N-acetylcysteine and zinc sulphate abate di-2-ethylhexyl phthalate-mediated reproductive dysfunction in rats:Focus on oxidative and sex hormone receptors mechanisms

Objective:To investigate the potential of N-acetylcysteine(NAC)and zinc sulphate(ZnSO_(4))in mitigating reproductive dysfunction caused by di-2-ethylhexyl phthalate(DEHP)in rats and to understand the underlying mechanisms,specifically oxidative stress and sex hormone receptor activity.Methods:Thirty-five male Wistar rats were randomly divided into five equal groups(n=7 per group).Group 1 was administered 0.5 mL of distilled water and served as the control group.Group 2 was given only DEHP(750 mg/kg/day),while group 3,4 and 5 were given DEHP(750 mg/kg/day)plus NAC(100 mg/kg/day),DEHP(750 mg/kg/day)plus ZnSO_(4)(0.5 mg/kg/day),and DEHP(750 mg/kg/day)plus NAC(100 mg/kg/day)as well as ZnSO_(4)(0.5 mg/kg/day),respectively.All treatments lasted for 21 days.Samples were obtained after the rats were sacrificed,and hormones levels in the serum and markers of oxidative stress in the testicles were analyzed using the enzyme-linked immunosorbent assay.The amount of androgen receptors in the testicles was determined by immunohistochemistry,and the susceptibility of testosterone and DEHP to bind to androgen receptor and 5α-reductase was determined by molecular docking studies.Results:DEHP decreased reproductive hormones,testicular antioxidant enzymes,increased malondialdehyde levels,and negatively impacted histology of the pituitary and testes.NAC or ZnSO_(4) treatment showed a marked improvement in testicular antioxidant status and hormone levels,as well as a positive effect on the histology of the pituitary and testes.The combination of both treatments appeared to be more effective.The affinity of DEHP to bind to androgen receptors may lead to disruption of androgen receptor signaling,which can further result in dysfunction of hormones related to androgen.However,NAC is more likely to form stronger binding interactions with follicle stimulating hormone and luteinizing hormone receptors,as well as gonadotropin-releasing hormone receptors,when compared to DEHP.Conclusions:The possibility that NAC and ZnSO_(4) could downregulate DEHP-induced sex hormone changes is suggested by their potential to reduce toxicity.

Urine Metabonomic Analysis of Interventions Effect of Soy Isoflavones on Rats Exposed to Di-(2-ethylhexyl) Phthalate

Objective Di-(2-ethylhexyl) phthalate(DEHP) is a ubiquitous environmental contaminant.As an endocrine disruptor,it seriously threatens human health and ecological environmental safety.This study examines the impact of intervention with soybean isoflavones(SIF) on DEHP-induced toxicity using a metabonomics approach.Methods Rats were randomly divided into control(H),SIF-treated(A,86 mg/kg body weight),DEHP-treated(B,68 mg/kg),and SIF plus DEHP-treated(D) groups.Rats were given SIF and DEHP daily through diet and gavage,respectively.After 30 d of treatment,rat urine was tested using UPLC/MS with multivariate analysis.Metabolic changes were also evaluated using biochemical assays.Results Metabolomics analyses revealed that p-cresol glucuronide,methyl hippuric acid,N1-methyl-2-pyridone-5-carboxamide,lysophosphatidycholine [18:2(9 Z,12 Z)] {lyso PC [18:2(9 Z,12 Z)]},lyso PC(16:0),xanthosine,undecanedioic acid,and N6-acetyl-l-lysine were present at significantly different levels in control and treatment groups.Conclusion SIF supplementation partially protects rats from DEHP-induced metabolic abnormalities by regulating fatty acid metabolism,antioxidant defense system,amino acid metabolism,and is also involved in the protection of mitochondria.

An Estimation of the Daily Intake of Di(2-ethlhexyl) Phthalate(DEHP) among Workers in Flavoring Factories

Objective To estimate the daily intake of DEHP among workers in flavoring factories. Methods 71 workers in two flavoring manufacturers, 27 administrators in those factories and 31 laboratory technicians in a research institute were recruited and assigned to exposure group, control group 1 and control group 2 respectively. Their urinary DEHP metabolites, mono（2-ethylhexyl） phthalate （MEHP）, mono-（2-ethyl-5-hydroxyhexyl） phthalate （MEHHP）, mono-（2-ethyl-5-oxohexyl） phthalate （MEOHP）, were detected by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. The urinary metabolites concentrations were converted into DEHP intake levels using two pharmacokinetic models： the urine creatinine-excretion （UCE） one and the urine volume （UV） one. Results No significant differences were found among the three groups. Based on the urinary concentrations of Z3MEHP, we got a median daily DEHP intake of 3.22 or 1.85 μg/kg body-weight/day applying the UV or UCE models respectively. Depending on the UV model, three subjects （2.34%） exceeded the RfD value given by US EPA and the P50 of estimate daily DEHP intakes accounted for 16.10% of the RfD value. No subjects exceeded the limitation depending on the UCE model. Conclusion The workers in flavoring factories were not supposed to be the high DEHP exposure ones and their exposure level remained at a low risk.

Impairment of Di(2-Ethylhexyl) Phthalate on Cellular Immunity in Kunming Mice

Immunity is crucial to the health of animals and it can determine their survival and fitness. Di(2-ethylhexyl) phthalate (DEHP) is widely used as a plasticizer and hence is the most abundant phthalate in the environment. Exposure to DEHP is of great concern for human health. In the present study, we tested the hypothesis that exposure to DEHP would suppress T cell-mediated immunity in mice. Twenty adult male Kunming mice were randomly assigned into the control (n = 10) and the DEHP treatment (n = 10) groups. Both groups have free access to food and water, while the mice in the latter group drank DEHP solution (2000 mg/L) for 42 days. T cell-mediated immunity assessed by phytohaemagglutinin (PHA) response was depressed in the DEHP treated mice compared with the controls, however, wet thymus and spleen mass, white blood cells were not influenced by DEHP treatment. Taken together, different immunological parameters responded differently to DEHP treatment in Kunming mice.

Association of Di(2-ethylhexyl)Phthalate Exposure with Reproductive Hormones in the General Population and the Susceptible Population:A Systematic Review and Meta-Analysis

Di(2-ethylhexyl)phthalate(DEHP),an environ-mental endocrine disruptor,has hormone-like activity and endocrine-disrupting effects.However,the types of reproductive hormones associated with DEHP vary across the studies.Thus,we conducted a systematic review and meta-analysis to pool existing epidemiological evidence.We searched three databases up to January 31,2024,for eligible original studies to ultimately include 37 studies from eight countries with a total of 28911 participants.DEHP exposure was evaluated with urinary metabolites.Since the main types,production sites,blood concentrations,and functions of reproductive hormones differ between men and women,we reported the combined effect values by gender.Subgroup analyses were conducted by age,subfertility status,and the national sociodemographic index(SDI)level.Furthermore,the effect of maternal exposure during pregnancy on children’s reproductive hormone levels was analyzed separately.Overall,in general,in men,DEHP was positively correlated with sex hormone binding-globulin(SHBG)and adversely correlated with total testosterone(TT),free androgen index(FAI),and follicle-stimulating hormone(FSH).Results indicated that among men of reproductive age,DEHP exposure was associated with more significant hormonal suppression in infertile men compared with fertile men.Notably,age subgroup analysis among women revealed that postmenopausal women were more vulnerable to DEHP,which was related to lower TT and estradiol(E2).However,this study did not observe a significant association between prenatal DEHP metabolites and reproductive hormone levels in children.Our research identifies the most susceptible hormones(androgen suppression)after DEHP exposure and suggests that infertile men and postmenopausal women are in great need of more attention as sensitive populations.

Aquatic toxicity of di (2-eihylhexyl) phthalate to duckweeds

This study is concerned with the effects of di （2-ethylhexyl） phthalate （DEHP） on two kinds of duckweeds （Spirodela polyrhiza and Lemna minor）.The results indicate that DEHP has aquatic toxicity to Spirodela polyrhiza at 0.4 mg/L and to Lemna minor at over 0.1 mg/L by changing their physiologic-biochemical characteristics.The contents of duckweed chlorophyll and soluble protein decrease with increasing DEHP concentration after 7 d of exposure.DEHP shows the stimulating role in catalase （CAT） and superoxide dismutase （SOD） systems at relative low levels.At 0.01 mg/L and 0.005 mg/L,SOD activities of Spirodela polyrhiza and Lemna minor reach their peak values respectively,while CAT activity reaches its maximum value at 0.05 mg/L and 0.01 mg/L.When DEHP levels are too high,the protection enzyme system would be destroyed and plant growth is inhibited.The analysis of malondialdehyde （MDA） and Fourier transform infrared spectroscopy manifest that DEHP could affect the tested duckweeds by destroying its cell membranes,and Spirodela polyrhiza is more resistant to DEHP exposure than Lemna minor.

Primary neuronal-astrocytic co-culture platform for neurotoxicity assessment of di-(2-ethylhexyl) phthalate

Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding environment. This study focused on DEHP＇s effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species （ROS） concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation （gliosis） was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uotake and effects on the human neuronal system from exoosure to DEHP in the indoor environment.

Interaction Between DEHP and Particulate in a Eutrophic Lake

Characteristics of interaction between di-2-ethylhexyl phthalate(DEHP) and particulate in a eutrophic lake were studied in this paper. DEHP concentrations ranged from 89.9 to 247 μg/L with an average value of 146 μg/L in subsurface water (SSW) samples, and from 82.0 to 390 μg/L with an average value of 211 μg/L in water surface microlayer (SM) samples. The results indicate that there was only a weak correlation between the DEHP concentrations and suspended particulate material(SPM) concentrations in both SSW and SM, while the significant correlation between DEHP concentrations and chlorophyll a concentrations was found, which suggestes that DEHP was principally bound to phytoplankton in the eutrophic lake. Correlation between DEHP concentrations and total phosphor (TP) concentrations was also found in this investigation.Enrichment factors (EF) of DEHP in SM were in the range of 0.85 to 2.12 with an average value of 1.35. DEHP EFs were significantly related to the enrichment of chlorophyll a in SM. The results suggest that the enrichment of DEHP in SM of this eutrophic lake was mainly due to DEHP accumulation in phytoplankton and was controlled by distribution of phytoplankton between SM and SSW.

Plastic Additives Decrease Agrin-Induced Acetylcholine Receptor Clusters and Myotube Formation in C2C12 Skeletal Muscle Cell Culture

Common additives in plastics such as bisphenol A (BPA) or phthalates like di-(2-ethylhexyl) phthalate (DEHP) are environmental estrogens that have been shown to be endocrine disruptors in some experimental animal models. This project used the C2C12 cell culture model to examine how exposure to BPA or DEHP affects two aspects of skeletal muscle development, the fusion of myoblasts into myotubes and agrin-induced clustering of acetylcholine receptors (AChRs). During myotube formation AChRs cluster spontaneously. Treatment with motor neuron derived agrin increases the frequency of AChR clusters through an agrin signaling pathway that also clusters other postsynaptic components of the neuromuscular synapse. For this project C2C12 cells were exposed to BPA or DEHP while myoblasts fused into myotubes. After exposure to 10 μM BPA or 100 μM DEHP the frequency of agrin-induced AChR clusters decreased. In addition, myotube formation decreased as a higher percentage of nuclei remained in myoblasts. Furthermore, BPA or DEHP reduced the amount of the myogenic regulatory factor myogenin. This suggests that BPA and DEHP decrease AChR clustering by reducing myogenin. Moreover, plastic additives like BPA and DEHP may pose a risk for skeletal muscle development in humans.

Unveiling the Nexus the link between water quality index and phthalic acid ester concentrations in Tigris River

Monitoring water quality in an aquatic system is a crucial objective in pursuing sustainable development,despite the challenges posed by water scarcity and the impact of climate change.The objective of the present study was to examine the water quality of the Tigris River along its course from the point of entry into Baghdad city to its exit,particularly in the presence of Phthalic Acid Esters(PAEs).In both dry and wet seasons and from five sites of the river,nine environmental factors(Turbidity,Total dissolved solids,pH,dissolved oxygen,Chloride,nitrate,phosphate,sulfate,and Biological oxygen demand)and three PAEs(di-n-butyl phthalate(DBP),di-isononyl phthalate(DiNP),and di-2-ethyl hexyl phthalate(DEHP))were investigated.Results revealed that the river has high levels of turbidity and Total dissolved solids,slightly alkaline,and well-aerated(dissolved oxygen>7 mg/L).Chloride,nitrate,phosphate,and sulfate values varied between seasons,as high values were recorded in the wet season.Biochemical oxygen demand levels were higher during the dry season.The highest value of total PAEs(2.20μg/L)was observed during the dry season,and the lowest value was in the wet season(1.16μg/L).The HQ results revealed that the river was ranked as a moderate risk area.However the water quality index and environmental risk index classified the water quality as a poor category,with a high-risk score in the study sites within Baghdad city.Therefore,despite the acceptable limit of PAEs in the river's water(<3μg/L),its presence is an early alarm to withdraw attention to its environmental risk.These findings highlight the need for appropriate measures to control and remove pollutants to ensure a safe and healthy water supply in the Tigris River.