INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2021
·Vol. 22
·期 16
·8949
·
IF 5.924
Huiqing Luo,Na Li,Liyan Liu,Huaqiao Wang,Feng He
Fluorophores with aggregation-induced emission enhancement (AIEE) properties have attracted increasing interest in recent years. On the basis of our previous research, we successfully designed and synthesized eleven chalcones. Through an optical performance experiment, we confirmed that compounds1–6had obvious AIEE properties. As these AIEE molecules had excellent fluorescence properties and a large Stokes shift, we studied their application in living cell imaging, and the results showed that these compounds had low cytotoxicity and good biocompatibility at the experimental concentrations. More importantly, they could specifically label mitochondria. Subsequently, we selected zebrafish as experimental animals to explore the possibilities of these compounds in animal imaging. The fluorescence imaging of zebrafish showed that these AIEE molecules can enter the embryo and can be targeted to aggregate in the digestive tract, which provides a strong foundation for their practical application in the field of biological imaging. Compared with traditional fluorophores, these AIEE molecules have the advantages of possessing a small molecular weight and high flexibility. Therefore, they have excellent application prospects in the field of biological imaging. In addition, the findings of this study have very positive practical significance for the discovery of more AIEE molecules.Keywords:AIEE;chalcone;mitochondria;zebrafish
研究领域:
生物成像
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分子生物学
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材料科学
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产品关联
DOI
PubMed
高影响因子
MEDICAL SCIENCE MONITOR
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2018
·
IF 1.894
Gang Wang,Qi Xiao,Wenxiu Wu,Yao Wu,Yingjie Wei,Yue Jing,Zhunan Gong
Background Asiatic acid (AA; 2α,3β,23-trihydroxyurs-12-ene-28-oic acid) is an active compound derived from Centella asiatica, a traditional medicinal plant used widely in many Asian countries, particularly for the treatment of cancer. However, the modified AA derivative N-(2α,3β,23-acetoxyurs-12-en-28-oyl)-l-proline methyl ester (AA-PMe) has shown markedly better anti-tumor activity than AA. Material/Methods We evaluated the toxicity of AA and AA-PMe on zebrafish morphology, mortality, and hatching rate and determined the effect on SGC7901 cancer cells by acute toxicity assay. AA-PMe absorption in vitro in SGC7901 cells and in vivo in zebrafish was determined by establishing a highly accurate and reproducible HPLC protocol. Results In zebrafish, the toxicity of AA-PMe was lower than AA, with an acute toxic dose of AA-PMe above 25 μM, compared to acute toxicity at doses above 10 μM for AA. However, chronic toxicity of AA-PMe began occurring at doses below 25 μM but became apparent for AA at doses below 10 μM. Although low doses of AA-PMe were tolerated acutely, it became chronically toxic during zebrafish development, resulting in morphological abnormalities, including peripheral and abdominal edema, hemorrhage, abnormal body shape, enlarged yolk sac, and reduced motility. At low concentrations, absorption of AA-PMe by cells and zebrafish embryos occurred in a dose-dependent manner, but this stabilized as the concentration increased. Conclusions This pharmacokinetic study outlines the cellular and organismal effects of AA-PMe and suggests a theoretical basis that may underlie its mechanism of action.
研究领域:
药理学
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毒理学
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肿瘤学
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DOI
PubMed
高影响因子
Life-Basel
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2022
·Vol. 12
·期 1
·128
·
IF 3.251
Davide Di Paola,Sabrina Natale,Enrico Gugliandolo,Marika Cordaro,Rosalia Crupi,Rosalba Siracusa,Ramona D’Amico,Roberta Fusco,Daniela Impellizzeri,Salvatore Cuzzocrea,Nunziacarla Spanò,Fabio Marino,Alessio Filippo Peritore
Lipopolysaccharide (LPS), or bacterial endotoxin, is an important virulence factor in several human and animal pathologies. Oxazoline of Palmitoylethanolamide (PEAOXA) has shown strong anti-inflammatory activity in several animal models. LPS was applied for 24 h to zebrafish embryos to induce inflammation, and then the anti-inflammatory action of PEAOXA was evaluated for the first time in the zebrafish model (Danio rerio). Different concentrations of PEAOXA were tested for toxicity on zebrafish embryonic development; only the highest concentration of 30 mg/L showed toxic effects. Quantitative RT-PCR was applied to detect Tumor necrosis factor-α, Interleukin 1β, 6, and 8, and members of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB). Exposure to LPS induced an increase in pro-inflammatory cytokines (tumor necrosis factor and interleukin 1, 6, and 8) in both gene and protein expression, as well as an increase of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and the nuclear factor kappa light polypeptide enhancer in B-cells inhibitor (IκBα) gene expression. Furthermore, acute LPS exposure also induced an increase in tryptase release, related to mast cell activity, and in the production of apoptosis-related proteins (caspase 3, bax, and bcl-2). Treatment with PEAOXA 10 mg/L significantly counteracts LPS-induced inflammation in terms of cytokine expression and decreases tryptase release and the apoptosis pathway.Keywords:zebrafish;inflammation;PEAOXA;cytokines;apoptosis
研究领域:
免疫学
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发育生物学
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药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biology-Basel
·
2022
·Vol. 11
·期 8
·1243
·
IF 5.168
Michael Edbert Suryanto,Ferry Saputra,Kevin Adi Kurnia,Ross D. Vasquez,Marri Jmelou M. Roldan,Kelvin H.-C. Chen,Jong-Chin Huang,Chung-Der Hsiao
Simple SummaryWith the advancement of existing technology, artificial intelligence is widely applied in various fields of research, including cardiovascular studies. In this study, we explored the feasibility of conducting a markerless cardiac physiology assessment in zebrafish embryos by using DeepLabCut (DLC), a deep learning tool for motion analysis. Several cardiac parameters, such as heart rate, diastolic–systolic volumes (EDV/ESV), stroke volume, cardiac output, shortening fraction, and ejection fraction were obtained by the DLC-trained model and then compared to the previous published methods, Time Series Analysis and Kymograph. This new method has several advantages, having full automation, precise detection, and real-time labelling. This network was also trained to analyze zebrafish with cardiovascular defects (pericardial edema) induced by chemical treatments with ethanol and ponatinib. It was revealed that the heart rate, EDV/ESV, stroke volume, and cardiac output from both the ethanol and ponatinib groups displayed significant reductions compared with the control. Hopefully, this trained DLC network can contribute to a better understanding and investigation of the existing cardiovascular system and abnormalities.AbstractDeepLabCut (DLC) is a deep learning-based tool initially invented for markerless pose estimation in mammals. In this study, we explored the possibility of adopting this tool for conducting markerless cardiac physiology assessment in an important aquatic toxicology model of zebrafish (Danio rerio). Initially, high-definition videography was applied to capture heartbeat information at a frame rate of 30 frames per second (fps). Next, 20 videos from different individuals were used to perform convolutional neural network training by labeling the heart chamber (ventricle) with eight landmarks. Using Residual Network (ResNet) 152, a neural network with 152 convolutional neural network layers with 500,000 iterations, we successfully obtained a trained model that can track the heart chamber in a real-time manner. Later, we validated DLC performance with the previously published ImageJ Time Series Analysis (TSA) and Kymograph (KYM) methods. We also evaluated DLC performance by challenging experimental animals with ethanol and ponatinib to induce cardiac abnormality and heartbeat irregularity. The results showed that DLC is more accurate than the TSA method in several parameters tested. The DLC-trained model also detected the ventricle of zebrafish embryos even in the occurrence of heart abnormalities, such as pericardial edema. We believe that this tool is beneficial for research studies, especially for cardiac physiology assessment in zebrafish embryos.Keywords:DeepLabCut;zebrafish;cardiac physiology;deep learning
研究领域:
生物学
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医学
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毒理学
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产品关联
DOI
PubMed
高影响因子
Biology-Basel
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2021
·Vol. 10
·期 6
·548
·
IF 5.079
Ferry Saputra,Yu-Heng Lai,Rey Arturo T. Fernandez,Allan Patrick G. Macabeo,Hong-Thih Lai,Jong-Chin Huang,Chung-Der Hsiao
Simple SummaryThe usage of artificial sweetener has been increased from year to year as the result of pursuing healthy lifestyle. However, ironically, several studies suggest that the consumption of artificial sweeteners cause sugar-related adverse effects (e.g., obesity, type 2 diabetes and cardiovascular disease). In this study, we explore the potential cardiovascular adverse effect of several artificial sweeteners using zebrafish as animal model. We found that artificial sweetener at the highest concentration found in nature only slightly alter the cardiovascular performance of zebrafish larvae. Furthermore, no alteration of cardiac performance showed after longer incubation which support the safety of artificial sweeteners.AbstractArtificial sweeteners are widely used food ingredients in beverages and drinks to lower calorie intake which in turn helps prevent lifestyle diseases such as obesity. However, as their popularity has increased, the release of artificial sweetener to the aquatic environment has also increased at a tremendous rate. Thus, our study aims to systematically explore the potential cardiovascular physiology alterations caused by eight commercial artificial sweeteners, including acesulfame-K, alitame, aspartame, sodium cyclamate, dulcin, neotame, saccharine and sucralose, at the highest environmentally relevant concentration on cardiovascular performance using zebrafish (Danio rerio) as a model system. Embryonic zebrafish were exposed to the eight artificial sweeteners at 100 ppb and their cardiovascular performance (heart rate, ejection fraction, fractional shortening, stroke volume, cardiac output, heartbeat variability, and blood flow velocity) was measured and compared. Overall, our finding supports the safety of artificial sweetener exposure. However, several finding like a significant increase in the heart rate and heart rate variability after incubation in several artificial sweeteners are noteworthy. Biomarker testing also revealed that saccharine significantly increase the dopamine level in zebrafish larvae, which is might be the reason for the cardiac physiology changes observed after saccharine exposure.Keywords:artificial sweeteners;zebrafish;cardiac performance
研究领域:
毒理学
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环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
AQUATIC TOXICOLOGY
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2019
·
IF 3.794
Jian Han,Kechun Liu,Rongchun Wang,Yun Zhang,Bingsheng Zhou
Otolith consisting largely of calcium carbonate, fibrous and proteins, is vital for maintaining body balance and/or hearing of fish. The formation of otolith involves Ca 2+ transport and deposition. In the present study, we investigated the effects of Cd 2+ on otoliths development by using zebrafish embryos as model. The results showed that exposure to Cd 2+ inhibited the utricular and saccular otoliths growth, indicated by reduced lateral areas. Swimming speeds were reduced and a losing balance control was observed in Cd 2+ exposed larvae. The genes related to Ca 2+ transport (e.g. plasma membrane Ca 2+ -ATPase isoform 2 , pmca2 ; Ca 2+ -ATPase isoform 2 , atp2b1a ) and regulation (e.g. parathyroid hormone ligand type-1 , pth1 ; stanniocalcin isoform 1 , stc1 ) were significantly downregulated. However, the adverse effects of Cd 2+ on otoliths growth and swimming activity can be protected by supplementation of Ca 2+ in exposure medium. Body burden of Cd 2+ in larvae was reduced upon the supplement with Ca 2+ . The overall results suggest that exposure to Cd 2+ can inhibit influx of Ca 2+ , leading to less deposition of CaCO 3 for otolith growth, and finally result in impaired balance control and swimming activity in zebrafish larvae.
研究领域:
环境毒理学
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发育生物学
✓ 已验证
DOI
PubMed
高影响因子
FISH & SHELLFISH IMMUNOLOGY
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2017
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IF 3.148
Chao Wang,Yan-Ling Chen,Wan-Ping Bian,Shao-Lin Xie,Ge-Le Qi,Li Liu,Phyllis R. Strauss,Ji-Xing Zou,De-Sheng Pei
Myostatin (Mstn) is a negative regulator of muscle development in vertebrates. Although its function in muscle growth has been well studied in mammals and fish, it remains unclear whether or how mstn functions in the immune system. In this study, mstna −/- and mstnb −/- homozygous zebrafish were firstly generated using CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9). Deletion of mstnb but not mstna enhanced growth performance. Although survival rates under normal conditions were slightly decreased in both strains, mortality after dexamethasone-induced stress was increased by ∼30%. Furthermore, transcriptional levels of several critical immune-related genes were decreased, and the ability to withstand exposure to pathogenic E. tarda was decreased, compared with that of controls. In mstnb −/- but not mstna −/- zebrafish, expression of NF-κB subunits and several pro-inflammatory cytokines failed to respond to E. tarda exposure except nfkb1 , c-rel and tnfα . Taken together, these results indicate that mstnb but not mstna plays a key role in zebrafish muscle growth. While each paralogue contributes to the response to bacterial insult, mstnb affects the immune system through activation of the NF-κB pathway, and mstna is likely to act upstream of NF-κB at some as yet unidentified target.
研究领域:
分子生物学
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免疫学
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水产养殖学
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产品关联
DOI
PubMed
高影响因子
Genes
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2021
·Vol. 12
·期 8
·1287
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IF 4.096
Shouxiang Sun,Yumei Wang,Pei-Tian Goh,Mónica Lopes-Marques,L. Filipe C. Castro,Óscar Monroig,Meng-Kiat Kuah,Xiaojuan Cao,Alexander Chong Shu-Chien,Jian Gao
Elongation of very long-chain fatty acid (Elovl) proteins are key enzymes that catalyze the rate-limiting step in the fatty acid elongation pathway. The most recently discovered member of the Elovl family, Elovl8, has been proposed to be a fish-specific elongase with two gene paralogs described in teleosts. However, the biological functions of Elovl8 are still to be elucidated. In this study, we showed that in contrast to previous findings,elovl8is not unique to teleosts, but displays a rather unique and ample phylogenetic distribution. For functional determination, we generatedelovl8a(elovl8a−/−) andelovl8b(elovl8b−/−) zebrafish using CRISPR/Cas9 technology. Fatty acid composition in vivo and zebrafish liver cell experiments suggest that the substrate preference of Elovl8 overlapped with other existing Elovl enzymes. Zebrafish Elovl8a could elongate the polyunsaturated fatty acids (PUFAs) C18:2n-6 and C18:3n-3 to C20:2n-6 and C20:3n-3, respectively. Along with PUFA, zebrafish Elovl8b also showed the capacity to elongate C18:0 and C20:1. Gene expression quantification suggests that Elovl8a and Elovl8b may play a potentially important role in fatty acid biosynthesis. Overall, our results provide novel insights into the function of Elovl8a and Elovl8b, representing additional fatty acid elongases not previously described in chordates.Keywords:elovl8;gene knockout;zebrafish;fatty acid synthesis
研究领域:
分子生物学
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遗传学
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生物化学
✓ 已验证
DOI
PubMed
高影响因子
MOLECULES
·
2019
·Vol. 24
·期 16
·2906
·
IF 3.06
Da-Le Guo,Jin-Feng Chen,Lu Tan,Meng-Ying Jin,Feng Ju,Zhi-Xing Cao,Fang Deng,Li-Na Wang,Yu-Cheng Gu,Yun Deng
Two new terpene glycosides (1–2) along with two known analogs (3–4) were obtained from the root ofSanguisorba officinalis,which is a common traditional Chinese medicine (TCM). Their structures were elucidated by nuclear magnetic resonance (NMR), electrospray ionization high resolution mass spectrometry (HRESIMS), and a hydrolysis reaction, as well as comparison of these data with the literature data. Compounds1–4exhibited anti-inflammatory propertiesin vitroby attenuating the production of inflammatory mediators, such as nitric oxide (NO) as well as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). An anti-inflammatory assay based on the zebrafish experimental platform indicated that compound1had good anti-inflammatory activity in vivo by not only regulating the distribution, but also by reducing the amount of the macrophages of the zebrafish exposed to copper sulfate.Keywords:terpene glycosides;structural elucidation;Sanguisorba officinalis;zebrafish;anti-inflammatory
研究领域:
药理学
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化学
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生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CLEAN-Soil Air Water
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2015
·Vol. 44
·期 1
·72-77
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IF 1.945
Qun He,Ruijuan Qu,Xinghao Wang,Zhongbo Wei,Ping Sun,Zunyao Wang
Although it has been widely known that arsenic poses great threat to organisms and arsenic species account for the toxic effect of the metalloid, relatively little attention has been given to the arsenic toxicity to different organisms at different pH values. Three kinds of aquatic organisms including Photobacterium phosphoreum , Daphnia magna , and Danio rerio were used to study the effects of pH on the toxicity of arsenic. Toxicity test results were analyzed by calculating EC 50 values and for arsenic species at five different pH values (5.0, 6.0, 7.0, 8.0, and 9.0). As a result, As(III) is more toxic to the tested organisms than As(V), and P. phosphoreum was more sensitive to the toxicity of arsenic than D. magna and D. rerio . The toxicity of As(III) species is different for different organisms: dihydrogen arsenite, H 2 AsO 3 – , is more toxic to P. phosphoreum and D. rerio than arsenous acid, H 3 AsO 3 , while H 3 AsO 3 is more toxic to D. magna . As for As(V), arsenate, HAsO 4 2− , is the most toxic to P. phosphoreum , whereas dihydrogen arsenate, H 2 AsO 4 – , may be responsible for the toxicity to D. magna and D. rerio . As(III) and As(V) are more toxic to P. phosphoreum in an alkaline environment, while for D. magna in an acidic environment and As(III) is more toxic to D. rerio in an alkaline environment and As(V) in an acidic environment.
研究领域:
环境生物学
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毒理学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2016
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IF 3.055
Zhong-Yan Zhou,Li-Yun Huan,Wai-Rong Zhao,Nuo Tang,Yu Jin,Jing-Yi Tang
Ethnopharmacological relevance Spatholobi Caulis is a traditional blood-activating and stasis-dispelling herb medicine, which has been used to treat diseases related to blood stasis syndrome (BSS) by inhibiting platelet aggregation, stimulate hematopoiesis, etc. It has been demonstrated that pro-angiogenesis could improve BSS. However, the pro-angiogenic activity of Spatholobi Caulis was not well elucidated Aim of study To determine the potential pro-angiogenic activity of Spatholobi Caulis and elucidate its underlying mechanism. The active fractions of Spatholobi Caulis were further screened. Material and methods Gelatin precipitation and reversed-phase liquid chromatography (RPLC) were used to purify the methanol extracts of Spatholobi Caulis , respectively. The RPLC was also used to prepare fractions. Total flavonoids of purified methanol extracts of Spatholobi Caulis (PSC) were determined using ultraviolet spectrophotometry. The morphological observation of subintestinal vessel plexus (SIVs) and tyrosine kinase inhibitor II (VRI)-induced intersegmental blood vessels (ISVs) loss in transgenic zebrafish Tg(fli-1a: EGFP)y1 were selected to evaluate the pro-angiogenic activity of PSC in vivo. Cell proliferation by MTT assay and cell migration assay were used to evaluate the pro-angiogenesis effect of PSC in human umbilical vein endothelial cells (HUVECs) in vitro . Both zebrafish and HUVECs were used in screening active fractions of PSC. The mechanism of PSC promoting angiogenesis were studied by real-time PCR in zebrafish and western blotting in HUVECs. Results Co-treatment PSC dramatically rescued VRI-induced ISVs loss in zebrafish embryos in a dose-dependent manner and 80% of the defective vascular recovered at the concentration of 30 μg/ml compared with VRI-only group. PSC also concentration-dependently increased average sprouting number and diameter of SIVs in zebrafish embryo. Real-time PCR assay proved that PSC significantly restored the down regulation of VEGFRs including Flt-1, Kdr and Kdrl induced by VRI in zebrafish (P<0.001). Furthermore, PSC not only promoted proliferation and migration of normal HUVECs but also ameliorated VRI-induced HUVECs cytotoxicity. Western blotting assay showed that co-treatment of PSC increased the expression of VEGFRs and phosphorylation of MAPKs which decreased by VRI treatment. In addition, quality evaluation experiments showed that the content of total flavonoids of PSC reached 56.36% and the main pro-angiogenic fractions of PSC were F3, F4 and F5 both in zebrafish and HUVECs. Conclusions Our data demonstrated that PSC presented pro-angiogenic activity both in zebrafish and HUVECs, and principal pro-angiogenic active components were likely flavonoids. Thus, the current study provided evidence for the clinical usage of Spatholobi Caulis in promoting blood circulation and removing stasis in traditional Chinese medicine (TCM).
研究领域:
民族药理学
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分子生物学
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心血管研究
✓ 已验证
DOI
PubMed
高影响因子
LIFE SCIENCES
·
2024
·
IF 5.2
Xuehan Dou,Zhongyuan Chen,Yunzhang Liu,Yun Li,Junli Ye,Ling Lu
Aims Prolyl hydroxylase domain 2 (PHD2), encoded by the Egln1 gene, serves as a pivotal regulator of the hypoxia-inducible factor (HIF) pathway and acts as a cellular oxygen sensor. Somatic inactivation of Phd2 in mice results in polycythemia and congestive heart failure. However, due to the embryonic lethality of Phd2 deficiency, its role in development remains elusive. Here, we investigated the function of two egln1 paralogous genes, egln1a and egln1b , in zebrafish. Main methods The egln1 null zebrafish were generated using the CRISPR/Cas9 system. Quantitative real-time PCR assays and Western blot analysis were employed to detect the effect of egln1 deficiency on the hypoxia signaling pathway. The hypoxia response of egln1 mutant zebrafish were assessed by analyzing heart rate, gill agitation frequency, and blood flow velocity. Subsequently, o-dianisidine staining and in situ hybridization were used to investigate the role of egln1 in zebrafish hematopoietic function. Key findings Our data show that the loss of egln1a or egln1b individually has no visible effects on growth rate. However, the egln1a ; egln1b double mutant displayed significant growth retardation and elevated mortality at around 2.5 months old. Both egln1a - null and egln1b - null zebrafish embryo exhibited enhanced tolerance to hypoxia, systemic hypoxic response that include hif pathway activation, increased cardiac activity, and polycythemia. Significance Our research introduces zebrafish egln1 mutants as the first congenital embryonic viable systemic vertebrate animal model for PHD2, providing novel insights into hypoxic signaling and the progression of PHD2- associated disease.
研究领域:
发育生物学
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分子生物学
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遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）