OBJECTIVES: To investigate the role and mechanism of copper overload-mediated endoplasmic reticulum stress (ERS) in vascular endothelial injury in Kawasaki disease (KD). METHODS: Four-week-old male C57BL/6 mice were randomly divided into four groups: control, KD, KD plus copper chelator tetrathiomolybdate (TTM), and KD plus ERS inhibitor AMG PERK 44 (AMG) (n=20 per group). A KD mouse model was established using Candida albicans extract. Human umbilical vein endothelial cells (HUVECs) were divided into control (intervention with healthy children's serum), KD (intervention with KD patients' serum), and KD+TTM (intervention with KD patients' serum plus 20 µmol/L TTM). Copper deposition in mouse heart tissue was assessed using rubeanic acid staining. Vascular pathological changes were observed using hematoxylin-eosin staining and measurement of abdominal aortic diameter and area. ERS activation was detected by transmission electron microscopy and immunofluorescence. HUVEC viability, apoptosis, and functional changes were evaluated using CCK8, flow cytometry, cell scratch assay, and angiogenesis experiments. ERS marker protein expression levels were measured by Western blot. RESULTS: Compared to the KD group, the KD+TTM and KD+AMG groups showed reduced copper deposition in the vascular wall, decreased swelling of coronary endothelial cells and endoplasmic reticulum, reduced inflammatory cell infiltration, and less abdominal aortic lesion expansion. The abdominal aortic diameter and area, and the fluorescence intensity of ERS marker proteins (GRP78 and CHOP) were significantly lower (P<0.05). Compared to the KD group, the KD+TTM group exhibited increased cell viability, tube number, and scratch healing rate, along with decreased apoptosis rate and expression of ERS marker proteins (GRP78, CHOP, ATF6, and p-PERK) (P<0.05). CONCLUSIONS Copper overload aggravates vascular endothelial injury in KD by activating the ERS pathway. TTM can exert protective effects on the endothelium by regulating copper metabolism and inhibiting the ERS pathway.
Endoplasmic Reticulum Stress ; Copper/toxicity* ; Male ; Mucocutaneous Lymph Node Syndrome/metabolism* ; Animals ; Humans ; Endoplasmic Reticulum Chaperone BiP ; Mice, Inbred C57BL ; Mice ; Human Umbilical Vein Endothelial Cells ; Apoptosis ; Endothelium, Vascular/injuries*

OBJECTIVES: To explore the mechanisms of Qingre Lidan Jiedu Recipe (QLJR) for improving cognitive dysfunction in rats with high copper load. METHODS: Seventy-five male SD rats were randomized into normal control group, model group, QLJR group, penicillamine (PCA) group, and QLJR+ PCA group. Except for those in the control group, all the rats were fed a high-copper diet for 12 weeks. The effects of the treatments on cognitive function of the rats were assessed using the Barnes maze and passive avoidance tests. Hippocampal expressions of NIX, FUNDC1 and LC3 of the rats were detected using Western blotting and immunofluorescence staining, and changes in mitochondrial morphology were observed with transmission electron microscopy. RESULTS: Behavioral tests showed prolonged target hole latency, shortened latency to enter the dark chamber, and increased error counts of the rats in the model group, which were significantly improved in QLJR+PCA group; the error counts were significantly lower in QLJR+PCA group than in either QLJR or PCA group. Among all the groups, the hippocampal expressions of NIX and FUNDC1 were the lowest and LC3 I/II expression the highest in the model group; NIX and FUNDC1 expressions were significantly higher and LC3 I expression was lower in QLJR+PCA group than in QLJR group and PCA group. Immunofluorescence staining revealed weakened NIX and FUNDC1 expressions and enhanced LC3 expression in the hippocampus of the rats in the model group as compared with those in the normal control and QLJR+PCA groups, but their expressions did not differ significantly between QLJR and PCA groups. The rats in the model group showed obvious structural disarray of the mitochondria, which were improved in all the treatment groups. CONCLUSIONS QLJR improves cognitive dysfunction in rats with high copper load possibly by regulating mitophagy.
Animals ; Male ; Rats, Sprague-Dawley ; Rats ; Drugs, Chinese Herbal/therapeutic use* ; Copper/toxicity* ; Mitophagy/drug effects* ; Hippocampus/drug effects* ; Cognition Disorders/drug therapy* ; Cognitive Dysfunction/chemically induced*

Animals ; Copper ; toxicity ; Disulfiram ; toxicity ; Humans ; Leukemia, Myeloid, Acute ; Mice ; Mice, Inbred NOD ; Mice, SCID

OBJECTIVETo investigate the effects of lead exposure on the copper concentration in the brain and serum and the expression of copper transporters in the choroid plexus among rats.

METHODSSixty specific pathogen-free Sprague-Dawley rats were randomly divided into a control group and three lead-exposed groups, with 8 mice in each group. The lead-exposed groups were orally administrated with 500 (low-dose group)), 1 000 (middle-dose group), and 2 000 mg/L (high-dose group) lead acetate in drinking water for eight weeks. And the rats in control group were given 2 000 mg/L sodium acetate in drinking water. The content of lead and copper in the serum, hippocampus, cortex, choroid plexus, bones, and cerebrospinal fluid (CSF) was determined by inductively coupled plasma-mass spectrometry (ICP-MS). Confocal and real-time PCR methods were applied to measure the expression of copper transporters including copper transporter 1 (Ctr1), antioxidant protein 1 (ATX1), and Cu ATPase (ATP7A).

RESULTSCompared with the control group, the lead-exposed groups showed significantly higher lead concentrations in the serum, cortex, hippocampus, choroid plexus, CSF, and bones (P < 0.05) and significantly higher copper concentrations in the CSF, choroid plexus, serum, and hippocampus (P < 0.05). Confocal images showed that Ctr1 protein was expressed in the cytoplasm and cell membrane of choroid plexus in control group. However, Ctr1 migrated to CSF surface microvilli after lead exposure. Ctr1 fluorescence intensity gradually increased with increasing dose of lead, except that the middle-dose group had a higher Ctr1 fluorescence intensity than the high-dose group. In addition, the middle- and high-dose groups showed a lower ATX1 fluorescence intensity compared with the control group. Real-time PCR data indicated that the three lead-exposed groups showed significantly higher mRNA levels of Ctr1 and ATP7A compared with the control group (P < 0.05).

CONCLUSIONCopper homeostasis in the choroid plexus is affected by lead exposure to induce copper homeostasis disorders in brain tissue, which may be one of the mechanisms of lead neurotoxicity.

Adenosine Triphosphatases ; Animals ; Brain ; Cation Transport Proteins ; drug effects ; Choroid Plexus ; drug effects ; metabolism ; Copper ; metabolism ; Homeostasis ; Organometallic Compounds ; toxicity ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

Animals ; Bone and Bones ; drug effects ; Copper ; pharmacology ; therapeutic use ; Fluoride Poisoning ; prevention & control ; Fluorides ; blood ; toxicity ; urine ; Lipid Metabolism ; drug effects ; Molybdenum ; toxicity ; Rabbits ; Trace Elements ; pharmacology ; therapeutic use

OBJECTIVETo investigate the effect of lead exposure on copper and copper metalloenzyme and the intervention effect of quercetin.

METHODSTwenty-four specific pathogen-free male Sprague-Dawley rats of good health were randomly divided into control group (n = 8), lead acetate group (n = 8), and lead acetate + quercetin group (n = 8). The rats in lead acetate group were poisoned by drinking water with 1 g/L lead acetate for 8 weeks, while the rats in control group were fed by drinking water with sodium acetate of the same volume for 8 weeks; the rats in lead acetate+quercetin group were intraperitoneally injected with quercetin (30 mg × kg-1 × d-1) for 8 weeks while drinking water with lead acetate. The Morris water maze was used to test the learning and memory abilities of rats. The lead and copper levels in the serum, hippocampus, cortex, and bone were measured by graphite furnace atomic absorption spectrometry. The level of advanced glycation end products, activity of Cu/Zn superoxide dismutase (SOD), and content and activity of ceruloplasmin (CP) in the hippocampus and serum were measured using a test kit. HE staining was performed to observe the pathological changes in the hippocampus.

RESULTSThe Morris water maze test showed that the latency in lead acetate group (52.50±12.04 s) was significantly longer than that in control group (28.08±7.31 s) (P<0.05), and the number of platform crossings was significantly lower in the lead acetate group than in the control group. Compared with those in the control group, the lead levels in the cortex and hippocampus in lead acetate group increased 2.72-fold and 3.79-fold, and the copper in the cortex and hippocampus, and serum free copper levels in lead acetate group increased 1.15-fold, 1.48-fold, and 6.44-fold. Compared with the control group, the lead acetate group had a lower content of CP in the hippocampus (1.23±0.40 U/mg provs0.78±0.08 U/mg pro) and 31.81%and 19.49%decreases in CP content and Cu/Zn SOD activity. Free copper level in serum was positively correlated with the latency and lead levels in the serum, cortex, and hippocampus. The escape latency of rats in lead acetate + quercetin group was decreased by 42.15% (P<0.05). The lead levels in the cortex and hippocampus in lead acetate + quercetin group (0.246 ± 0.58 µg/g and 0.202±0.049 µg/g) were significantly lower than those in lead acetate group (0.391±0.49 µg/g and 0.546±0.120 µg/g), but the free copper and copper levels in the hippocampus and cortex were not significantly reduced. The lead acetate + quercetin group had higher Cu/Zn SOD activity and CP content in the hippocampus than the lead acetate group (P < 0.05). The light microscope observation showed that the number of cells in the hippocampus was reduced with disordered arrangement in the lead acetate group; with quercetin intervention, the hippocampus damage was reduced.

CONCLUSIONLead exposure results in disorder of copper homeostasis, while quercetin may alleviate the damage induced by lead to some extent.

Animals ; Cerebral Cortex ; chemistry ; Copper ; blood ; Hippocampus ; chemistry ; Homeostasis ; Learning ; drug effects ; Male ; Memory ; drug effects ; Organometallic Compounds ; toxicity ; Quercetin ; pharmacology ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the genotoxicity and oxidative stress induced by copper oxide nanoparticles in mice.

METHODSThirty mice were randomly divided into control group and low- and high-dose exposure groups. The low- and high-dose exposure groups were given copper oxide nanoparticles (50 and 150 mg/kg) by a single intraperitoneal injection, while the control group was given an equal volume of normal saline containing 0.05%Tween 80. The micronucleus rate of reticulocytes in peripheral blood from the caudal vein and urinary 8-hydroxy-deoxyguanosine (8-OH-dG) level were measured before and at 24, 48, and 72 h after exposure. All the mice were sacrificed at 72 h after exposure, the liver, kidney, and femoral marrow were taken for DNA extraction, and 8-OH-dG in DNA was quantified.

RESULTSThe micronucleus rates of peripheral blood reticulocytes in low-dose exposure group at 48 h (3.11±1.46‰ and in high-dose exposure group at 24 and 48 h (4.25±0.43) and 5.42±0.76‰) were significantly increased compared with those before exposure (1.55±0.39‰ and 1.11±0.19‰) and those in control group (1.55±0.28‰ and 1.00±0.67‰) (P < 0.05 or P < 0.01). The urinary 8-OH-dG levels (ng/mg creatinine) in low- and high-dose exposure groups at all time points were significantly increased compared with those before exposure and those in control group (P < 0.05 or P < 0.01). The low- and high-dose exposure groups had significantly higher content of 8-OH-dG in liver DNA than the control group (4.53±1.27 and 7.69±2.78 vs 0.85±0.14, P < 0.01).

CONCLUSIONCopper oxide nanoparticles cause genotoxicity and increase oxidative stress in mice.

Animals ; Copper ; toxicity ; DNA Damage ; drug effects ; Female ; Male ; Mice ; Mice, Inbred ICR ; Nanoparticles ; toxicity ; Oxidative Stress ; drug effects

OBJECTIVETo study the biological effects of nanoscale copper oxide (nCuO), zinc oxide (nZnO), cerium dioxide (nCeO2) and their mixtures on Carassius auratus.

METHODSJuvenile fish (Carassius auratus) were exposed to aqueous suspensions of nCuO, nZnO, and nCeO2 (alone and in mixtures) at concentrations of 20, 40, 80, 160, and 320 mg/L. The biomarkers-acetylcholinesterase (AChE) in brain, sodium/potassium-activated ATPase (Na+/K+-ATPase) in gill, and superoxide dismutase (SOD) and catalase (CAT) in liver-were determined after 4 days of exposure. Integrated biomarker response (IBR) was calculated by combining multiple biomarkers into a single value.

RESULTSAChE and SOD activities were significantly inhibited by all test metal oxide nanoparticles (NPs) at high concentrations (⋝160 mg/L) with the exception of nCeO2. Na+/K+-ATPase induction exhibited bell-shaped concentration-response curves. CAT activity was significantly inhibited at concentrations equal to or higher than 160 mg/L. The order of IBR values was nCeO2 ≈ nZnO/nCeO2 ≈ nCuO/nCeO2 < nCuO/nZnO/nCeO2 < nZnO < nCuO < nCuO/nZnO. The joint effect seemed to be synergistic for nCuO/nZnO mixtures, additive for the ternary mixture and less than additive or antagonistic for the binary mixtures containing nCeO2.

CONCLUSIONConcentration-dependent changes of enzymatic activities (AChE, Na+/K+-ATPase, SOD, and CAT) were observed in fish exposed to nanoscale metal oxides. IBR analysis allowed good discrimination between the different exposures and might be a useful tool for the quantification of integrated negative effects induced by NPs toward fish.

Acetylcholinesterase ; metabolism ; Animals ; Biomarkers ; metabolism ; Brain ; drug effects ; enzymology ; Cerium ; toxicity ; Copper ; toxicity ; Gills ; drug effects ; enzymology ; Goldfish ; metabolism ; Liver ; drug effects ; enzymology ; Metal Nanoparticles ; toxicity ; Random Allocation ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism ; Toxicity Tests ; Water Pollutants, Chemical ; toxicity ; Zinc Oxide ; toxicity

To study the effect of Cu2+ and Zn2+ on amyloid-beta peptides (Abeta) aggregation, the morphology, size and cell toxicity of Abeta40 aggregates formed with the metal ions have been observed by the methods including ultraviolet spectroscopy, fluorescence spectroscopy and transmission electron microscopy. The results showed that Cu2+ and Zn2+ can accelerate Abeta40 aggregation, and both changed the morphology and size of Abeta40 aggregates. Zn2+ induced Abeta40 to form fibrous Abeta40 aggregates, while the amorphous and fibrous aggregates were produced by the interaction between Cu2+ and Abeta40. In addition, H2O2 was produced when Abeta40 reduced Cu2+. The relationship between metal ions and Abeta40 aggregates was analyzed, and the function of metal ions in Alzheimer's disease (AD) was illustrated in the research.
Amyloid beta-Peptides ; chemistry ; Cell Survival ; drug effects ; Copper ; administration & dosage ; chemistry ; toxicity ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Hydrogen Peroxide ; chemistry ; Ions ; chemistry ; Microscopy, Electron, Transmission ; Peptide Fragments ; chemistry ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet ; Zinc ; chemistry

To investigate whether a series of water-soluble cross-linked chitosan derivates synthesized in the guide of imprinting technology could be used as a uranium chelating agent to protect cells exposed to depleted uranium (DU), the imprinted chitosan derivates with high UO2(2+) chelating ability were screened, and cell model of human renal proximal tubule epithelium cells (HK-2) exposed to DU (500 micromol.L-1) was built, chitosan derivates (400 mg.L-1 ) was added to test group and diethylenetriaminepentaacetic acid (DTPA, 50 mg.L-1) was added to positive control group. The results showed that three Cu2+ imprinted chitosan derivates had higher uranium chelating ability (>49 microg.mg-1) than chitosan and non-imprinted chitosan derivates. Compared to the cells exposed to DU only, survival of cells in group added chitosan derivates rose up significantly (increased from 57.3% to 88.7%, and DTPA to 72.6%), and DU intracellular accumulation decreased, membrane damage and DNA damage also eased. Among the imprinted chitosan derivates, Cu2+ imprinted penta dialdehyde cross-linked carboxymethyl chitosan (Cu-P-CMC) was the best, and better than DTPA. From ultrastructure observation, the DU precipitates of test group added Cu-P-CMC were most grouped in a big hairy clusters in a string together outside cells. It is possible that the DU-chitosan derivates precipitates are too big to enter into cells, and from this way, the DU uptake by cells decreased so as to detoxication.
Antioxidants ; metabolism ; Apoptosis ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Chelating Agents ; administration & dosage ; chemistry ; pharmacology ; Chitosan ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacology ; Copper ; chemistry ; pharmacology ; Cross-Linking Reagents ; administration & dosage ; chemistry ; pharmacology ; DNA Damage ; drug effects ; Epithelial Cells ; cytology ; ultrastructure ; Humans ; Inactivation, Metabolic ; Kidney Tubules, Proximal ; cytology ; ultrastructure ; Microscopy, Electron, Transmission ; Uranium ; toxicity ; Water