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Zhonghua Yi Xue Za Zhi Jan 2016To investigate the effects of fine particulate matter with a mean aerodynamic diameter ≤2.5 μm (PM2.5) collected from Lanzhou city on phagocytic function of alveolar...
OBJECTIVE
To investigate the effects of fine particulate matter with a mean aerodynamic diameter ≤2.5 μm (PM2.5) collected from Lanzhou city on phagocytic function of alveolar macrophages (AM) in chronic obstructive pulmonary disease (COPD) mice.
METHODS
Forty male mice were randomly divided into four groups: healthy group, healthy PM2.5 group, COPD group and COPD PM2.5 group. COPD mice were established by cigarette smoking. PM2.5 (10 mg/kg) collected by air sampler was intratracheally instilled in healthy PM2.5 group and COPD PM2.5 group. Mice were sacrificed after 14 days, and alveolar macrophages (AM) were isolated. Mean fluorescence intensity (MFI) and the positive percent of alveolar macrophages engulfing flurescein isothiocyanate-labeled Escherichia coli (FITC-E.coli) (AM%) were detected by flow cytometry. Total antioxidative capacity (TAC) was measured by O-phenanthroline colorimetry. Malondialdehyde (MDA) was measured by thiobarbiturieacid colorimetry and myeloperoxidase (MPO) was measured by O-dianisidine colorimetry.
RESULTS
The peak inspiratory flow (PIF), peak expiratory flow (PEF) and dynamic compliance (Cdyn) of COPD group were significantly lower than healthy control group. The pathology of COPD group showed disruption of alveolar septa, formation of emphysema, and that the number of alveoli had a significant reduction. The MFI and AM% in COPD group were significant lower than healthy group (14.1±1.7 vs 43.2±6.1, 9.2%±2.3% vs 69.1%±8.3%)(all P<0.01). Comparing to healthy group and COPD group, the MFI and AM% in healthy PM2.5 group (20.3±4.5, 40.4%±4.4%) and COPD PM2.5 group (7.5±1.3, 6.0%±2.2%) were respectively lowered. The level of TAC in COPD group was significantly lower than healthy group [(3.10±0.64) vs (15.43±0.69)U/mg], the levels of MDA and MPO in COPD group were higher than healthy group[(2.72±0.13) vs (1.31±0.16) nmol/mg, (1.63±0.11) vs (0.92±0.13)U/g] (all P<0.01). In both healthy PM2.5 group and COPD PM2.5 group, the levels of TAC [(6.75±1.06), (2.34±0.61) U/mg] were lower than their corresponding control group; while the levels of MDA [(1.96±0.31), (3.20±0.19) nmol/mg] and the levels of MPO [(1.01±0.19), (1.74±0.13) U/g] were increased (all P<0.01). For the COPD group at baseline and after the intervention of PM2.5, the MFI and AM% showed positive correlation with the levels of TAC, and negative correlation with the levels of MDA , and negative correlation with the levels of MPO (all P<0.05). For health group at baseline and after the intervention PM2.5, the above relationships still existed (all P<0.05).
CONCLUSION
PM2.5 can damage phagocytosis of AM and exacerbate oxidative stress in COPD mice, and AM phagocytosis impairment by PM2.5 is closely associated with oxidative stress.
Topics: Animals; Lung; Macrophages, Alveolar; Male; Malondialdehyde; Mice; Oxidative Stress; Particulate Matter; Phagocytosis; Pulmonary Alveoli; Pulmonary Disease, Chronic Obstructive; Pulmonary Emphysema; Smoking; Nicotiana
PubMed: 26879794
DOI: 10.3760/cma.j.issn.0376-2491.2016.04.016 -
Chemosphere Aug 2017The risk of acetochlor to human health is still unclear, prompting concern over its risk, especially to pesticide suicides population, occupational population (farmers,...
The risk of acetochlor to human health is still unclear, prompting concern over its risk, especially to pesticide suicides population, occupational population (farmers, retailers and pharmaceutical workers), and special population (young children and infants, pregnant women, older people, and those with compromised immune systems). This study was to explore the toxic effect and the possible mechanism of toxic action of acetochlor using zebrafish larvae whose toxicity profiles have been confirmed to be strikingly similar with mammalian. The result indicated that the toxic target organ of acetochlor was cardiovascular system. Thus, cardiovascular toxicity evaluation was investigated systematically. The main phenotypes of cardiovascular toxicity induced by acetochlor were bradycardia, pericardial edema, circulation defect, and thrombosis; Malformed heart was confirmed by histopathological examination. Thrombosis which maybe triggered by bradycardia was further studied using o-dianisidine for erythrocyte staining; Substantial thrombus in the caudal vein and significantly reduced heart red blood cells (RBCs) intensity which can reflect the thrombosis degree were observed in zebrafish in a concentration-dependent manner. Additionally, the mRNA expression level of Nkx2.5 and Gata4 related to induction of cardiac program were down-regulated significantly by quantitative real-time polymerase chain reaction (qRT-PCR), which could cause defects in the cardiovascular system. For the first time, our results demonstrated that acetochlor induced cardiovascular toxicity, and down-regulation of Nkx2.5 and Gata4 might be its possible molecular basis. Our data generated here might provide novel insights into cardiovascular disease risk following acetochlor exposure to human, especially to pesticide suicides population, occupational population and special population.
Topics: Animals; Cardiovascular System; Down-Regulation; GATA Transcription Factors; Herbicides; Homeobox Protein Nkx-2.5; Humans; Larva; RNA, Messenger; Toluidines; Zebrafish; Zebrafish Proteins
PubMed: 28472748
DOI: 10.1016/j.chemosphere.2017.04.090 -
Zebrafish Oct 2019Strategies to reduce reperfusion injury after ischemia have been considered in clinical practice, but few interventions have successfully passed the proof-of-concept...
Strategies to reduce reperfusion injury after ischemia have been considered in clinical practice, but few interventions have successfully passed the proof-of-concept stage. In this study, we developed a novel zebrafish larvae hypoxia/reoxygenation (H/R) model to simulate myocardial ischemia/reperfusion injury (MIRI), with potential utility as a drug screening tool. After H/R treatment, videos of transgenic [Tg(cmlc:EGFP)] larval zebrafish hearts were captured using a digital high-speed camera, and the heart rate, diastolic area, systolic area, and total fraction of area changed were quantified. The mRNA expression of and was quantified, and red blood cells (RBCs) were detected by O-dianisidine staining. We found that a decline in cardiac contractility occurred in zebrafish larvae 48 h after hypoxia treatment. Reoxygenation for 2-5 h after 48 h of hypoxia caused heart dysfunction in zebrafish larvae, and were determined to be the optimum conditions for simulating MIRI similar to mammalian models. Our results indicated that heart dysfunction after reoxygenation in zebrafish larvae was accompanied by an upregulated gene expression of a number of myocardial injury biomarkers and increased numbers of RBCs. In conclusion, the novel larval zebrafish H/R model developed in this study could be used for rapid screening and efficacy assessment of MIRI therapeutics.
Topics: Animals; Animals, Genetically Modified; Green Fluorescent Proteins; Hypoxia; Larva; Myocardial Contraction; Myocardial Reperfusion Injury; Oxygen; Zebrafish
PubMed: 31314708
DOI: 10.1089/zeb.2018.1722 -
Aquatic Toxicology (Amsterdam,... May 2021Exposure to oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) at critical developmental time-points in fish models impairs red blood cell concentrations in a...
Exposure to oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) at critical developmental time-points in fish models impairs red blood cell concentrations in a regioselective manner, with 2-hydroxychrysene being more potent than 6-hydroxychrysene. To better characterize this phenomenon, embryos of Japanese medaka (Oryzias latipes) were exposed to 2- or 6-hydroxychrysene (0.5, 2, or 5 μM) from 4 h-post-fertilization (hpf) to 7 d-post-fertilization. Following exposure, hemoglobin concentrations were quantified by staining fixed embryos with o-dianisidine (a hemoglobin-specific dye) and stained embryos were imaged using brightfield microscopy. Exposure to 2-hydroxychrysene resulted in a concentration-dependent decrease in hemoglobin relative to vehicle-exposed embryos, while only the highest concentration of 6-hydroxychrysene resulted in a significant decrease in hemoglobin. All tested concentrations of 2-hydroxychrysene also caused significant mortality (12.2 % ± 2.94, 38.9 % ± 14.4, 85.6 % ± 11.3), whereas mortality was not observed following exposure to 6-hydroxychrysene. Therefore, treatment of embryos with 2-hydroxychrysene at various developmental stages and durations was subsequently conducted to identify key developmental landmarks that may be targeted by 2-hydroxychrysene. A sensitive window of developmental toxicity to 2-hydroxychrysene was found between 52-100 hpf, with a 24 h exposure to 10 μM 2-hydroxychrysene resulting in significant anemia and mortality. Since exposure to 2-hydroxychrysene from 52 to 100 hpf, a window that includes liver morphogenesis in medaka, resulted in the highest magnitude of toxicity, liver development and function may have a role in 2-hydroxychrysene developmental toxicity.
Topics: Animals; Chrysenes; Embryo, Nonmammalian; Embryonic Development; Hemoglobins; Oryzias; Stereoisomerism; Water Pollutants, Chemical
PubMed: 33714762
DOI: 10.1016/j.aquatox.2021.105791 -
Journal of the Science of Food and... Aug 2017Peroxidase activity was increased during germination of green gram and such an increase may have benefits in many physiological processes. The present study aimed to...
BACKGROUND
Peroxidase activity was increased during germination of green gram and such an increase may have benefits in many physiological processes. The present study aimed to investigate the optimum conditions for the extraction, purification and characterization of peroxidase from the germinated green gram roots and also its application for the removal of phenols in water.
RESULTS
Peroxidase activity was increased by 300-fold in 5-day germinated green gram. Because the root was rich in peroxidase activity, peroxidase from roots was isolated and purified to homogeneity. The purified peroxidase showed a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis with a molecular weight of 50 kDa, an optimum pH of 5.5 and a pH stability ranging from 5 to 9. The enzyme had 50% residual activity at 70 °C. It catalyzed the oxidation of a variety of substrates. The K value of the enzyme was 1.28 mmol L for o-dianisidine and 0.045 mmol L for H O . The enzyme lost 100% activity in the presence of dithiothreitol and cysteine. The addition of copper ion increased the enzyme activity by three-fold. Both soluble and immobilized peroxidases removed more phenol than p-chlorphenol, whereas horseradish peroxidase removed more p-chlorphenol. Thus, the green gram root peroxidase showed good pH and temperature stability, as well as the ability to remove phenolic compounds from effluent.
CONCLUSION
Peroxidase with good thermal and pH stability was purified from germinated green gram roots and has the ability to oxidize phenolic compounds from waste water. © 2016 Society of Chemical Industry.
Topics: Chlorophenols; Enzyme Stability; Enzymes, Immobilized; Hydrogen-Ion Concentration; Kinetics; Peroxidase; Phenol; Plant Proteins; Plant Roots; Vigna; Water Pollutants, Chemical
PubMed: 27976372
DOI: 10.1002/jsfa.8173 -
Clinical Chemistry and Laboratory... Aug 2017Meta-analyses indicated the breakdown of copper homeostasis in the sporadic form of Alzheimer's disease (AD), comprising copper decreases within the brain and copper...
BACKGROUND
Meta-analyses indicated the breakdown of copper homeostasis in the sporadic form of Alzheimer's disease (AD), comprising copper decreases within the brain and copper increases in the blood and the pool not bound to ceruloplasmin (non-Cp Cu, also known in the literature as "free" copper). The calculated non-Cp Cu (Walshe's) index has many limitations.
METHODS
A direct fluorescent method for non-Cp Cu detection has been developed and data are presented herein. The study included samples from 147 healthy subjects, 36 stable mild cognitive impairment (MCI) and 89 AD patients, who were tested for non-Cp Cu through the direct method, total serum copper, ceruloplasmin concentration and o-dianisidine ceruloplasmin activity. The indirect non-Cp Cu Walshe's index was also calculated.
RESULTS
The direct method was linear (0.9-5.9 μM), precise (within-laboratory coefficient variation of 9.7% for low and 7.1% for high measurements), and had a good recovery. A reference interval (0-1.9 μM) was determined parametrically in 147 healthy controls (27-84 years old). The variation of non-Cp Cu was evaluated according to age and sex. Non-Cp Cu was 1.5 times higher in AD patients (regarding the upper value of the reference interval) than in healthy controls. Healthy, MCI and AD subjects were differentiated through the direct non-Cp Cu method [areas under the curve (AUC)=0.755]. Considering a 95% specificity and a 1.91 μmol/L cut-off, the sensitivity was 48.3% (confidence interval 95%: 38%-58%). The likelihood ratio (LR) was 9.94 for positive test results (LR+) and 0.54 for negative test result (LR-).
CONCLUSIONS
The direct fluorescent test reliably and accurately measures non-Cp Cu, thereby determining the probability of having AD.
Topics: Adult; Aged; Aged, 80 and over; Alzheimer Disease; Copper; Female; Fluorescent Dyes; Humans; Male; Middle Aged; Spectrometry, Fluorescence
PubMed: 28076308
DOI: 10.1515/cclm-2016-0843 -
Aquatic Toxicology (Amsterdam,... Jun 2016Copper, as an essential trace mineral, can cause diseases such as childhood leukemia at excess levels, but has been applied in anemia therapy for a long time. However,...
Copper, as an essential trace mineral, can cause diseases such as childhood leukemia at excess levels, but has been applied in anemia therapy for a long time. However, few reports have studied its role during hematopoiesis at the molecular level in an animal model. In this study, by microarray, qRT-PCR, whole-mount in situ hybridization and O-dianisidine staining detections, we revealed the increased expression of hemoglobin in copper-exposed embryos. Secondly, we found that copper-exposed embryos exhibited high levels of reactive oxygen species (ROS), and genes in oxygen binding and oxygen transporting were up-regulated in the embryos. Finally, we found that ROS scavengers NAC, GSH, and DMTU not only inhibited in vivo ROS levels induced by copper, but also significantly decreased high expression of hemoglobin back to almost normal levels in copper exposed embryos, and also helped with copper elimination from the embryos. Our data first demonstrated that ROS mediated copper induced hemoglobin expression in vertebrates, partly revealing the underlying molecular mechanism of copper therapy for anemia. Moreover, we revealed that copper homeostasis was broken by its induced ROS and ROS helped with copper overloading in the body, which could be applied as a novel therapy target for copper-caused diseases.
Topics: Analysis of Variance; Animals; Copper; Gene Expression Regulation; Hemoglobins; Mass Spectrometry; Microarray Analysis; Polymerase Chain Reaction; Reactive Oxygen Species; Reverse Transcriptase Polymerase Chain Reaction; Up-Regulation; Water Pollutants, Chemical; Zebrafish
PubMed: 26991749
DOI: 10.1016/j.aquatox.2016.03.008 -
Comparative Biochemistry and... Jul 2023Sesamin, the major lignan in sesame seeds (Sesamum indicum L.), is known to have several pharmaceutical activities. However, its toxicological profile is still limited,...
Sesamin, the major lignan in sesame seeds (Sesamum indicum L.), is known to have several pharmaceutical activities. However, its toxicological profile is still limited, especially regarding embryotoxicity. This study aimed to evaluate the developmental toxicity of sesamin in zebrafish embryos. After 72 h exposure, sesamin did not affect the survival and hatching rates, nor did it cause malformation in zebrafish embryos. Cardiotoxicity was also evaluated by monitoring embryo heartbeats and erythrocyte staining using o-dianisidine. The results showed that sesamin did not affect heart morphology, heart rate, or cardiac output in zebrafish embryos. The present study also evaluated sesamin's anti-angiogenesis, antioxidant and anti-inflammation activities. Sesamin significantly decreased the sub-intestinal vessel plexus as revealed by alkaline phosphatase staining indicating the compound exhibited anti-angiogenesis activity. For the antioxidant and anti-inflammatory assays, oxidative stress and inflammation in zebrafish embryos were induced by hydrogen peroxide and lipopolysaccharide, respectively. The reactive oxygen species (ROS) and nitric oxide (NO) production were detected using a fluorescent dye. Sesamin significantly decreased ROS and NO production in zebrafish embryos. In addition, the transcription examination by qRT-PCR of oxidative- and inflammation-related genes showed that sesamin affected the genes in a manner that correlated with results from the efficacy assays. In conclusion, the present study revealed that sesamin did not cause embryotoxicity and cardiotoxicity in zebrafish embryos. In addition, it exhibited evidence of anti-angiogenesis, antioxidant and anti-inflammatory activities.
Topics: Animals; Zebrafish; Antioxidants; Reactive Oxygen Species; Cardiotoxicity; Oxidative Stress; Lignans; Inflammation; Anti-Inflammatory Agents; Embryo, Nonmammalian
PubMed: 37098389
DOI: 10.1016/j.cbpc.2023.109637 -
Journal of Applied Toxicology : JAT Feb 2024Angiogenesis and hemodynamic instability created by the irregular blood vessels causes hypoperfusion and angiogenesis-mediated diseases. Therefore, therapies focusing on...
Angiogenesis and hemodynamic instability created by the irregular blood vessels causes hypoperfusion and angiogenesis-mediated diseases. Therefore, therapies focusing on controlling angiogenesis will be a valuable approach to treat a broad spectrum of diseases. In this study, we explored the anti-angiogenic potential of berberine (BBR) and also analyzed blood flow hemodynamics using zebrafish embryos. Zebrafish embryos treated with BBR (0.01-0.75 mM) at various doses at 1 hour post-fertilization (hpf) developed a variety of phenotypic variations including aberrant blood vessels, tail bending, edema, and hemorrhage. Survival rates were much lower at higher dosages, and hatching rates were almost 99%, whereas control group appeared normal. Heart rate is an essential measure that has a strong association with hemodynamics. We used ImageJ software to study the heart rate of embryos treated with BBR, preceded by video processing. The resultant graph shows a significant decrease in heart rate of embryos treated with BBR in dose-dependent manner. Also, RBC staining using o-Dianisidine confirms the anti-angiogenic potential of BBR by indicating the decrease in the intersegmental vessels at 0.5 and 0.75 mM treated embryos. Further, the gene expression study determined that the transcripts (vegf, vegfr2, nrp1a, hif-1α, nos2a, nos2b, cox-2a, and cox-2b) measured were found to be downregulated by BBR at 0.5 mM concentration, from which we conclude that enos/vegf signaling could play an important role in modulating angiogenesis. Our data imply that BBR may be an effective compound for suppressing angiogenesis in vivo, which might be helpful in the treatment of vascular disorders like cancer and diabetic retinopathy in future.
Topics: Animals; Zebrafish; Berberine; Vascular Endothelial Growth Factor A; Angiogenesis; Hemodynamics
PubMed: 37615217
DOI: 10.1002/jat.4529 -
Frontiers in Cell and Developmental... 2022Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development...
Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development and circulatory failure, suggesting additional roles for TF in embryonic development beyond coagulation. In this study, we characterized the role of one of the TF paralogs () using a zebrafish model. The expression of during embryonic developmental stages was determined by RT-PCR. Spatiotemporal expression pattern of revealed (high expression from 28 to 36 hpf) the role of in the development of the yolk sac, circulation, and fins. Morpholinos (MO), an antisense-based oligonucleotide strategy, was used to knockdown and examined for defects in morphological appearance, bleeding, and vascular patterning. MO-injected embryos showed morphological abnormalities, including shorter body lengths and crooked tails. O-dianisidine staining showed MO-injected embryos exhibited bleeding in the trunk (5.44%) and head (9.52%) regions. Imaging of endothelial-specific transgenic lines () showed a 3-fold decreased caudal vein plexus (CVP) in morphants versus controls at 48 hpf, suggesting a potential role for in angiogenesis. These findings confirm that is essential for angiogenesis, in addition to its involvement in hemostasis.
PubMed: 35386206
DOI: 10.3389/fcell.2022.852989