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1.
J Fish Biol ; 104(2): 463-472, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36808734

RESUMO

Brachymystax tsinlingensis Li is a threatened fish species endemic to China. With the problems of environmental factors and seeding breeding diseases, it is important to further improve the efficiency of seeding breeding and the basis of resource protection. This study investigated the acute toxicity of copper, zinc and methylene blue (MB) on hatching, survival, morphology, heart rate (HR) and stress behaviour of B. tsinlingensis. Eggs (diameter: 3.86 ± 0.07 mm, weight: 0.032 ± 0.004 g) of B. tsinlingensis were selected randomly from artificial propagation and developed from eye-pigmentation-stage embryos to yolk-sac stage larvae (length: 12.40 ± 0.02 mm, weight: 0.03 ± 0.001 g) and exposed to different concentrations of Cu, Zn and MB for 144 h in a series of semi-static toxicity tests. The acute toxicity tests indicated that the 96-h median lethal concentration (LC50 ) values of the embryos and larvae were 1.71 and 0.22 mg l-1 for copper and 2.57 and 2.72 mg l-1 for zinc, respectively, whereas the MB LC50 after 144-h exposure for embryos and larvae were 67.88 and 17.81 mg l-1 , respectively. The safe concentrations of copper, zinc and MB were 0.17, 0.77 and 6.79 mg l-1 for embryos and 0.03, 0.03 and 1.78 mg l-1 for larvae, respectively. Copper, zinc and MB treatments with concentrations greater than 1.60, 2.00 and 60.00 mg l-1 , respectively, led to a significantly low hatching rate and significantly high embryo mortality (P < 0.05), and copper and MB treatments with concentrations greater than 0.2 and 20 mg l-1 led to significantly high larvae mortality (P < 0.05). Exposure to copper, zinc and MB resulted in developmental defects, including spinal curvature, tail deformity, vascular system anomalies and discolouration. Moreover, copper exposure significantly reduced the HR of larvae (P < 0.05). The embryos exhibited an obvious change in behaviour, converting from the normal behaviour of emerging from the membrane head first to emerging tail first, with probabilities of 34.82%, 14.81% and 49.07% under copper, zinc and MB treatments, respectively. The results demonstrated that the sensitivity of yolk-sac larvae to copper and MB was significantly higher than that of embryos (P < 0.05) and that B. tsinlingensis embryos or larvae might be more resistant to copper, zinc and MB than other members of the Salmonidae family, which benefits their resource protection and restoration.


Assuntos
Salmonidae , Poluentes Químicos da Água , Animais , Cobre/toxicidade , Larva , Zinco/toxicidade , Aquicultura , Poluentes Químicos da Água/toxicidade , Embrião não Mamífero
2.
Pestic Biochem Physiol ; 196: 105607, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37945248

RESUMO

Due to chemical and photochemical stability, triazophos has been frequently detected in rivers and oceans over the years with extensive use for pest control in agriculture, and it has become a worldwide ecological concern to the aquatic environment. Until now, fewer data are available regarding the potential long-term adverse effects of triazophos on aquatic invertebrates, which plays an essential role in aquatic food webs, as a key group for water ecosystems. In this experiment, the F1- and F2 progenies of Daphnia magna were recovered when daphnias (F0) exposure to triazophos at environmental-related concentrations (0.1 and 1.0 µg/L) for 21 d; and the indexes related to phenotypic traits, reproduction and gene expression were measured in tested animals. The results showed that heart rate and total number of neonates in exposed F0-daphnias were significantly lower than those of control group, and the detoxification genes (HR96 and P-gp) were up-regulated while genes related reproduction (Vtg) and molting (Nvd and Shd) were significantly down-regulated. The heart rate and individual size of F1-daphnias (<24 h) were significantly reduced in the treatment group. After 21-d recovery, the heart rate and expression of HR96, P-gp, Vtg, Nvd and Shd were declined in F1-daphnias. There was no obvious difference of morphological traits and heart rate between treatment and control in F2-daphnias (<24 h). In summary, daphnias (F0) exposure to triazophos with environmental dose could raise toxic effects on its offspring (F1), which is mainly manifested by reduced heart rate, the accumulated number and individual size of offspring and decreased expression of genes related to molting and reproduction.


Assuntos
Daphnia , Poluentes Químicos da Água , Animais , Daphnia/genética , Ecossistema , Poluentes Químicos da Água/toxicidade , Poluentes Químicos da Água/metabolismo , Reprodução
3.
Fish Physiol Biochem ; 49(1): 155-167, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36547499

RESUMO

The melanocortin-3-receptor (MC3R) plays an important role in mammals' food intake and energy homeostasis. However, its physiological role in bony fishes, such as grass carp, has not been well understood. This study reports the molecular cloning, tissue distribution, and pharmacological characterization of grass carp melanocortin-3-receptor (ciMC3R). Phylogenetic and chromosomal synteny analyses indicated that ciMC3R was closest to cyprinid fishes in evolution. Quantitative PCR experiments revealed that the mRNA of ciMC3R was highly expressed in the brain of grass carp. The cytological function of ciMC3R was investigated by the co-transfection of pcDNA3.1-ciMC3R and the signal-pathway-specific luciferase into the HEK293T cells. Results revealed that the four agonists, α-MSH, ß-MSH, ACTH, and NDP-MSH, potentiate the activation of ciMC3R and further increase the production of cAMP and upregulate the MAPK/ERK signaling, respectively. Our study will provide basic data for exploring the physiological functions of grass carp MC3R, especially in energy homeostasis and food intake.


Assuntos
Carpas , Proteínas de Peixes , Receptor Tipo 3 de Melanocortina , Animais , Humanos , Carpas/genética , Clonagem Molecular , Proteínas de Peixes/genética , Células HEK293 , Filogenia , Receptor Tipo 3 de Melanocortina/genética
4.
Fish Physiol Biochem ; 49(1): 61-74, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36525144

RESUMO

Melanocortin 3 and 4 receptors are two important neural G protein-coupled receptors that regulate energy homeostasis in vertebrates. Melanocortin receptor accessory protein 2 (MRAP2) is also involved in the regulation of food intake and body weight as a variable regulator of melanocortin receptors. Rainbow trout (Oncorhynchus mykiss) is a valuable cold-water fish cultured worldwide. In the rainbow trout model, we cloned and identified mrap2a, a paralog of mrap2. Rainbow trout mrap2a consisted of a 690 bp ORF and was expected to encode a putative protein of 229 amino acids. The qPCR results showed that rainbow trout mrap2a was expressed at high levels in brain tissue similar to mc3r and mc4r. In addition, co-immunoprecipitation verified that MRAP2a interacts with MC3R and MC4R in vitro and that MRAP2a is involved in and regulates the constitutive activity and signaling of MC3R and MC4R. MRAP2a reduced constitutive and agonist-stimulated cAMP levels of MC3R; furthermore, MRAP2a increased constitutive ERK1/2 activation but reduced ligand-induced stimulation at high levels of expression. For MC4R, MRAP2a showed decreased cAMP basal activity but increased agonist-stimulated cAMP signaling and increased ACTH ligand sensitivity. However, MRAP2a failed to affect MC4R constitutive activity and agonist-induced ERK1/2 signaling. Undoubtedly, our study will have great significance for revealing the conserved role of MC4R and MC3R signaling in teleost fish, especially in cold-water fish growth and energy homeostasis.


Assuntos
Oncorhynchus mykiss , Animais , Oncorhynchus mykiss/genética , Ligantes , Receptores de Melanocortina , Transdução de Sinais , Peso Corporal
5.
J Nanobiotechnology ; 20(1): 306, 2022 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-35761380

RESUMO

Protein-stabilized gold nanoclusters (Prot-Au NCs) have been widely used in biosensing and cell imaging owing to their excellent optical properties and low biotoxicity. However, several Prot-Au NCs reported in the literature do not retain the biological role of the protein, which greatly limits their ability to directly detect biomarkers. This study demonstrated for the first time the successful synthesis of dual-function avidin-stabilized gold nanoclusters (Av-Au NCs) using a one-pot method. The resulting Av-Au NCs exhibited intense blue and red emissions under 374 nm excitation. Furthermore, the Av-Au NCs retained the native functionality of avidin to bind to biotin. When DNA strands modified with biotin at both ends (i.e., linker chains) were mixed with Av-Au NCs, large polymers were formed, indicating that Av-Au NCs could achieve fluorescence signal amplification by interacting with biotin. Taking advantage of the aforementioned properties, we constructed a novel enzyme-free fluorescent biosensor based on the Av-Au NCs-biotin system to detect DNA. The designed fluorescent biosensor could detect target DNA down to 0.043 nM, with a wide line range from 0.2 nM to 20 µM. Thus, these dual-functional Av-Au NCs were shown to be an excellent fluorescent material for biosensing.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Avidina , Técnicas Biossensoriais/métodos , Biotina , Corantes , Ouro
6.
Pestic Biochem Physiol ; 182: 105036, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35249646

RESUMO

Residues of triazophos in aquatic ecosystems due to extensive use for controlling pests in agriculture has became worldwide concern, while the toxic response of triazophos on the non-target green algae in aquatic environment is not well studied. Therefore, the acute (96 h) toxic effects of 1 and 10 mg/L triazophos on green algae Chlorella pyrenoidosa were evaluated in present study. The results showed that the growth was notably inhibited when treated with triazophos and the 96 h-EC50 (median inhibition concentration) were 12.79 mg/L. The content of photosynthetic pigments (including chl a, chl b, total-chl and carotinoids) clearly decreased under two treatments after 48 h and 96 h with exception for the values at 48 h exposure in 1 mg/L treatment. In addition, the transcript abundance of photosynthesis-related genes (psbA, psbC and rbcL) showed obvious decrease in above two treatments after exposure 96 h to triazophos. In response to 10 mg/L triazophos treatment, the morphology of thylakoid chloroplast of algal cells were obviously damaged. It was also found that starch granules increased with down-regulation of atpB gene expression in 10 mg/L treatment, which suggests that triazophos may inhibit the energy metabolism of C. pyrenoidosa. Moreover, the algal growth inhibition was along with the increase of intracellular reactive oxygen species (ROS), activity of antioxidant enzymes and malondialdehyde content indicating oxidative damage and lipid peroxidation in the algal cells. Our findings reveal that triazophos has potential toxicity and environmental risks to one of the primary producers green algae.


Assuntos
Chlorella , Poluentes Químicos da Água , Chlorella/metabolismo , Ecossistema , Organotiofosfatos/toxicidade , Triazóis/farmacologia , Poluentes Químicos da Água/toxicidade
9.
Oncol Lett ; 21(4): 316, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33692848

RESUMO

Long intergenic non-protein coding RNA 52 (LINC00052) is a non-coding RNA with >200 nucleotides in length, which exerts important effects on several physiological and pathological processes of the human body. Recent studies have demonstrated that LINC00052 plays key roles in the tumorigenesis, progression and metastasis of multiple types of human cancer, including hepatocellular carcinoma, breast cancer, colorectal cancer, cervical carcinoma and gastric cancer. However, the associations between LINC00052 and these tumors remain unclear. The present review summarizes the biological functions of LINC00052 during the pathogenic process of certain tumors, and discusses its potential therapeutic targets.

10.
Biosens Bioelectron ; 179: 113059, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33561664

RESUMO

Herein, we report a novel dual-quenching electrochemiluminescence (ECL) immunosensor for detecting protein induced by vitamin K absence or antagonist-II (PIVKA-II) based on ECL resonance energy transfer (ECL-RET). In this protocol, self-accelerated ECL hybrids of CeO2 and Au nanoparticles functionalized g-C3N4 nanosheets (CeO2-Au-g-C3N4) were prepared, which exhibited high ECL emission in the presence of S2O82- as a coreactant for "signal on" state. Concretely, CeO2 with a reproducible redox couple of Ce3+ and Ce4+ could act as an efficient co-reaction accelerator to generate more oxidizing intermediate (SO4•-) to significantly self-promote the ECL emission of g-C3N4 NSs/S2O82- ECL system. Besides, Au nanoparticles not only accelerated electron transfer in the ECL process, but also provided massive active sites for biomolecules immobilization. The dual quenching labels of Ag nanocubes modified with vitamin B2 (AgNCs-VB2) were firstly proposed towards g-C3N4 NSs/S2O82- ECL system by ECL-RET, resulting in the remarkable ECL decrease for "signal off" state. Based on the sandwich immunoreaction, the "on-off" PIVKA-II ECL immunosensor gratifyingly possessed excellent detection sensitivity with the linear range of 0.4 pg mL-1-10 ng mL-1 and the low detection limit of 28.46 fg mL-1 (S/N = 3). This presented strategy might provide a potential alternative tool for PIVKA-II detection in medical research and early clinical diagnostics of hepatocellular carcinoma.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Técnicas Eletroquímicas , Ouro , Imunoensaio , Limite de Detecção , Medições Luminescentes , Prata
11.
Artigo em Inglês | MEDLINE | ID: mdl-31654829

RESUMO

Extensive use of triazophos for the chemical control of pests in agriculture or aquaculture might strongly disturb the aquatic environment due to residue accumulation through various routes like surface run-off, spray-drift and effluent from factories, which have potential negative effects to non-target aquatic organisms. Previous studies have documented the antioxidative effects of triazophos to mammals, however, the oxidative toxicity of triazophos to fish has not been adequately studied to date. Thus, an acute exposure (96 h) to triazophos at different concentrations of 0.06, 0.3 and 1.5 mg/L (corresponding to 1/50th, 1/10th and 1/2th of 96 h-LC50, respectively), was conducted to investigate the triazophos-induced oxidative stress in adult zebrafish (Danio rerio). The results showed that the time- and dose-dependent induction of oxidative stress except for the ROS level in liver after 24 and 48 h exposure, as indicated by increased reactive oxygen species (ROS), malondialdehyde (MDA) level and a compromised antioxidant defense system, including increased superoxide dismutase (SOD) activity, catalase (CAT) activity, glutathione (GSH) content as well as the increased at first and decreased afterwards genes expression (Sod1, Sod2, Cat and Gpx) in brain. Simultaneously, ROS and MDA showed an increased trend, SOD activity, CAT activity and GSH content showed a trend of increasing at 24 h and decreasing at 48 h, and then increasing at 96 h and Sod1, Sod2, Cat and Gpx gene showed decreasing at first and then increasing in liver tissue. The present study concluded that the damage of the antioxidant system by triazophos induced oxidative stress in the brain and liver of zebrafish with concomitant lipid peroxidation, which is an important mechanism underlying the triazophos-induced acute toxicity.


Assuntos
Encéfalo/metabolismo , Fígado/metabolismo , Organotiofosfatos/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Triazóis/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo , Animais , Catalase/metabolismo , Glutationa/metabolismo , Peroxidação de Lipídeos , Malondialdeído/metabolismo , Estresse Oxidativo , Superóxido Dismutase/metabolismo
12.
Pestic Biochem Physiol ; 158: 135-142, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31378349

RESUMO

Triflumizole is one of imidazole fungicides that works by inhibiting ergosterol biosynthesis, and is widely used for the control of powdery mildew and scabs on various fruits and crops. Triflumizole residue has been frequently detected in soil and aquatic ecosystems. While many studies have focused on its toxic effect on terrestrial and aquatic animals, little attention has been paid to aquatic algae, the primary producers of aquatic environments. Therefore, we evaluated the acute (96 h) toxicity effects of triflumizole on the freshwater algae Chlorella vulgaris, by examining growth, cell morphology, photosynthesis, and oxidative stress. The results showed that the 96 h median inhibition concentration (96 h-EC50) was 0.82 mg/L (95% confidential interval 0.70-0.98 mg/L).The growth of algal cells was conspicuously inhibited by triflumizole exposure, and the cell surfaces appeared to be shrunkThe chlorophyll content (including Chl-a, Chl-b and T-Chl) dramatically decreased at triflumizole concentrations of 0.2 and 1.0 mg/L. In addition, the transcript abundance of photosynthesis-related genes (psaB, psbC and rbcL) showed obvious decreases in above treatments after 96 h of exposure to triflumizole. Moreover, the algal growth inhibition was accompanied by an increase in intracellular reactive oxygen species and malondialdehyde content, as well as increased activity of antioxidant enzymes such as superoxide dismutase and peroxidase, indicating oxidative stress and lipid peroxidation. Our findings reveal that triflumizole has potential toxicity to the primary producers (freshwater algae) in aquatic ecosystems.


Assuntos
Chlorella vulgaris/efeitos dos fármacos , Imidazóis/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidase/metabolismo , Fotossíntese/efeitos dos fármacos , Superóxido Dismutase/metabolismo
13.
Mitochondrial DNA A DNA Mapp Seq Anal ; 30(5): 695-701, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31286805

RESUMO

Brachymystax tsinlingensis Li, distributed scatteredly in Qinling Mountains of China, is an ideal material for studies of conservation and phylogeography. In the present study, the genetic variations and phylogeographical patterns of B. tsinlingensis collecting from the Yangtze River (n = 30) and the Yellow River (n = 70) were compared based on 1082-1163 bp control region with partial tRNA and 912 bp cyt b partial sequence of mtDNA. Analysis of variance indicated that a high proportion of the total genetic variance (96.03%) was distributed among populations, supporting strong geographic structuring of mtDNA polymorphism. Ten haplotypes were assigned to two clades that were related to geographic regions. No haplotype shared between geographic populations from the Yangtze River and the Yellow River, and two clades related to geographic regions can be clearly found from phylogenetic analysis by Bayesian approach, one from individuals in the Yangtze River and the other from individuals in the Yellow River, which probably indicated artificial carrying not happened historically. The mitochondrial results revealed an obvious differentiation of genetic structure of B. tsinlingensis between populations of Yellow River and Yangtze River, and the estimated diverge time around 0.20 MY. It was suggested that each of the two evolutionarily distinct groups of B. tsinlingensis should be protected and crossing individuals between two river systems should be avoided for conservation purpose.


Assuntos
Extinção Biológica , Genoma Mitocondrial/genética , Rios , Salmonidae/classificação , Salmonidae/genética , Distribuição Animal , Animais , China , Código de Barras de DNA Taxonômico , Especificidade da Espécie
14.
J Cell Biochem ; 120(10): 17258-17272, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31104316

RESUMO

The dysregulation of long-chain noncoding ribonucleic acid (lncRNA) is a common phenomenon in many human cancers. Some studies on the biological function of long intergenic non-protein-coding RNA 52 (LINC00052) in cancer indicate that this gene can act as either oncogene or tumor suppressor in some kinds of cancers, such as breast cancer, gastric cancer, liver cancer, and lung cancer. However, the biological function of LINC00052 in colorectal cancer (CRC) has not been studied. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot (WB) techniques were applied to detect the expression levels of LINC00052, miR-574-5p, and calcium-binding and coiled-coil domain 1 (CALCOCO1) in CRC cells and tissues. We authenticated the biological function of LINC00052 and miR-574-5p in CRC, and find some target genes for LINC00052 and miR-574-5p via bioinformatics methods. Dual-luciferase reporter gene assay was performed to identify the interaction between LINC00052 and miR-574-5p or CALCOCO1 and miR-574-5p. The results demonstrated that LINC00052 was downregulated in CRC tissues compared with their adjacent tissues. And LINC00052 could suppress CRC cells metastasis both in vivo and in vitro. Beyond that, miR-574-5p was upregulated in CRC tissues, and as an oncogene, it accelerated CRC cell migration and invasion. More importantly, the results of our research demonstrated that LINC00052 could regulate the expression of CALCOCO1 via sponging miR-574-5p in CRC. Overall, our study illuminated the lncRNA-miRNA functional networks in CRC, and these results might provide a new research direction for the diagnosis and treatment of CRC.


Assuntos
Biomarcadores Tumorais/metabolismo , Movimento Celular , Neoplasias Colorretais/prevenção & controle , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/prevenção & controle , MicroRNAs/antagonistas & inibidores , RNA Longo não Codificante/genética , Animais , Apoptose , Biomarcadores Tumorais/genética , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Ciclo Celular , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundário , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , MicroRNAs/metabolismo , Invasividade Neoplásica , Prognóstico , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
15.
Ecotoxicol Environ Saf ; 180: 646-655, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31136875

RESUMO

Triazophos (TAP) has become a part of widespread pollutant of the aquatic environment due to its residue. Current study was designed to investigate the toxic effect of TAP at different doses (0.06, 0.3 and 1.5 mg/L) to the model organism of zebrafish (Danio rerio) by using multi-endpoint analysis in a 96 h acute exposure test. The direct observation that histological and ultrastructural alteration of zebrafish brain and liver were carried out via paraffin section in hematoxylin and eosin (H&E) staining and transmission electron microscopy (TEM), respectively. In addition, a series of methods were applied for exploring the physiological parameters related to cellular apoptosis. Results indicated that vacuolar structure after 96 h treatment with TAP were appeared in the molecular and granular layers of cerebellum. A large number of nuclear retraction, tissues vacuolation and cytoplasmic loss were observed in liver at histological level. From the fine structural level, the mitochondrial vacuolation and membrane damage of brain cells were found and the cristae of mitochondria disintegrated partly in hepatocytes. Onset of such histological structure alterations were one of the most intuitive reflection to TAP exposure, which needs to analyze biochemical alterations for further study. The mitochondrial membrane potential (MMP) showed a downward trend in the brain and liver of zebrafish. Simultaneously, the activity of caspase-3 and caspase-9 increased after 96 h exposure with a concentration-dependent manner, which could be served as a suitable indicator of cellular apoptosis. Furthermore, apoptosis-related genes (Apaf-1, p53, Bax, Bcl-2, caspase-3 and caspase-9) transcription showed different alterations in response to the TAP treatment. These results indicated that TAP exposure led to apoptosis in zebrafish brain and liver and it was speculated that the apoptosis may occur through mitochondrial pathway. The present study demonstrated that the exposure of zebrafish to the insecticide TAP led to observe its effects at both histological structure and apoptosis level in liver and brain.


Assuntos
Apoptose/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Exposição Ambiental/efeitos adversos , Fígado/efeitos dos fármacos , Organotiofosfatos/toxicidade , Triazóis/toxicidade , Animais , Encéfalo/patologia , Fígado/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Peixe-Zebra , Proteínas de Peixe-Zebra/metabolismo
16.
Fish Shellfish Immunol ; 86: 882-891, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30553892

RESUMO

Ferritin is a protein related to the storage of iron and widely distributed in animals. It participates in many biological process, including antioxidation, cell activation, angiogenesis, regulation of iron metabolic balance and immune defense. In the present study, a novel ferritin gene was identified from red swamp crayfish Procambarus clarkii, with a cDNA sequence encoding a predicted 221 amino-acid residues. The ferritin protein contains a 19-residue signal peptide and 145-residue classic ferritin domain. The NJ phylogenetic analysis showed PcFer clustered with other crustacean peptides. The recombinant PcFer protein was produced and purified in E. coli, and the anti-rabbit polyclonal antibody was obtained. The rPcFer exhibited iron binding activity at a dose-dependent effect. The qPCR and western blot analysis revealed that PcFer was highly expressed in hemocytes, hepatopancreas, and gills. After challenged with WSSV and Aeromonas hydrophila, the mRNA and protein expression patterns of PcFer were significantly up-regulated in hemocytes and hepatopancreas. dsRNA interfering technique was utilized to silence the expression of PcFer gene. The WSSV copy number in PcFer silenced shrimp was much higher than that in the control group. The present study indicated that PcFer was involved in the immune defense against WSSV and Aeromonas hydrophila, and might inhibit WSSV replication in P. clarkii. These results will provide important data support for further study of the functional role of the ferritin gene.


Assuntos
Astacoidea/genética , Astacoidea/imunologia , Ferritinas/genética , Ferritinas/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Ferritinas/química , Perfilação da Expressão Gênica , Brânquias/metabolismo , Hemócitos/metabolismo , Hepatopâncreas/metabolismo , Filogenia , Replicação Viral , Vírus da Síndrome da Mancha Branca 1/fisiologia
17.
Oncotarget ; 8(16): 27105-27119, 2017 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-28404920

RESUMO

The liver provides vital metabolic, exocrine and endocrine functions in the body as such pathological conditions of the liver lead to high morbidity and mortality. The liver is highly regenerative and contains facultative stem cells that become activated during injury to replicate to fully recover mass and function. Canonical Wnt/ß-catenin signaling plays an important role in regulating the proliferation and differentiation of liver progenitor cells during liver regeneration. However, possible roles of noncanonical Wnts in liver development and regeneration remain undefined. We previously established a reversibly-immortalized hepatic progenitor cell line (iHPx), which retains hepatic differentiation potential. Here, we analyze the expression pattern of the essential components of both canonical and noncanonical Wnt signaling pathways at different postnatal stages of mouse liver tissues and iHPx cells. We find that noncanonical Wnt4, Wnt5a, Wnt9b, Wnt10a and Wnt10b, are highly expressed concordantly with the high levels of canonical Wnts in late stages of liver tissues. Wnt5a, Wnt9b, Wnt10a and Wnt10b are able to antagonize Wnt3a-induced ß-catenin/TCF activity, reduce the stemness of iHPx cells, and promote hepatic differentiation of liver progenitors. Stem cell implantation assay demonstrates that Wnt5a, Wnt9b, Wnt10a and Wnt10b can inhibit cell proliferation and promote hepatic differentiation of the iHPx progenitor cells. Our results strongly suggest that noncanonical Wnts may play an important role in fine-tuning Wnt/ß-catenin functions during liver development and liver regeneration. Thus, understanding regulatory mechanisms governing proliferation and differentiation of liver progenitor cells may hold great promise to facilitate liver regeneration and/or progenitor cell-based therapies for liver diseases.


Assuntos
Diferenciação Celular , Autorrenovação Celular , Fígado/citologia , Células-Tronco/citologia , Células-Tronco/metabolismo , Proteínas Wnt/metabolismo , Via de Sinalização Wnt , Animais , Biomarcadores , Diferenciação Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Autorrenovação Celular/genética , Expressão Gênica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Genes Reporter , Humanos , Fígado/fisiologia , Regeneração Hepática/genética , Masculino , Camundongos , Transplante de Células-Tronco , Transcriptoma
18.
Dev Comp Immunol ; 72: 69-78, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28238880

RESUMO

Lysozymes as an important immune factor, play vital roles in innate immune response against pathogen infection. In the present study, one c-type and g-type lysozymes were identified from Chinese giant salamander (Andrias davidianus). They shared highly conserved structural features with lysozymes from other species. Spatial expression analysis revealed that AdlysC transcript was most abundant in liver and stomach, and least in muscle and brain. In contrast, the expression level of AdlysG was most abundant in liver and least in muscle and skin. The transcription level of c-type and g-type lysozymes were up-regulated after Aeromonas hydrophila infection in liver and spleen, indicating their participations in the immune response. Moreover, the recombinant AdlysC and AdlysG protein were produced and purified, and were used to investigate the lysozyme activity at different pH and temperatures. The optimal lytic activity was determined at pH 6.0 and at a temperature of 30 °C. Through the minimal inhibitory concentration test, the rAdlysC and rAdlysG exhibited apparent antibacterial activity against both Gram-positive and Gram-negative bacteria with a variable concentration. In conclusion, it is the first report of lysozymes in A. davidianus, and c-type and g-type lysozymes should be involved in the innate immune response of A. davidianus.


Assuntos
Aeromonas hydrophila/imunologia , Anti-Infecciosos/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Fígado/metabolismo , Muramidase/metabolismo , Urodelos/imunologia , Animais , Bacteriólise , Galinhas , Gansos , Concentração de Íons de Hidrogênio , Imunidade Inata , Fígado/imunologia , Muramidase/genética , Isoformas de Proteínas/genética , Temperatura , Transcriptoma
19.
Dev Comp Immunol ; 70: 59-68, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28065604

RESUMO

Galectins are considered as a multifunctional protein which play essential roles in cell adhesion and apoptosis, inflammation, tumor progression and immune response. In spite of extensive studies of galectin importance in immune system among different animals, few studies have been devoted to their functions in amphibian. In the present study, we characterized one proto type of galectin (named AdGal1) from Chinese giant salamander Andrias davidianus and studied its function in immune response. AdGal1 cDNA possesses an open reading frame of 598 bp, which encodes a putative galectin of 134 amino acids containing one carbohydrate recognition domains (CRDs). The constitutive expression of mRNA transcripts was detected in a wide range of tissues, with the highest expression in kidney. Immune challenges with Aeromonas hydrophila and Chinese giant salamander iridovirus (GSIV), the transcript level of AdGal1 in kidney was significantly upregulated. The mature protein of AdGal1 was successfully expressed and purified in Escherichia coli BL21 (DE3). The recombinant AdGal1 (rAdGal1) could show bind activity to different Gram negative and Gram positive bacteria. It could also strongly agglutinate different kinds of bacteria at different concentrations. Collectively, these data from the present study indicate that AdGal1 is a vital pattern recognition receptor to recognize different microbes in the innate immune system of Andrias davidianus.


Assuntos
Aeromonas hydrophila/imunologia , Proteínas de Anfíbios/metabolismo , Infecções por Vírus de DNA/imunologia , Galectina 1/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Iridovirus/imunologia , Rim/imunologia , Urodelos/imunologia , Aglutinação , Proteínas de Anfíbios/genética , Animais , Aderência Bacteriana , Células Cultivadas , Clonagem Molecular , Galectina 1/genética , Imunidade Inata , Rim/microbiologia , Rim/virologia , Ligação Proteica , Transcriptoma , Regulação para Cima , Urodelos/virologia
20.
Oncotarget ; 7(30): 47593-47608, 2016 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-27351280

RESUMO

Long non-coding RNAs (LncRNAs) are a group of RNAs that are more than 200 nt in length but cannot encode proteins. Accumulating evidences showed that abnormal LncRNA expressions are highly involved in many kinds of tumor. By using gene trap methods which could knockdown gene expression to find important genes, we found one LncRNA which called intergenic non-protein coding RNA 52 (LINC00052) has the ability to inhibit invasion and migration of hepatocarcinoma cells. We found that invasion, migration and proliferation abilities in SMMC7721 cell were inhibited after up-expressing LINC00052. We identified that NTRK3 was the target gene of LINC00052. Down-expression of NTRK3 could increase SMMC7721 cell invasion, migration and proliferation. Meanwhile, we discovered that LINC00052 could regulate NTRK3 expression by forming complementary base pairing with miR-128 and miR-485-3p to reduce the luciferase activity of NTRK3 3'UTR. These results reveal a new mechanism for understanding hepatocarcinoma cells invasion and migration.


Assuntos
Carcinoma Hepatocelular/patologia , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/patologia , MicroRNAs/fisiologia , RNA Longo não Codificante/fisiologia , Receptor trkC/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Humanos , Invasividade Neoplásica
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