs proliferation.Finally,the activity of NKA declined at 2‰salinity(P<0.05),indicating the limit of osmoregulation,which was consistent with the degeneration and necrosis of the lamellae.Additionally,different levels of gill histopathological lesions,including pavement cell(PVC)exfoliation,lamellar epithelial lifting,edema,fusion,aneurism,and necrosis,were observed from salinities of 24 to 4‰,likely reducing the respiratory efficiency and compromising the health of juvenile fish.In conclusion,large yellow croaker juveniles could improve the osmoregulatory capacity by increasing lamellar MRCs and NKA activity with the decrease in salinity from 24 to 4‰.However,the associated histopathological lesions are likely to negatively influence the fish by affecting respiration and osmoregulation,especially when the salinity is below 4‰.

关键词： larimichthys crocea Osmoregulation Gill morphology Histopathological lesions Low salinity

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Hif-1α/Hsf1/Hsp70 signaling pathway regulates redox homeostasis and apoptosis in large yellow croaker(larimichthys crocea)under environmental hypoxia

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Zoological Research 2021年第6期42卷 746-760页

作者： Sheng-Yu Luo Jing-Qian Wang Cheng Liu Xin-Ming Gao Yi-Bo Zhang Jie Ding Cong-Cong Hou Jun-Quan Zhu Bao Lou Wei-Liang Shen Xiong-Fei Wu Chun-Dan Zhang Dao-Jun Tang Key Laboratory of Applied Marine Biotechnology by the Ministry of Education School of Marine SciencesNingbo UniversityNingboZhejiang 315211China Zhejiang Academy of Agricultural Sciences HangzhouZhejiang 310021China State Key Laboratory of Large Yellow Croaker Breeding Ningbo Academy of Oceanology and FisheryNingboZhejiang 315012China

Oxygen is an essential molecule for animal respiration,growth,and survival.Unlike in terrestrial environments,contamination and climate change have led to the frequent occurrence of hypoxia in aquatic environments,thus impacting aquatic animal survival.However,the adaptative mechanisms underlying fish responses to environmental hypoxia remain largely unknown.Here,we used large yellow croaker(larimichthys crocea)and large yellow croaker fry(LYCF)cells to investigate the roles of the Hif-1α/Hsf1/Hsp70 signaling pathway in the regulation of cellular redox homeostasis,and apoptosis.We confirmed that hypoxia induced the expression of Hif-1α,Hsf1,and Hsp70 in vivo and in vitro.Genetic Hsp70 knockdown/overexpression indicated that Hsp70 was required for maintaining redox homeostasis and resisting oxidative stress in LYCF cells under hypoxic stress.Hsp70 inhibited caspase-dependent intrinsic apoptosis by maintaining normal mitochondrial membrane potential,enhancing Bcl-2 mRNA and protein expression,inhibiting Bax and caspase3 mRNA expression,and suppressing caspase-3 and caspase-9 activation.Hsp70 suppressed caspaseindependent intrinsic apoptosis by inhibiting nuclear translocation of apoptosis-inducing factor(AIF)and disturbed extrinsic apoptosis by inactivating caspase-8.Genetic knockdown/overexpression of Hif-1αand dual-luciferase reporter assay indicated that Hif-1αactivated the Hsf1 DNA promoter and enhanced Hsf1 mRNA transcription.Hsf1 enhanced Hsp70 mRNA transcription in a similar manner.In summary,the Hif-1α/Hsf1/Hsp70 signaling pathway plays an important role in regulating redox homeostasis and anti-apoptosis in L.crocea under hypoxic stress.

关键词： Hypoxia larimichthys crocea Apoptosis Redox homeostasis Hif-1α/Hsf1/Hsp70

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Novel insights into host-pathogen interactions of large yellow croakers(larimichthys crocea)and pathogenic bacterium Pseudomonas plecoglossicida using time-resolved dual RNA-seq of infected spleens

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Zoological Research 2020年第3期41卷 314-327页

作者： Yi Tang Ge Xin Ling-Min Zhao Li-Xing Huang Ying-Xue Qin Yong-Quan Su Wei-Qiang Zheng Bin Wu Nan Lin Qing-Pi Yan Fisheries College Key Laboratory of Healthy Mariculture for the East China SeaMinistry of AgricultureJimei UniversityXiamenFujian 361021China State Key Laboratory of Large Yellow Croaker Breeding Ningde Fufa Aquatic Products Co.Ltd.NingdeFujian 352000China Fujian Provincial Fishery Technical Extention Center FuzhouFujian 350003China

Host-pathogen interactions are highly complex,involving large dynamic changes in gene expression during infection. These interactions are fundamental to understanding anti-infection immunity of hosts, as well as the pathogenesis of pathogens. For bacterial pathogens interacting with animal hosts, timeresolved dual RNA-seq of infected tissue is difficult to perform due to low pathogen load in infected tissue. In this study, an acute infection model of larimichthys crocea infected by Pseudomonas plecoglossicida was established. The spleens of infected fish exhibited typical symptoms, with a maximum bacterial load at two days post-injection(dpi). Time-resolved dual RNA-seq of infected spleens was successfully applied to study hostpathogen interactions between L. crocea and P.plecoglossicida. The spleens of infected L. crocea were subjected to dual RNA-seq, and transcriptome data were compared with those of noninfected spleens or in vitro cultured bacteria. Results showed that pathogen-host interactions were highly dynamically regulated, with corresponding fluctuations in host and pathogen transcriptomes during infection. The expression levels of many immunogenes involved in cytokine-cytokine receptor,Toll-like receptor signaling, and other immunerelated pathways were significantly up-regulated during the infection period. Furthermore, metabolic processes and the use of oxygen in L. crocea were strongly affected by P. plecoglossicida infection. The WGCNA results showed that the metabolic process was strongly related to the entire immune process.For P. plecoglossicida, the expression levels of motility-related genes and flagellum assemblyrelated genes were significantly up-regulated. The results of this study may help to elucidate the interactions between L. crocea and P.plecoglossicida.

关键词： larimichthys crocea Pseudomonas plecoglossicida Dual RNA-seq Host-pathogen interactions Dynamic transcriptome

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Antibacterial and antiparasitic activities analysis of a hepcidinlike antimicrobial peptide from larimichthys crocea

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Acta Oceanologica Sinica 2020年第10期39卷 129-139页

作者： Libing Zheng Yuan Li Jun Wang Ying Pan Jia Chen Weiqiang Zheng Longshan Lin State Key Laboratory of Large Yellow Croaker Breeding Ningde 352103China Third Institute of Oceanography Ministry of Natural ResourcesXiamen 361005China National and Provincial Joint Laboratory of Exploration and Utilization of Marine Aquatic Genetic Resources School of Marine Science and TechnologyZhejiang Ocean UniversityZhoushan 316022China

As an economically important marine fish,the large yellow croaker larimichthys crocea suffered from marine white spot disease caused by the ectoparasite Cryptocaryon irritans in recent years.This disease not only could result in physiological damage,but also lead to secondary bacterial invasion.Reports indicated some AMPs(antimicrobial peptides)were of antiparasitic activity to C.irritans.Hepcidin-like(Lc-HepL)was one of the significant differential expression genes excavated from the transcriptome following a challenge with C.irritans.In this study,we characterized this AMP’s bioactivity based on the levels of mRNA and protein.After challenged by C.irritans,qRT-PCR showed Lc-HepL was significantly upregulated in six tissues,including gill,muscle,liver,head kidney and spleen during theront infection,trophont falling off,and secondary bacterial invasion stages,which implicated a role Lc-HepL played in the immune defense against C.irritans and secondary bacterial infection.Recombinant Lc-HepL(rLc-HepL)was induced and purified successfully.rLc-HepL exhibited antibacterial activity to certain bacteria in a dose-and time-dependent manners.Anti-C.irritans activity was explored for the first time and found it could cause the theronts membrane rupture and contents leakage.These results provided the first evidence that Lc-HepL had strong antiparasitic activity against marine fish ectoparasites C.irritans theronts.Together,data indicated that Lc-HepL might be an important component in the innate immune system against C.irritans and has the potential to be employed in future drug development.

关键词： larimichthys crocea Cryptocaryon irritans hepcidin-like antibacterial activity antiparasitic activity

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Optimal amounts of coconut oil in diets improve the growth,antioxidant capacity and lipid metabolism of large yellow croaker(larimichthys crocea)

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Marine Life Science & Technology 2020年第4期2卷 376-385页

作者： Tao Ding Ning Xu Yongtao Liu Xueshan Li Xiaojun Xiang Dan Xu Chuanwei Yao Qiangde Liu Zhaoyang Yin Kangsen Mai Qinghui Ai Key Laboratory of Aquaculture Nutrition and Feed Ministry of Agriculture and Rural Affairsand Key Laboratory of MaricultureMinistry of EducationOcean University of China.Qingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237China

The purpose of this study was to investigate the effect of different dietary coconut oil(CO)levels on growth,antioxidant capacity and lipid metabolism of juvenile large yellow croaker(larimichthys crocea).Five iso-nitrogen(45%crude protein)and iso-lipid(13%crude lipid)experimental diets were prepared by replacing 0%(the control),25%,50%,75%and 100%fish oil with coconut oil.The results showed that dietary CO had no significant effect on survival rate(SR.P>0.05).However,the specific growth rate was increased significantly when compared with the control group when fish were fed the diet with 50%CO(P<0.05).The saturated fatty acids were increased significantly with increasing dietary CO in the liver and muscle,whereas the content of n-3 PUFA was decreased significantly(P<0.05).The highest activities of glutathione peroxidase and superoxide dismutase in the liver were recorded in fish-fed diet with 50%CO;conversely,the content of malondialdehyde was significantly decreased(P<0.05).The mRNA expression of peroxisome proliferator-activated receptor a,carnitine palmitoyl transferase 1 and acyl-CoA oxidase reached the highest levels in fish-fed diet with 50%CO.To some extent,this indicated that the rapid oxidation reaction of fatty acids to provide energy may be the reason for the rapid growth of large yellow croaker.In conclusion,fish-fed diet with 50%CO increased the growth rate and antioxidant capacity.Therefore,the optimal replacement level of CO to FO in the diet should be 50%.

关键词： Coconut oil Growth performance Antioxidant capacity Lipid metabolism larimichthys crocea

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Effect of fasting on protein metabolism in muscle tissue of larimichthys crocea revealed by transcriptome and proteome

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Journal of Oceanology and Limnology 2021年第5期39卷 1972-1984页

作者： Yu ZHANG Weiliang SHEN Juan LI Baoxiao ZHENG Zhengjia LOU Mohammad Aslam HOSAIN Baoying QIAN Liangyi XUE College of Marine Sciences Ningbo UniversityNingbo 315832China Ningbo Institute of Marine and Fisheries Ningbo 315103China School of Life Science Taizhou UniversityTaizhou 318000China

The large yellow croaker(larimichthys crocea)is an important mariculture fish in China.Farmed large yellow croaker undergo periods of fasting to adapt to the environment or to improve meat quality.To better understand the physiological responses of their muscle tissues to fasting stresses,we analyzed the transcriptomes and proteome s of both normally-fed and fasting fish groups and identified 7578 differentially expressed genes(DEGs)and 297 differentially expressed proteins(DEPs)among them.Gene ontology and KEGG analysis showed that the enriched biological pathways were mainly involved in various synthetic and catabolic pathways,especially the protein metabolism.Based on the omics data,nine DEGs related to muscle composition(CAN3,MYL3,and TNNC2),growth(MSTN and MYF5),autophagy(TSC2 and ULK1),and the ubiquitin proteasome pathway(PRS6 B and UCHL3)were examined using qPCR.In response to fasting stress,MYL3 and TNNC2 were significantly downregulated,while genes associated with autophagy and the ubiquitin proteasome pathway were significantly upregulated.In re sponse to fasting stres s,MYL3,TNNC2,and MYF5 positively correlated with muscle growth were significantly downregulated,while inhibiting growth MSTN and genes associated with autophagy and the ubiquitin proteasome pathways were significantly upregulated.These results clarify the effects of fasting on metabolic changes in their muscle components and growth at the molecular level.

关键词： larimichthys crocea fasting transcriptome proteome muscle

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Characterization of ghrelin mRNA expression in fasting larimichthys crocea juveniles

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Acta Oceanologica Sinica 2021年第6期40卷 35-40页

作者： Feng Liu Xiaokang Lv Tianqi Chu Mengjie Wang Wei Zhan Bao Lou Institute of Hydrobiology Zhejiang Academy of Agricultural SciencesHangzhou 310021China Key Laboratory of Mariculture and Enhancement of Zhejiang Province Marine Fisheries Research Institute of ZhejiangZhoushan 316021China School of Fishery Zhejiang Ocean UniversityZhoushan 316022China

larimichthys crocea is a marine fish species cultured in China.Short-term starvation is often applied to improve the quality of cultured L.crocea,and the expression of ghrelin in tissues of stomach,muscle,brain,intestines,liver,and kidney,involved in starvation response,under starvation conditions were studied to understand the effect of starvation on the expression of ghrelin in L.crocea juveniles.The ghrelin expression was tissue-specific,and expression was significantly higher in the stomach compared to other tissues(Pcrocea juveniles.This study provids insights into ghrelin function and an important reference for the development of reasonable feeding strategies for L.crocea juveniles.

关键词： larimichthys crocea starvation ghrelin gene expression

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Isolation and characterization of polymorphic microsatellite loci in large yellow croaker, larimichthys crocea

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Acta Oceanologica Sinica 2012年第4期31卷 149-153页

作者： YE Hua REN Peng ZHAO Guangtai YUE Genhua WANG Zhiyong Fisheries Breeding and Healthy Cultivation Research Centre Southwest UniversityChongqing402460China Key Laboratory of Healthy Mariculture for the East China Sea Ministry of Agriculture of the P.R.C.Jimei UniversityXiamen 361021China Molecular Population Genetics Group Temasek Life Sciences Laboratory117604 Singapore

An （AC）n-microsatellite-enriched library for larimichthys crocea was constructed in this study. Primers for fifty simple sequence repeat （SSR） loci were synthesized and genotyped on 30 L. crocea individuals from Guanjingyang wild population （WP） in Fujian Province and 38 individuals from Ningbo cultured population （CP） in Zhejiang Province. Only 21 loci were successfully amplified and polymorphic in two populations. In WP, the observed heterozygosity （Ho） ranged from 0.233 to 0.900 and the expected heterozygosity （HE） ranged from 0.326 to 0.893, with an average of 7.8 alleles/locus, the polymorphism information content （PIC） ranged from 0.283 to 0.866 （mean 0.731）. In CP, the Ho ranged from 0.189 to 0.892 and the HE ranged from 0.333 to 0.800, with an average of 4.4 alleles/locus. The probability test showed significant departures from HWE in 9 and 2 of the 21 loci in WP and in CP, respectively. Deficiency of heterozygotes at four loci showed the presence of null alleles （P 〈0.01）. The PIC of 20 microsatellite loci in WP were greater than 0.50. Overall, these novel markers are potentially useful for the study of population genetics, construction of genetic linkage and quantitative trait loci maps in large yellow croaker.

关键词： larimichthys crocea microsatellites microsatellite-enriched library genetic diversity

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Cloning,Functional Characterization and Expression Analysis of the elovl4a Gene in the Large Yellow Croaker(larimichthys crocea)

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Journal of Ocean University of China 2021年第6期20卷 1515-1527页

作者： ZHANG Meng LI Wanbo ZHU Yangyang WANG Weijia WANG Zhiyong Key Laboratory of Healthy Mariculture for the East China Sea Ministry of Agriculture and Rural AffairsJimei UniversityXiamen 361021China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266235China Fujian Engineering Research Center of Aquatic Breeding and Healthy Aquaculture Jimei UniversityXiamen 361021China

Elovl4 is a fatty acyl elongase which participates in the biosynthesis of long-chain polyunsaturated fatty acids(LCPUFA).Herein we isolated and functionally characterized the elovl4a gene in the large yellow croaker larimichthys crocea,and investigated the regulatory effects of transcription factors Hnf4α,Lxrα,Pparαon the activity of elovl4a promoter.Tissue expression pattern revealed that elovl4a was widely expressed in several tissues and predominantly in brain.Heterologous expression in yeast showed that L.crocea Elovl4a could effectively elongate both C18 and C20 PUFA substrates to C22 fatty acid.In addition,the affinity of large yellow croaker Elovl4a to n-6 series fatty acids is weak and cannot prolong 18:2n-6 and 18:3n-6.Moreover,L.crocea elovl4a reporter activities were elevated by 1.31-,1.39-and 1.48-fold via over-expression of Lxrα,Pparαand Hnf4α,respectively.The above findings can enrich the knowledge of biosynthesis pathway of LC-PUFA in the large yellow croaker and contribute to elucidate the LC-PUFA anabolism mechanism in fish.

关键词： larimichthys crocea elovl4a long-chain polyunsaturated fatty acids promoter

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