Toxicity Of Dibutyl Phthalate Research Articles

Toxicity of Dibutyl phthalate (DBP) toward isolated human blood lymphocytes: Apoptosis initiated from intracellular calcium enhancement and mitochondrial/lysosomal cross talk

Dibutyl phthalate (DBP) is a phthalate ester with wide application in industrial products, so human exposure can happen in workplaces and environment. Conflicting results have been acquired in researches which measured the influences of phthalates contact on immune responses in laboratory animals. Nevertheless, the straight influence of DBP on human lymphocytes and entire mechanisms of its effect against these cells continue to be unexplored. The major purpose of present research was to evaluate the mechanisms which lead to the DBP toxicity on human lymphocytes using accelerated cytotoxicity mechanisms screening (ACMS) technique. Cell viability was determined following12h incubation of lymphocytes with 0.05–1 mM DBP, and mechanistic parameters were assessed after 2, 4 and 6 h of lymphocyte treatment with ½ the IC5012h (0.3 mM), the IC5012h (0.6 mM) and twice the IC5012h (1.2 mM) of DBP. The IC5012 h of a chemical/toxicant is defined as concentration that kills 50 % of cells after 12 h of exposure. The results indicate that DBP exerts toxic effects on isolated human lymphocytes, probably through mitochondrial and lysosomal damage induced by glutathione depletion and oxidative stress. In this study, suppression of cytokines (IL2, INF-gamma and TNF-alpha) production and increase in intracellular calcium were also related to DBP induced lymphocyte toxicity.

Toxic mechanism in Daphnia magna due to phthalic acid esters and CuO nanoparticles co-exposure: The insight of physiological, microbiomic and metabolomic profiles

Various phthalic acid esters (PAEs) such as dibutyl phthalate (DBP) and butyl benzyl phthalate (BBP) co-exist with nanopollutants in aquatic environment. In this study, Daphnia magna was exposed to nano-CuO and DBP or BBP at environmental relevant concentrations for 21-days to investigate these combined toxic effects. Acute EC50 values (48 h) of nano-CuO, DBP, and BBP were 12.572 mg/L, 8.978 mg/L, and 4.785 mg/L, respectively. Results showed that co-exposure with nano-CuO (500 μg/L) for 21 days significantly enhanced the toxicity of DBP (100 μg/L) and BBP (100 μg/L) to Daphnia magna by 18.37% and 18.11%, respectively. The activities of superoxide dismutase, catalase, and glutathione S-transferase were enhanced by 10.95% and 14.07%, 25.63% and 25.91%, and 39.93% and 35.01% in nano-CuO+DBP and nano-CuO+BBP treatments as compared to the individual exposure groups, verifying that antioxidative defense responses were activated. Furthermore, the co-exposure of nano-CuO and PAEs decreased the population richness and diversity microbiota, and changed the microbial community composition in Daphnia magna. Metabolomic analysis elucidated that nano-CuO + PAEs exposure induced stronger disturbance on metabolic network and molecular function, including amino acid, nucleotides, and lipid metabolism-related metabolic pathways, as comparison to PAEs single exposure treatments. In summary, the integration of physiological, microflora, and untargeted metabolomics analysis offers a fresh perspective into the potential ecological risk associated with nanopollutants and phthalate pollution in aquatic ecosystems.

Multispectral method combined with molecular modelling to investigate the binding mechanisms of DBP and DIBP on pepsin

The binding mechanisms of dibutyl phthalate (DBP) and diisobutyl phthalate (DIBP) with pepsin (PEP) were investigated by multispectral method and computational techniques (molecular docking, molecular dynamics, surface electrostatic potential), as well as cytotoxicity assay. Fluorescence studies revealed that both DBP and DIBP had a static bursting effect on PEP, forming a stable complex. Furthermore, the formation of PEP-DBP/DIBP complexes was observed to occur spontaneously (ΔG < 0), driven by the interactions of hydrogen bonding and van der Waals forces. PEP exhibits a higher binding constant Ka of DBP (3.30 × 104 L·mol−1) compared to that of DIBP (2.37 × 104 L·mol−1). The higher stability of PEP-DBP complexes can be attributed to the different branching structures of DBP and DIBP, this observation was further supported by the larger negative surface area of DBP (125.90 Å2) in the ESP analysis, and were aligned with the MD simulations results of more stable RMSD and lower Rg value in presence of DBP. Computational analysis revealed that both DBP and DIBP exhibited binding interactions with the fluorescent groups (Tyr 75, Phe 111, and Phe 117) by van der Waals forces. DBP/DIBP binds to the Asp 32 and Asp 215 motifs of the catalytic activity center of PEP. Combined with the lower Rg values, CD tests suggested that the addition of DBP/DIBP caused the rise of α-Helix content thereby compacted the conformation structure of PEP into a compact shape. In addition, DBP generated more hydrogen bonds with PEP compared to DIBP, also as evidence to the higher stability of EPE-DBP. Finally, the IC50 of DBP and DIBP on Caco-2 cells were determined to be 5.16 × 10-4 mol·L-1 and 1.37 × 10-3 mol·L-1, respectively confirming the higher level of toxicity of DBP. The present study elucidates the effect and binding mechanism of DBP/DIBP on PEP at the molecular level, providing fundamental data for the impact and potential risk assessment of PAEs on the human digestive system.

Multi-walled carbon nanotubes enhance the toxicity effects of dibutyl phthalate on early life stages of zebrafish (Danio rerio): Research in physiological, biochemical and molecular aspects

Phthalate esters (PAEs) are widely used as plasticizers. PAEs are ubiquitous in natural water bodies, with dibutyl phthalate (DBP) being one of the most common PAEs. DBP is prone to leaching or migration into the environment, posing serious health and environmental risks. Carbon nanotubes (CNTs) have been widely used in various fields with the rapid development of nanotechnology. CNTs could alter the environmental behavior and toxicity of co-existing pollutants. CNTs have been shown to rapidly adsorb PEAs. However, current knowledge about the effects of CNTs on DBP toxicity is limited. Here we show that the toxic effects of single and combined exposure to DBP (0.1, 0.5, 1.0 mg/L) and different CNTs (MWCNTs/MWCNTs-COOH, 0.5 mg/L) on the early growth stage of zebrafish. The results suggested that a significant increase in heart rate and heart malformation rate was observed after co-exposure of DBP and MWCNTs/MWCNTs-COOH (p < 0.05). Furthermore, combined exposure increased antioxidant enzyme activity during early developmental stages in zebrafish (p < 0.05). The qRT-PCR results revealed that DBP and MWCNTs/MWCNTs-COOH co-exposure significantly interfered with the expression of genes related to oxidative stress, energy metabolism, development of cardiac function, and apoptosis (p < 0.05). In addition, for oxidative stress and cardiotoxicity, MWCNTs/MWCNTs-COOH aggravated the toxic effects of 0.5 mg/L DBP on embryos/larvae. The metabolomics results showed that co-exposure mitigated the disturbance of amino acid metabolism mediated by single DBP exposure. In general, MWCNTs/MWCNTs-COOH increased the impact of DBP in the early developmental stages of zebrafish. This study provides new insights into the toxicology of early developmental stages of aquatic organisms exposed to co-exist pollutants of DBP and CNTs.

Di-butyl phthalate disrupts muscle, motor and sensory neuron development in embryonic zebrafish

Phthalates are added to plastics to enhance its flexibility, durability and transparency. Phthalates are not covalently bound to plastics and leach into the environment. Phthalates are now pervasive and ubiquitously present in the atmosphere, soil and sediment, surface and wastewater. Phthalates are known endocrine disruptors and their effects on male and female reproduction are well noted. However, studies on the developmental effects of di-butyl phthalate (DBP) are limited. Here we investigated the developmental toxicity of DBP on motor and sensory neuron populations and the muscle structures motor neurons innervate using the zebrafish vertebrate model system. We investigated these effects during the time window of development spanning the period where embryonic patterning determines adult structures. We found that treatment with 2.5 μM DBP from 6 h post-fertilization (hpf) until 72hpf induces loss and disorganization of primary motor neuron innervation of the somatic tissue with concomitant disruptions to muscle fiber organization. Furthermore, we found disruptions to sensory motor neuron development including defects in dorsal root ganglion and their peripherally extending axons. Rohon-Beard sensory neurons were also disrupted showing loss of the bilateral soma positioning along the length of the spinal cord and their afferent axonal projections to the epithelium. Thus, we concluded that DBP is toxic to developing motor and sensory neurons during embryonic development.

Ferroptosis participates in dibutyl phthalate-aggravated allergic asthma in ovalbumin-sensitized mice

Dibutyl phthalate (DBP), used as a plasticizer, is of wide concern as an environmental pollutant since it has certain immunotoxicity. Although there is growing evidence supporting a link between DBP exposure and allergic airway inflammation, there is less information concerned with whether the ferroptosis pathway is involved in DBP-aggravated allergic asthma in ovalbumin (OVA)-sensitized mice. This study aimed to investigate the role and underlying mechanisms of ferroptosis in DBP-exposed allergic asthmatic mice. Balb/c mice were orally exposed to 40 mg/kg−1 DBP for 28 days, followed by sensitization with OVA and seven consecutive challenges with nebulized OVA. We analyzed airway hyperresponsiveness (AHR), immunoglobulins, inflammation and pulmonary histopathology, to investigate whether DBP exacerbates allergic asthma in OVA-induced mice. We also measured the biomarkers of ferroptosis (Fe2+, GPX4, PTGS2), proteins related to the ferroptosis pathway (VEGF, IL-33, HMGB1, SLC7A11, ALOX15, PEBP1), and indices of lipid peroxidation (ROS, Lipid ROS, GSH, MDA, 4-HNE), to explore the role of ferroptosis in DBP+OVA mice. Finally, we used ferrostatin-1 (Fer-1) as an antagonist against the harmful effects of DBP. The results showed that, DBP+OVA mice had a significant increase in AHR, airway wall remodeling and airway inflammation. Further, we showed that DBP aggravated allergic asthma via ferroptosis and lipid peroxidation, and that Fer-1 inhibited ferroptosis and alleviated the pulmonary toxicity of DBP. These results suggest that ferroptosis participates in the exacerbation of allergic asthma resulting from oral exposure to DBP, highlighting a novel pathway for the connection between DBP and allergic asthma.

Insight into the health risk implicated in mitochondrial toxicity of dibutyl phthalate exposure on zebrafish (Danio rerio) cells

Dibutyl phthalate (DBP) is commonly applied plasticizer in plastic products such as face masks, easily leaches or migrates into environment and its widespread contamination posed profound health risks. Further concerns rise regarding to the toxicity of DBP at subcellular level, while little is known about the ranging effects on mitochondrial susceptibility. Present study investigated the mitochondrial impairments with implicated cell death upon DBP exposure on zebrafish cells. Elevated mitochondrial oxidative stress reduced its membrane potential and count, enhanced fragmentation, and impaired ultrastructure that showed smaller size and cristae rupture. Afterwards, the critical function of ATP synthesis was damaged and the stabilized binding capacity between DBP with mitochondrial respiratory complexes was simulated by the molecular docking. And the top pathways enrichment of mitochondrion and metabolism by transcriptome analyses verified the mitochondrial dysfunction that indicated the human diseases risks. The mitochondrial DNA (mtDNA) replication and transcription with DNA methylation modifications were also disrupted, reflecting the genotoxicity on mtDNA. Moreover, the activated autophagy and apoptosis underlying mitochondrial susceptibility integrated into cellular homeostasis changes. These findings provide the first systemic evidence broadening and illustrating the mitochondrial toxicity of DBP exposure on zebrafish model that raise concern on phthalates contamination and ecotoxicological evaluation.

Toxicity of dibutyl phthalate to pakchoi (Brassica campestris L.): Evaluation through different levels of biological organization

Dibutyl phthalate (DBP) is a typical persistent organic pollutant with a high load in the agricultural soils of vegetable crops. Currently, studies on the toxicity of DBP in vegetable crops are limited. Therefore, in this study, pakchoi (Brassica campestris L.), a typical vegetable crop, was used to evaluate the toxic effects of DBP. Pakchoi was exposed to DBP for 24 d at three doses (2, 20, and 200 mg/kg), and the phenotypic, biochemical, and molecular indicators were determined. The results revealed that DBP could reduce the emergence of pakchoi and inhibit plant height, root length, fresh weight, and leaf area. At the biochemical level, DBP exposure could reduce the content of three typical photosynthetic pigments (chlorophyll a and b and carotenoids). The effects of DBP exposure on the quality of pakchoi were primarily through reduced soluble sugar and increased proline contents. In addition, O2·− and H2O2 levels increased after DBP stress, and the corresponding antioxidant enzymes (SOD, POD, and CAT) were activated to resist oxidative damage. The dose- and time-dependent toxicities of DBP to pakchoi were demonstrated using an integrated biological response index. Finally, the molecular-level results on Day 24 showed that the three antioxidant enzyme genes (sod, pod, and cat) were significantly downregulated, and the antioxidant enzyme genes were more sensitive biomarkers than the enzyme activities. However, the expression level of enzyme genes was opposite to that of enzyme activity (SOD and POD); thus, DBP might directly interact with these enzymes. Molecular docking showed that DBP could stably bind near the SOD/POD active center through intermolecular interaction forces. This study provides essential information on the risk of DBP toxicity to vegetable crops.

Accumulation, detoxification, and toxicity of dibutyl phthalate in the swimming crab

Dibutyl phthalate (DBP) is one of the most commonly used and toxic phthalate esters and has a variety of harmful effects on aquatic animals. However, there is still a lack of knowledge on the accumulation, detoxification, and toxicity of DBP in aquatic animals. In this study, we chose the swimming crab Portunus trituberculatus, an ecologically and economically important species, as the model and investigated the metabolism of DBP and its effects on the detoxification, antioxidation, survival and growth of the crab juveniles to better understand DBP-triggered molecular response over different time courses. As a result, DBP could be accumulated in the swimming crab in a concentration-dependent manner and metabolized to monobutyl phthalate (MBP) and phthalic acid (PA) through de-esterification. DBP exposure induced the different responses of three cytochrome P450 members and antioxidant enzyme genes, enhanced gene transcript and protein levels of glutathione-S-transferase and two heat stress proteins and malondialdehyde accumulation, decreased glutathione level, and inhibited antioxidant enzyme activities. Further, no significant effect of DBP was observed in crab survival, size, and weight but there was molting retardation. Therefore, DBP induced strong detoxification and antioxidative defense mechanisms to overcome detrimental effects of DBP on the swimming crab juveniles despite a molting retardation as a trade-off in fitness costs. The prevalent coexistence of DBP with MBP and PA during the whole exposure period is raising concerns on the combined action and ecological risk to aquatic animals.

Dibutyl phthalate induced oxidative stress and genotoxicity on adult zebrafish (Danio rerio) brain

Dibutyl phthalate (DBP) is one of the most widely used plasticizers with a high concentration in the water. Although the toxicity of DBP on aquatic organisms has become a significant concern in recent years, the effects of DBP on zebrafish (Danio rerio) brain is poorly understood. This study investigated the toxic effects of DBP exposure for 7, 14, 21 and 28 days on zebrafish brain. The results showed that DBP significantly stimulated SOD and CAT activities, increasing MDA and 8-OHdG contents. On the 28th day, the AChE inhibition rates in 0.08, 0.4, 2 mg·L−1 treatment were 13.4%, 11.9%, 14.7%. The trend of Cu/Zn-sod gene variation was consistent with SOD activity, showing “inhibition-activation-inhibition”. The expression of apoptotic genes (caspase-3, p53) showed “inhibition-activation-inhibition”. The integrated biomarker response (IBR) results showed that the IBR values were 4.37, 7.18 and 9.63 in 0.08, 0.4 and 2 mg·L−1 group on the 28th day, presenting a “dose-response” relationship. These findings confirmed that low concentration of DBP induced oxidative damage and genotoxicity in zebrafish brain, which provided an effective toxicological basis for phthalate pollution. Based on above studies, it is of great significance for assessing the harmful effects of DBP with low concentration on aquatic organisms.

Di-butyl phthalate (DBP) induces defects during embryonic eye development in zebrafish.

Di-butyl phthalate (DBP) is a phthalate ester (PAEs) added during the manufacturing of plastics to make them stronger, yet more pliable. DBP is noncovalently bound to plastics resulting in leaching into the environment. Concerning concentrations of DBP have been noted in surface and groundwater, aquatic ecosystems, soil and atmospheric environments globally. Global production of phthalates and thus concomitant exposure has increased over the years making studies on the ecological and environmental safety needed. Most of the literature on DBP focuses on the endocrine disrupting properties of phthalate esters, but the developmental toxicity of DBP is an understudied area. Here, we treat gastrula staged zebrafish embryos with environmentally relevant concentrations of DBP (2.5 µM). We find defects in eye development at 96 h post fertilization including a decrease in the size of the lens and retina in DBP-treated embryos. Defects in eye vascularization as well as loss of the optic nerve and optic tectum were also noted. Here we conclude that exposure to environmentally relevant doses of DBP during early embryonic development is toxic to eye development.

Key role of exopolysaccharide on di-butyl phthalate adsorbing by Lactobacillus plantarum CGMCC18980

Plasticizers belong to hormone-like substances existing widely in the environment. According to the Environmental Protection Agency of China, they are considered to be the fourth class of toxic chemicals due to their harmful effects on normal endocrine system in human bodies. In the recent published work of our lab, Lactobacillus plantarum CGMCC18980 (strain P1) could reduce the toxicity of di-butyl phthalate (DBP) in rats effectively. The purpose of this study is to further explore the adsorption mechanism of di-butyl phthalate to L. plantarum CGMCC18980, based on optimizing the adsorption conditions. As a consequence, the adsorption effect of L. plantarum CGMCC18980 attributed to relationships between exopolysaccharide, membrane protein, and the cell wall. Experimental results demonstrated that exopolysaccharide and the cell wall were devoted to DBP binding. An obvious adsorption layer was observed outside of L. plantarum CGMCC18980 through scanning electron microscope (SEM) and transmission electron microscope (TEM). The Fourier transform infrared spectroscopy (FTIR) results showed that the functional groups involved in adsorption were mainly C=O, C-N, and C-O, which related to lipids and polysaccharides. Zeta potential analysis indicated that DBP adsorption had no significant relationship with surface charge. These results revealed that exopolysaccharide may be the key factor of strain CGMCC18980 in DBP adsorption.Key points• Lactobacillus plantarum CGMCC18980 has the ability to adsorb di-butyl phthalate, reaching to 58.63%.• Exopolysaccharide is considered to play a key role in adsorption process.• Membrane protein, cell wall, and surface charge do not contribute to adsorption.

The overview of current evidence on the reproductive toxicity of dibutyl phthalate.

Over the past years, many legitimate concerns have been raised about the effects of dibutyl phthalate (DBP) as an endocrine disruptor, especially on reproduction. The aim of this publication is to critically review the literature related to the developmental and reproductive toxicity of DBP in animals. Several electronic databases were systematically searched until 2019. Studies were qualified for the review if they: linked exposure to DPB with reproduction, were published in English after 1990, and were conducted on animals. In the studies of the testicular effects of DBP on experimental animals, the most common effects of exposure included reduced fertility, atrophic changes in male gonads, degenerative changes in the epididymis, as well as a reduction in sperm count and motility, cryptorchidism, hypospadias, poor sperm quality and other genital defects (decreased testicular weight, delayed spermatogenesis, Leydig cell aggregation, impaired Sertoli cell maturation, and significant inhibitions of testicular enzymes). The embryotoxic effects of DBP on laboratory animals included mainly an increase in fetal resorption and a decrease in live births. The teratogenic effects of DBP also manifest as skeletal malformations in fetuses, malformations of male gonads and other genital effects. On the basis of the literature data, it is clearly demonstrated that DBP shows anti-androgenic effects; however, there are also reports confirming its weak estrogenic effect. Additionally, lower doses cause more adverse effects than the highest dose, which is an important fact because of the widespread environmental exposure to DBP. The studies clearly confirm that DBP is an endocrine disruptor. Int J Occup Med Environ Health. 2021;34(1):15-37.

Biodegradation of dibutyl phthalate by a novel endophytic Bacillus subtilis strain HB-T2 under in-vitro and in-vivo conditions

ABSTRACT The environmental prevalence and potential toxicity of dibutyl phthalate (DBP) motivate the attempt to develop feasible strategies to deal with DBP contamination. In this study, a strain of endphytic bacteria HB-T2 was isolated from sorrel roots and identified as Bacillus sp. by analysing its morphology, physiology, biochemistry and 16S rDNA sequence. The degradation efficiency of DBP by HB-T2 was almost identical under the temperature of 30∼40°C, but was significantly enhanced as the culture pH and inoculum size increases from 6.0 to 8.0, and 1% to 5% respectively. The degradation kinetics of DBP could be well described by the first-order kinetic model, with the degradation half-life ranging from 1.59 to 7.61 h when the initial concentrations of DBP were in the range of 5–20 mg/L. LC-MS analysis of the culture samples taken at varying intervals revealed monobutyl phthalate, phthalic acid and protocatechuic acid as the major metabolic intermediates during the degradation process. HB-T2 exhibited an excellent capability to degrade a wide range of phthalate esters (PAEs), especially butyl benzyl phthalate (BBP), dipentyl phthalate (DPP), and diisobutyl phthalate (DIBP). Inoculation of HB-T2 into Chinese cabbage (Brassica chinensis L.) growing in DBP-contaminated soils could significantly reduce the DBP levels in plant tissues and relieve the phytotoxic effects of DBP. Results of this study highlighted the great potential of this novel endophytic Bacillus subtilis strain HB-T2 for bioremediation of PAEs contamination and improvement of agricultural product safety by reducing PAEs accumulation in edible crops.

Dibutyl phthalate-induced oxidative stress, inflammation and apoptosis in grass carp hepatocytes and the therapeutic use of taxifolin

The widespread use of plastic products has led to the widespread presence of plasticizers in the environment. As a common environmental pollutant, research on plasticizer toxicity is insufficient in fish cells. In particular, research on the toxicity of dibutyl phthalate (DBP) in grass carp hepatocyte lines is insufficient. To further explore these mechanisms, we treated grass carp hepatocytes with 300 μM DBP, a common plasticizer, for 24 h, and hepatocytes were also treated with 1 μM taxifolin (TAX), an antioxidant, for 24 h to study its antagonistic effect on DBP. After DBP exposure, oxidative stress levels and inflammation in hepatocytes increased, and the mRNA and protein expression of apoptosis-related markers increased significantly, leading to hepatocyte apoptosis. Moreover, AO/EB staining, Hoechst staining and flow cytometry also showed that the level of apoptotic cells increased after DBP exposure. Notably, both TAX pretreatment and TAX simultaneous treatment alleviated oxidative stress, increased inflammatory factor levels and apoptosis induced by DBP. In comparison, the effect of simultaneous TAX treatment was better than that of TAX pretreatment. Our results showed that TAX alleviates DBP-induced apoptosis in grass carp hepatocytes through oxidative stress and inflammation, and TAX pretreatment and simultaneous treatment exhibited specific effects. Specifically, simultaneous treatment had a better effect. Our study assessed the toxicity of DBP in grass carp hepatocytes and provided a theoretical and research basis for the in vivo study of animal models in the future. The innovation of this study involves the exploration of the interaction between DBP and TAX for the first time. This study may enrich knowledge regarding the theoretical mechanism of DBP toxicity in fish hepatocytes and propose methods address DBP toxicity.

Acute and chronic combined effect of polystyrene microplastics and dibutyl phthalate on the marine copepod Tigriopus japonicus

Dibutyl phthalate (DBP) is a commonly used additive in plastic products, so it may potentially coexist with microplastics (MPs) in marine environment. The ingestion of MPs might affect the accumulation of DBP in marine organisms. In this study, the marine copepod Tigriopus japonicus was applied to study the combined effect of DBP and polystyrene microplastics (mPS) on the copepod through both acute mortality tests and chronic reproduction tests. The LC50 of DBP was 1.23 mg L−1 (95% CI: 1.11–1.35 mg L−1), while exposure to mPS didn’t have significant lethal effect on the copepods. Adsorption to MPs led to decreased bioavailability of DBP, resulting in decreased toxicity of DBP. In contrast to the results of acute toxicity tests, DBP didn’t affect the reproduction of the copepods at lower exposure concentrations, while mPS reduced the number of nauplii and extended the time to hatch. Similar as acute toxicity tests, antagonistic interaction was observed for mPS and DBP in chronic reproduction tests, which might be attributed to promoted aggregation of mPS at presence of DBP. Overall, antagonistic toxicity effect between the two pollutants was observed for both acute and chronic tests, but the mechanisms of the interaction between DBP and mPS were different. Results of the present study highlighted the importance of long-term exposure when evaluating the toxic effect of MPs and their combined effect with other chemicals.

Involvement of dibutyl phthalate on male reproductive toxicity in the freshwater fish Pseudetroplus maculatus (Bloch, 1795)

ABSTRACT The testicular toxicity of dibutyl phthalate (DBP) was evaluated on the freshwater fish Pseudetroplus maculatus. The median lethal concentration of DBP for 96 h determined by probit analysis was 2 mg L−1, and its sublethal (0.2 mg L−1) exposure for 1, 3, 4, 7, and 15 d showed no change in the body weight but affected the reproductive parameters. The concentrations of alkali-labile phosphates and total protein in plasma and testis increased significantly (P < 0.05) with increase in the level of estradiol and decrease in testosterone, illustrating the adverse effect of DBP on male reproductive functions. Impairment in testicular steroidogenic enzymes and reduction in serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), and thyroid stimulating hormone (TSH) with increase in cortisol suggested reproductive dysfunction and stress response of the fish. Further, failure in the reproductive performance was evidenced by reduction in sperm motility, viability, and sperm count. Histological examination confirmed the absence of spermatozoa, completely disorganized spermatogonia, and spermatocytes in treatment groups, suggesting testicular toxicity of DBP, which may contribute to infertility and decline in fish population in the natural environment.

Toxicity of dibutyl phthalate in primary cultured rat hippocampal neurons and the toxicological mechanism

To investigate the neurotoxicity and toxicological mechanism of dibutyl phthalate (DBP) in primary cultured rat hippocampal neurons. Primary rat hippocampal neurons cultured for 4 days were exposed to 1 g/L DBP for 24, 48, or 96 h. Immunofluorescence assay and transmission electron microscopy (TEM) were used to observe the morphological changes of the axons and the ultrastructure of DBP-treated neurons. The action potential (AP) of the hippocampal neurons was measured with patch-clamp electrophysiology. CCK-8 assay was used to detect the viability of the hippocampal neurons, and Western blotting was performed to determine the mRNA and protein expressions of brain-derived neurotrophic factor (BDNF), neuropeptide Y (NPY) and estrogen receptor β (ERβ). High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS) was employed to detect the release of the neurotransmitter GABA. After exposure to DBP for 96 h, the cellular network of the hippocampal neurons became sparse, and the neurons showed significantly decreased axonal length (P < 0.01) and presented with round cell nuclei, chromatin aggregation and cytoplasmic vacuolization. Patch-clamp electrophysiology revealed depolarization drift and increased frequency of discharge in the exposed neurons (P < 0.01). The neurons with DBP exposure for 24, 48 and 96 h all showed significantly decreased cell viability (P < 0.01). DBP exposure for 48 and 96 h significantly lowered the protein expressions of ERβ, BDNF and NPY, and a 96-h exposure significantly reduced the release of the neurotransmitter GABA in the neurons (P < 0.05). DBP exposure causes morphological and functional damages of primary cultured rat hippocampal neurons. DBP-induced neurotoxicity is probably associated with GABA-mediated blockage of the ERβ-BDNF-NPY signaling communication.

Di-butyl phthalate (DBP) induces craniofacial defects during embryonic development in zebrafish.

Di-butyl phthalate (DBP) is commonly added to make plastics softer and more pliable and is found in a variety of consumer and industrial products. Alarmingly high levels of DBP have been detected in water and sediment as DBP leaches from products. These levels are concerning and have led the Environmental Protection Agency to label DBP as a priority environmental pollutant and the European Commission to label DBP as a priority substance. Given the ubiquitous presence of DBP globally and continuous exposure to DBP, studies on the developmental toxicity of DBP are needed. The endocrine disrupting effects of DBP are well documented, but developmental toxicity of DBP during critical developmental time windows is understudied. Here, we investigate the developmental effects of DBP exposure during early development. We find defects in craniofacial development including a decrease in overall cranial size in DBP treated embryos, but the intraocular distance was increased compared to controls. Further investigation of jawbone development demonstrated loss of and disorganization of cartilage development. Defects in vascular innervation and neuronal patterning were also noted. Here we conclude that exposure to DBP during crucial time windows of embryonic development is toxic to craniofacial development.

Environmentally relevant exposure to dibutyl phthalate disrupts DNA damage repair gene expression in the mouse ovary†.

Phthalates have a history of reproductive toxicity in animal models and associations with adverse reproductive outcomes in women. Human exposure to dibutyl phthalate (DBP) occurs via consumer products (7-10μg/kg/day) and medications (1-233μg/kg/day). Most DBP toxicity studies have focused on high supraphysiological exposure levels; thus, very little is known about exposures occurring at environmentally relevant levels. CD-1 female mice (80days old) were treated with tocopherol-stripped corn oil (vehicle control) or DBP dissolved in oil at environmentally relevant (10 and 100μg/kg/day) or higher (1000μg/kg/day) levels for 30days to evaluate effects on DNA damage response (DDR) pathway genes and folliculogenesis. DBP exposure caused dose-dependent effects on folliculogenesis and gene expression. Specifically, animals exposed to the high dose of DBP had more atretic follicles in their ovaries, while in those treated with environmentally relevant doses, follicle numbers were no different from vehicle-treated controls. DBP exposure significantly reduced the expression of DDR genes including those involved in homologous recombination (Atm, Brca1, Mre11a, Rad50), mismatch repair (Msh3, Msh6), and nucleotide excision repair (Xpc, Pcna) in a dose-specific manner. Interestingly, staining for the DNA damage marker, γH2AX, was similar between treatments. DBP exposure did not result in differential DNA methylation in the Brca1 promoter but significantly reduced transcript levels for the maintenance DNA methyltransferase, Dnmt1, in the ovary. Collectively, these findings show that oral exposure to environmentally relevant levels of DBP for 30days does not significantly impact folliculogenesis in adult mice but leads to aberrant ovarian expression of DDR genes.