PM2.5 exposure contributes to anxiety and depression-like behaviors via phenyl-containing compounds interfering with dopamine receptor.

As a global problem, fine particulate matter (PM2.5) really needs local fixes. Considering the increasing epidemiological relevance to anxiety and depression but inconsistent toxicological results, the most important question is to clarify whether and how PM2.5 causally contributes to these mental disorders and which components are the most dangerous for crucial mitigation in a particular place. In the present study, we chronically subjected male mice to a real-world PM2.5 exposure system throughout the winter heating period in a coal combustion area and revealed that PM2.5 caused anxiety and depression-like behaviors in adults such as restricted activity, diminished exploratory interest, enhanced repetitive stereotypy, and elevated acquired immobility, through behavioral tests including open field, elevated plus maze, marble-burying, and forced swimming tests. Importantly, we found that dopamine signaling was perturbed using mRNA transcriptional profile and bioinformatics analysis, with Drd1 as a potential target. Subsequently, we developed the Drd1 expression-directed multifraction isolating and nontarget identifying framework and identified a total of 209 compounds in PM2.5 organic extracts capable of reducing Drd1 expression. Furthermore, by applying hierarchical characteristic fragment analysis and molecular docking and dynamics simulation, we clarified that phenyl-containing compounds competitively bound to DRD1 and interfered with dopamine signaling, thereby contributing to mental disorders. Taken together, this work provides experimental evidence for researchers and clinicians to identify hazardous factors in PM2.5 and prevent adverse health outcomes and for local governments and municipalities to control source emissions for diminishing specific disease burdens.

Developmental Toxicity of Fine Particulate Matter: Multifaceted Exploration from Epidemiological and Laboratory Perspectives.

Particulate matter of size ≤ 2.5 µm (PM2.5) is a critical environmental threat that considerably contributes to the global disease burden. However, accompanied by the rapid research progress in this field, the existing research on developmental toxicity is still constrained by limited data sources, varying quality, and insufficient in-depth mechanistic analysis. This review includes the currently available epidemiological and laboratory evidence and comprehensively characterizes the adverse effects of PM2.5 on developing individuals in different regions and various pollution sources. In addition, this review explores the effect of PM2.5 exposure to individuals of different ethnicities, genders, and socioeconomic levels on adverse birth outcomes and cardiopulmonary and neurological development. Furthermore, the molecular mechanisms involved in the adverse health effects of PM2.5 primarily encompass transcriptional and translational regulation, oxidative stress, inflammatory response, and epigenetic modulation. The primary findings and novel perspectives regarding the association between public health and PM2.5 were examined, highlighting the need for future studies to explore its sources, composition, and sex-specific effects. Additionally, further research is required to delve deeper into the more intricate underlying mechanisms to effectively prevent or mitigate the harmful effects of air pollution on human health.

Triazole fungicides exert neural differentiation alteration through H3K27me3 modifications: In vitro and in silico study.

Considering that humans are unavoidably exposed to triazole fungicides through the esophagus, respiratory tract, and skin contact, revealing the developmental toxicity of triazole fungicides is vital for health risk assessment. This study aimed to screen and discriminate neural developmental disorder chemicals in commonly used triazole fungicides, and explore the underlying harmful impacts on neurogenesis associated with histone modification abnormality in mouse embryonic stem cells (mESCs). The triploblastic and neural differentiation models were constructed based on mESCs to expose six typical triazole fungicides (myclobutanil, tebuconazole, hexaconazole, propiconazole, difenoconazole, and flusilazole). The result demonstrated that although no cytotoxicity was observed, different triazole fungicides exhibited varying degrees of alterations in neural differentiation, including increased ectodermal differentiation, promoted neurogenesis, increased intracellular calcium ion levels, and disturbance of neurotransmitters. Molecular docking, cluster analysis, and multiple linear regressions demonstrated that the binding affinities between triazole fungicides and the Kdm6b-ligand binding domain were the dominant determinants of the neurodevelopmental response. This partially resulted in the reduced enrichment of H3K27me3 at the promoter region of the serotonin receptor 2 C gene, finally leading to disturbed neural differentiation. The data suggested potential adverse outcomes of triazole fungicides on embryonic neurogenesis even under sublethal doses through interfering histone modification, providing substantial evidence on the safety control of fungicides.

Tebuconazole mediates cognitive impairment via the microbe-gut-brain axis (MGBA) in mice.

Tebuconazole, one of the most widely used triazole fungicides, is reported to potentially pose a risk of inducing neurological disorders in human beings. Considering the increasing exposure, whether it could influence cognitive function remains to be elucidated. Herein, we used a mouse model to evaluate the potential cognitive risks and possible mechanisms from the continuous edible application of tebuconazole at low concentrations. Our study revealed that tebuconazole deteriorated spatial learning and memory and downregulated the expression of glutamate receptor subunits. Importantly, metagenomic analysis indicated that tebuconazole not only led to significant shifts in the composition and diversity of the gut microbiota but also changed intestinal homeostasis. Specifically, after exposure, tebuconazole circulated in the bloodstream and largely entered the gut-brain axis for disruption, including disturbing the Firmicutes/Bacteroidetes ratio, interrelated neurotransmitters and systemic immune factors. Moreover, pretreatment with probiotics improved immune factor expression and restored the deterioration of synaptic function and spatial learning and memory. The current study provides novel insights concerning perturbations of the gut microbiome and its functions as a potential new mechanism by which tebuconazole exposes cognitive function-related human health.

Abnormal neural differentiation in response to graphene quantum dots through histone modification interference.

Graphene quantum dots (GQDs) have been broadly applied in biomedicine in recent years, and their environmental exposure and toxicological impacts have raised increasing concerns. The nanosafety assessment on the nervous system is one of the most important aspects, and potential effects of GQDs on neurodevelopment and the underlying mechanism are still elusive. In this study, the neural developmental toxicities of OH-GQDs and NH2-GQDs were investigated using the mouse embryonic stem cells (mESCs). The results revealed that OH-GQDs significantly inhibited the ectoderm development, and reduced the neural precursor formation and neurogenesis during the neural differentiation of the mESCs. The exploration on the mechanism uncovered that the increased enrichment of H3K27me3 at the promoter region of the Smad6 gene was involved in histone modification-activated BMP signal pathway, which consequently influenced its regulatory effects on neural differentiation. Additionally, OH-GQDs elicited a stronger effect on inducing the imbalance of histone modification, and resulted in higher latency of neural differentiation disturbance than did NH2-GQDs, suggesting surface functionalization-specific effects of GQDs on neurodevelopmental toxicity. This study would provide new insights in not only the adverse effects of GQDs on neurodevelopment, but also the influence from the chemical modification of GQDs on their bioactivities.

Perfluorinated Iodine Alkanes Promoted Neural Differentiation of mESCs by Targeting miRNA-34a-5p in Notch-Hes Signaling.

The neurodevelopmental process is highly vulnerable to environmental stress from exposure to endocrine-disrupting chemicals. Perfluorinated iodine alkanes (PFIs) possess estrogenic activities, while their potential neurodevelopmental toxicity remains blurry. In the present study, the effects of two PFIs, including dodecafluoro-1,6-diiodohexane (PFHxDI) and tridecafluorohexyl iodide (PFHxI), were investigated in the neural differentiation of the mouse embryonic stem cells (mESCs). Without influencing the cytobiological process of the mESCs, PFIs interfered the triploblastic development by increasing ectodermal differentiation, thus promoting subsequent neurogenesis. The temporal regulation of PFIs in Notch-Hes signaling through the targeting of mmu-miRNA-34a-5p provided a substantial explanation for the underlying mechanism of PFI-promoted mESC commitment to the neural lineage. The findings herein provided new knowledge on the potential neurodevelopmental toxicities of PFIs, which would help advance the health risk assessment of these kinds of emerging chemicals.

Constructing an MCF-7 breast cancer cell-based transient transfection assay for screening RARα (Ant)agonistic activities of emerging phenolic compounds.

The screening of compounds with endocrine disrupting effects has been attracting increasing attention due to the continuous release of emerging chemicals into the environment. Testing the (ant)agonistic activities of these chemicals on the retinoic acid receptor α (RARα), a vital nuclear receptor, is necessary to explain their perturbation in the endocrine system in vivo. In the present study, MCF-7 breast carcinoma cells were transiently transfected with a RARα expression vector (pEF1α-RARα-RFP) and a reporter vector containing a retinoic acid reaction element (pRARE-TA-Luc). Under optimized conditions, the performance of the newly constructed system was evaluated for its feasibility in screening the (ant)agonistic effects of emerging phenolic compounds on RARα. The results showed that this transient transfection cell model responded well to stimulation with (ant)agonists of RARα, and the EC50 and IC50 values were 0.87 nM and 2.67 µM for AM580 and Ro41-5253, respectively. Its application in testing several emerging phenolic compounds revealed that triclosan (TCS) and tetrabromobisphenol A (TBBPA) exerted notable RARα antagonistic activities. This newly developed bioassay based on MCF-7 is promising in identifying the agonistic or antagonistic activities of xenobiotics on RARα and has good potential for studying RARα signaling-involved toxicological effects of emerging chemicals of concern.

Global Identification and Systematic Analysis of Lysine Malonylation in Maize (Zea mays L.).

Lysine malonylation is a kind of post-translational modifications (PTMs) discovered in recent years, which plays an important regulatory role in plants. Maize (Zea mays L.) is a major global cereal crop. Immunoblotting revealed that maize was rich in malonylated proteins. We therefore performed a qualitative malonylome analysis to globally identify malonylated proteins in maize. In total, 1,722 uniquely malonylated lysine residues were obtained in 810 proteins. The modified proteins were involved in various biological processes such as photosynthesis, ribosome and oxidative phosphorylation. Notably, a large proportion of the modified proteins (45%) were located in chloroplast. Further functional analysis revealed that 30 proteins in photosynthesis and 15 key enzymes in the Calvin cycle were malonylated, suggesting an indispensable regulatory role of malonylation in photosynthesis and carbon fixation. This work represents the first comprehensive survey of malonylome in maize and provides an important resource for exploring the function of lysine malonylation in physiological regulation of maize.

Tebuconazole induces liver injury coupled with ROS-mediated hepatic metabolism disorder.

Tebuconazole, the most widely used fungicide, is reported to cause various environmental problems and have serious health risks in humans. Despite numerous advances in toxicity studies, its internal metabolic process and the underlying mechanisms have not been systemically studied. The present study administered low doses (0.02 g/kg bw and 0.06 g/kg bw) of tebuconazole to C57BL/6 mice in vivo. The high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated to analyze the tebuconazole in different organs, and our data revealed that tebuconazole mainly accumulated in the liver and that histopathological damage were exhibited in this organ. Tebuconazole significantly dysregulated phase Ⅰ- and phase II-metabolizing enzymes, ATP-binding cassette (ABC) efflux transporters (Abcc2 and Abcc3) and fatty acid metabolism-related genes (Cdkn1a and Fasn), thereby directly causing liver hypertrophy and steatosis. Importantly, the excessive induction of reactive oxygen species (ROS) and oxidative stress partially accounted for the metabolic abnormalities mediated by tebuconazole. Moreover, these alterations were related to the abnormal transcriptional levels of peroxisome proliferator-activated receptor α (PPAR-α) and liver x receptor α (LXR-α), which were predicted to bind to tebuconazole via hydrogen bonding interactions. The current findings provide new insight into the molecular mechanisms of metabolic abnormalities induced by tebuconazole at low concentration, and are conducive to a better understanding of the environmental risk posed by this fungicide.

Nanoscale cobalt-based metal-organic framework impairs learning and memory ability without noticeable general toxicity: First in vivo evidence.

Metal-organic frameworks (MOFs) exhibit broad potential applications in the environmental, biomedical, catalyst, and energy fields. However, the currently existing data hardly shed light on their health risks before the MOFs' large-scale usage. In this context, we exploratively investigated the in vivo fate and effect of one representative cobalt-based zeolitic imidazolate framework (ZIF-67) at the nano- (60 nm) and submicron- (890 nm) scales. Different from submicron-scale ZIF-67 showing better biosafety, nanoscale particles manifested a neurodegenerative risk at the dose of no general toxicity, evidenced by the impairment of learning and memory ability and disordered function of the neuropeptide signaling pathway in a rat model. The involvement of oxidative damage and inflammatory processes in the neurotoxicity induced by ZIF-67 was discussed as well. These findings not only provide a wake-up call for the prudent applications of MOFs but also provide insight into the better design and safer use of MOFs for broader applications.

Graphene Quantum Dots Disrupt Embryonic Stem Cell Differentiation by Interfering with the Methylation Level of Sox2.

The tremendous potential for graphene quantum dots (GQDs) in biomedical applications has led to growing concerns of their health risks in human beings. However, present studies mainly focused on oxidative stress, apoptosis, and other general toxicity effects; the knowledge on the developmental toxicity and the related regulatory mechanisms is still far from sufficient. Our study revealed the development retardation of mouse embryonic stem cells (mESCs) caused by GQDs with a novel DNA methylation epigenetic mechanism. Specifically, GQDs were internalized into cells mainly via energy-dependent endocytosis, and a significant fraction of internalized GQDs remained in the cells even after a 48-h clearance period. Albeit with unobservable cytotoxicity or any influences on cell pluripotency, significant retardation was found in the in vitro differentiation of the mESCs into embryoid bodies (EBs) with the upregulation of Sox2 levels in GQD pretreatment groups. Importantly, this effect could be contributed by GQD-induced inhibition in CpG methylation of Sox2 through altering methyltransferase and demethyltransferase transcriptional expressions, and the demethyltransferase inhibitor, bobcat339 hydrochloride, reduced GQD-induced upregulation of Sox2. The current study first demonstrated that GQDs compromised the differentiation program of the mESCs, potentially causing development retardation. Exposure to this nanomaterial during gestation or early developmental period would cause adverse health risks and is worthy of more attention.

Gold nanoparticles change small extracellular vesicle attributes of mouse embryonic stem cells.

Gold nanoparticles (AuNPs) have attracted considerable interest in suppressing tumor cell migration, while small extracellular vesicles (sEVs) play an essential role in tumor metastasis by shaping the tumor microenvironment. Understanding how AuNPs alter sEV attributes is critical in antitumor medication design. In this study, mouse embryonic stem cells (mESCs) were treated with three sizes of AuNPs (i.e. 5 nm AuNPs, 20 nm AuNPs, and 80 nm AuNPs) to obtain sEVs (i.e. sEV-5, sEV-20, and sEV-80), which were characterized from the biophysical and proteomic aspects. When compared with the control (sEV-ctrl), sEV-5 possessed relatively higher rigidity, and a differentially expressed protein profile. It attenuated 4T1 tumor cell proliferation and migration through inhibiting cofilin expression and extracellular regulated protein kinase (Erk) phosphorylation, which was opposite to the effect induced by sEV-ctrl. In contrast, sEV-20 and sEV-80 had negligible effects. This study revealed for the first time that AuNP-5 exposure changed the biophysical properties and cellular functions of mESC-derived sEVs, providing a promising strategy for designing AuNP-based antitumor medication.

Sex difference in bronchopulmonary dysplasia of offspring in response to maternal PM2.5 exposure.

The adverse effects of fine particulate matters (PM2.5) on respiratory diseases start in utero. In order to investigate whether maternal PM2.5 exposure could lead to bronchopulmonary dysplasia (BPD) in offspring, PM2.5 was collected in Taiyuan, Shanxi, China during the annual heating period. Mice were mated and gestation day 0 (GD0) was considered the day on which a vaginal plug was observed. The plug-positive mice received 3 mg/kg b.w. PM2.5 by oropharyngeal aspiration every other day starting on GD0 and throughout the gestation period. Offspring were sacrificed at postnatal days (PNDs) 1, 7, 14 and 21. We assessed some typical BPD-like symptoms in offspring. The results showed that maternal PM2.5 exposure caused low birth weight, hypoalveolarization, decreased angiogenesis, suppressed production of secretory and surfactant proteins, and increased inflammation in the lungs of male offspring. However, maternal PM2.5 exposure induced only hypoalveolarization and inflammation in the lungs of female offspring. Furthermore, these alterations were reversed during postnatal development. Our results demonstrated that maternal exposure to PM2.5 caused reversible BPD-related consequences in offspring, and male offspring were more sensitive than females. However, these alterations were reversed during postnatal development.

Investigating photo-driven arsenics' behavior and their glucose metabolite toxicity by the typical metallic oxides in ambient PM2.5.

It is essential and challenged to understand the atmospheric arsenic pollution because it is much more complicated than in water and top-soil. Herein the different behavior of arsenic species firstly were discovered within the ambient PM2.5 collected during daytime and nighttime, winter and summer. The diurnal variation of arsenic species in PMs is significantly correlated with the presence of metallic oxides, specifically, ferrous, titanium and zinc oxides, which might play a key role in the process of the photo-oxidation of As(III) to As(V) with the meteorological parameters and regional factors excluded. Subsequently, the photo conversion of arsenite was detected on metal-loaded glass-fiber filters under visible light. The photo-generated superoxide radical was found to be predominantly responsible for the oxidation of As(III). In order to reveal toxicity differences induced by oxidation As(III), HepG2 cells were exposed to various arsenic mixture solution. We found that the antioxidant enzyme activities suppressed with increasing the As(III)/As(V) ratio in total, followed by the accumulation of intracellular ROS level. The glucose consumption and glycogen content also displayed an obvious reduction in insulin-stimulated cells. Compared to the expression levels of IRS-1, AKT and GLUT4, GLUT2 might be more vulnerable to arsenic exposure and lead to the abnormalities of glucose metabolism in HepG2 cells. Taken together, these findings clarify that the health risk posed by inhalation exposure to As-pollution air might be alleviated owing to the photo-driven conversion in presence of metal oxides.

PM2.5 exposure induces age-dependent hepatic lipid metabolism disorder in female mice.

Particulate matter exposure has been described to elevate the risk of lung and cardiovascular diseases. An increasing number of recent studies have indicated positive correlations between PM2.5 (the fraction of airborne particles with an aerodynamic diameter less than 2.5 µm) exposure and the risk of liver diseases. However, research on the effects of PM2.5 exposure on liver fat synthesis, secretion, and clearance mechanisms under normal diet conditions is limited, and whether these effects are age-dependent is largely unknown. Female C57BL/6 mice at different ages (4 weeks (4 w), 4 months (4 m), and 10 months (10 m)) were treated with 3 mg/kg body weight of PM2.5 every other day for 4 weeks. Subsequently, the ultrastructural changes of liver, the expression of genes involved in oxidative damage and lipid metabolism in the liver were examined. Observation of hepatic ultrastructure showed more and larger lipid droplets in the livers of 4-week-old and 10-month-old mice exposed to PM2.5. Further analysis showed that PM2.5 exposure increased the expression of genes related to lipid synthesis, but decreased the expression of genes involved in lipid transport and catabolism in the livers of 10-month-old mice. Our findings suggest that exposure to PM2.5 disrupts the normal metabolism of liver lipids and induces lipid accumulation in the liver of female mice in an age-dependent manner, with older mice being more susceptible to PM2.5.

Assessment of the carcinogenic effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin using mouse embryonic stem cells to form teratoma in vivo.

As the most toxic dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has gained lots of concerns, due to its diverse deleterious effects. However, the knowledge on carcinogenic risk of TCDD during early stage of development remains scarce. The in vivo teratoma formation model based on the transplantation of embryonic stem cells (ESCs) in immunodeficient mice is appealing for studying pluripotency and tumorigenicity in developmental biology, and also shows promise in environmental toxicology, especially in carcinogenesis researches. In this study, the malignant transformation of mouse embryonic stem cells (mESCs) pretreated with TCDD was investigated during their in vivo differentiation using teratoma formation model. Based on characterization of the pluripotency and differentiation capabilities of mESCs, evil changes in teratomas derived from TCDD-exposed mESCs were systematically studied. The results showed that TCDD significantly up-regulated CYP1A1 transcriptional levels in mESCs, elevated the incidence of malignant change in mESC-derived teratomas, and caused indefinite proliferation capabilities in sequential cultures of tumor tissues. The findings suggested that TCDD could exert carcinogenic effect on mESCs during their differentiation into teratoma in vivo, and more attention should be paid to the adverse health effects of this chemical during gestation or early developmental period.

Ambient fine particulate matter exposure induces cardiac functional injury and metabolite alterations in middle-aged female mice.

Plenty of epidemiological studies have shown that exposure to ambient particulate matter (PM2.5) is linked to cardiovascular diseases (CVDs) in older even in middle-aged populations; however, experimental evidence through intuitive metabolic analysis to confirm the age susceptibility and explain the related molecular mechanism of PM2.5-induced cardiotoxicity is relatively rare. In the present study, C57BL/6 mice (adult (4-month) and middle-aged (10-month)) were given 3 mg/kg PM2.5 every other day by oropharyngeal aspiration for 4 weeks, and then, body and cardiac parameter, containing weight data, cardiac function, ultrastructure, metabolic analysis, and molecular detection were conducted to investigate the PM2.5-induced cardiotoxicity. The results indicated that middle-aged mice were more susceptible to PM2.5, displaying slow cardiac growth, cardiac dysfunction, abnormal mitochondrial structure and function, and cardiac metabolic disorders. The altered metabolites were enriched in carbohydrate metabolism, fatty acid metabolism, amino acid metabolism, nucleotide metabolism and nicotinate and nicotinamide metabolism. In conclusion, we speculated that the cardiac metabolic disorders may be important factors in PM2.5-induced cardiac dysfunction and mitochondrial structure destruction in middle-aged mice, providing a new direction for the study of the association between PM2.5 and CVDs.

Histone modification in the lung injury and recovery of mice in response to PM2.5 exposure.

Epidemiological and experimental studies have progressively provided a better knowledge of the underlying mechanisms by which fine particulate matter (PM2.5) exerts its harmful health effects. However, limited studies focused on the effect and following recovery after the particulate exposure ended. In this study, we determined PM2.5 exposure-caused effects on the lung and their recovery in mice after terminating aspiration, and clarified the possible molecular modification. The results revealed that PM2.5 exposure for 4 weeks significantly decreased the lung function, and the changes returned to normal levels after 1-week recovery. However, we observed persistent particle alveolar load following 2-week recovery. Interestingly, the alterations of H3K27ac expression and related enzyme activities mimicked the changes of respiratory function during the process, and chromatin immunoprecipitation-seqences (ChIP-seq) suggested that these PM2.5-associated differential H3K27ac markers participated in immune responses and chemokine signaling pathway with stat2 and bcar1 being two important genes. Consistently, the expression of pro-inflammatory cytokines and chemokines elevated after PM2.5 exposure for 4-week, and reversed to normal levels following 2-week recovery. The study highlighted that PM2.5 aspiration caused histone modification associated lung dysfunction and inflammation, and the action restored after exposure ending and 2-week recovery. Also, persistent particle alveolar load might be a long-term potential risk for lung diseases.

Comprehensive hippocampal metabolite responses to PM2.5 in young mice.

Fine particulate matter (PM2.5) exposure alters brain development, clinical cognition and behavior in childhood. Previous studies of this subject have mainly been epidemiological investigations or analyses of gene and protein levels; however, gas chromatography-mass spectrometry (GC-MS)-based metabolic profiling, which will help clarify the molecular mechanisms of susceptibility in PM2.5-induced neurotoxicity, is lacking. In the present study, C57BL/6 mice at different ages (4 weeks, 4 months and 10 months) received oropharyngeal aspiration of PM2.5 (3 mg/kg) every other day for 4 weeks. The Morris water maze showed that PM2.5 exposure caused deterioration of spatial learning and memory in young (4 week old) mice. In addition, the levels of several metabolites belonging to different metabolite classes were significantly changed by PM2.5 exposure in 4-week-old mice. Based on metabolic pathway analysis, we speculated that the decline in spatial learning and memory due to PM2.5 exposure may be directly or indirectly associated with hippocampal region-specific metabolic alterations involving energy metabolism (citric acid, succinic acid, malic acid, maltose and creatinine); cholesterol metabolism (desmosterol, lanosterol and campesterol); arachidonic acid metabolism (methyl arachidonic acid, nonanoic acid and linoleic acid); inositol phosphate metabolism (myo-inositol, myo-inositol-1-phosphate and methyl-phosphate) and aspartic acid metabolism (aspartic acid, asparagine and homoserine).