Synergistic assault of DEHP and MPs: Unmasking the ER stress-triggered autophagic injury male fertility.

Combined toxicity influence of polypropylene microplastics and di-2-ethylhexyl phthalate on physiological-biochemical characteristics of cucumber (Cucumis sativus L.)

Microplastics and di (2-ethylhexyl) phthalate synergistically induce apoptosis in mouse pancreas through the GRP78/CHOP/Bcl-2 pathway activated by oxidative stress

Combined Enterohepatic Toxicity of Polystyrene Microplastics and Di(2-ethylhexyl) Phthalate in Mice: Gut Microbiota-Dependent Synergistic Effects.

Di-2-ethylhexyl phthalate (DEHP) exposure induces sperm quality and functional defects in mice

Rationale: Accumulated evidence indicates that environmental plasticizers are a threat to human and animal fertility. Di (2-ethylhexyl) phthalate (DEHP), a plasticizer to which humans are exposed daily, can trigger reproductive toxicity by acting as an endocrine-disrupting chemical. In mammals, the female primordial follicle pool forms the lifetime available ovarian reserve, which does not undergo regeneration once it is established during the fetal and neonatal period. It is therefore critical to examine the toxicity of DEHP regarding the establishment of the ovarian reserve as it has not been well investigated.Methods: The ovarian cells of postnatal pups, following maternal DEHP exposure, were prepared for single cell-RNA sequencing, and the effects of DEHP on primordial follicle formation were revealed using gene differential expression analysis and single-cell developmental trajectory. In addition, further biochemical experiments, including immunohistochemical staining, apoptosis detection, and Western blotting, were performed to verify the dataset results.Results: Using single-cell RNA sequencing, we revealed the gene expression dynamics of female germ cells and granulosa cells following exposure to DEHP in mice. Regarding germ cells: DEHP impeded the progression of follicle assembly and interfered with their developmental status, while key genes such as Lhx8, Figla, and others, strongly evidenced the reduction. As for granulosa cells: DEHP likely inhibited their proliferative activity, and activated the regulation of cell death. Furthermore, the interaction between ovarian cells mediated by transforming growth factor-beta signaling, was disrupted by DEHP exposure, since the expression of GDF9, BMPR1A, and SMAD3 was affected. In addition, DNA damage and apoptosis were elevated in germ cells and/or somatic cells.Conclusion: These findings offer substantial novel insights into the reproductive toxicity of DEHP exposure during murine germ cell cyst breakdown and primordial follicle formation. These results may enhance the understanding of DEHP exposure on reproductive health.

Background: Di(2-ethylhexyl) phthalate (DEHP) is the most widely used polyvinyl chloride (PVC) plasticizer. Therefore, DEHP pollution is spreading all over the world. In recent years, it has attracted attention as an endocrine disrupting chemical. In animal experiments using rodents, testicular toxicity has been confirmed by feeding diets containing DEHP. Mono(2-ethylhexyl) phthalate (MEHP), a potent oxidative stressor, is thought to be directly involved in testicular toxicity. On the other hand, caffeine and ethanol, which are hydroxyl radical scavengers, are taken in relatively large amounts from food and drink on a daily basis. Although the reproductive toxicity of DEHP in humans remains unclear, it is important to clarify the relationship between DEHP toxicity and caffeine and ethanol intake through animal experiments. Method: To investigate the effects of caffeine and ethanol intake on DEHP-induced testicular atrophy, rats were fed a 1 w/w % DEHP diet and caffeinated water (0.05 w/w %) or ethanol water (2.5 or 5 v / v %) or ethanol plus caffeine water for one week. Result: By administering 1 w/w % DEHP-containing diet for one week, significant testicular weight decrease noted in all cases, and testicular MEHP concentration in the control group and DEHP–only administered group showed a strong negative correlation with the relative testicular weight (as a percentage of body weight). This indicates that there is a close relationship between the tissue MEHP concentration and testicular atrophy in the target organ, the testis. In addition, it was found that the increase in Fe, Cu, and Ca concentrations in the testis was closely related to the decrease in relative testicular weight. Furthermore, multiple linear regression analysis suggested that the MEHP concentration in the testis was significantly related to the increase in the Cu concentration in the testis. This suggests that MEHP oxidative stress may induce the SOD enzyme. Also, caffeine administration was found to slightly but significantly improve Zn levels. On the other hand, caffeine treatment significantly inhibited the decrease in testicular weight. Also, the testicular weight decrease in the ethanol and caffeine administration groups was less than that in the DEHP-only administered group. These findings suggest that ethanol and caffeine may inhibit DEHP testicular atrophy. In addition, in this administration experiment, the relative testicular weight vs. testicular MEHP plot in the ethanol and caffeine administered groups was clearly distributed above the regression line obtained from the relationship between the relative testicular weight and testicular MEHP concentration in the control and DEHP-only administered groups. From this, it was suggested that administration of DEHP in combination with ethanol or caffeine reduced DEHP testicular toxicity. Furthermore, multiple linear regression analysis suggested that combined use of ethanol and caffeine had a combined effect of reducing testicular atrophy. Conclusion: Daily use levels of caffeine and ethanol were found to reduce DEHP-induced testicular toxicity in rats

Di-(2-ethyl hexyl) phthalate induced oxidative stress promotes microplastics mediated apoptosis and necroptosis in mice skeletal muscle by inhibiting PI3K/AKT/mTOR pathway

Exposure to Phthalate Esters

Several members of the phthalate ester family have antiandrogenic properties, yet little is known about how exposure to these ubiquitous environmental contaminants early in development may affect sexual development. We conducted experiments to determine effects of in utero and lactational exposure to the most prevalent phthalate ester, di(2-ethylhexyl) phthalate (DEHP), on male reproductive system development and sexual behavior. Sprague-Dawley rats were dosed with corn oil or DEHP (0, 375, 750, or 1,500 mg/kg/day, per os) from gestation day 3 through postnatal day (PND) 21. Dose-related effects on male offspring included reduced anogenital distance, areola and nipple retention, undescended testes, and permanently incomplete preputial separation. Testis, epididymis, glans penis, ventral prostate, dorsolateral prostate, anterior prostate, and seminal vesicle weights were reduced at PND 21, 63, and/or 105-112. Additional dose-related effects included a high incidence of anterior prostate agenesis, a lower incidence of partial or complete ventral prostate agenesis, occasional dorsolateral prostate and seminal vesicle agenesis, reduced sperm counts, and testicular, epididymal, and penile malformations. Many DEHP-exposed males were sexually inactive in the presence of receptive control females, but sexual inactivity did not correlate with abnormal male reproductive organs. These results suggest that in utero and lactational DEHP exposure also inhibited sexually dimorphic central nervous system development. No major abnormalities were found in any of eight control litters, but DEHP caused severe male reproductive system toxicity in five of eight litters at 375 mg/kg/day, seven of eight litters at 750 mg/kg/day, and five of five litters at 1,500 mg/kg/day. These results demonstrate that the male reproductive system is far more sensitive to DEHP early in development than when animals are exposed as juveniles or adults. The effects of DEHP on male reproductive organs and sexual behaviors and the lack of significant effects on time to vaginal opening and first estrus in their littermates demonstrate that DEHP (and/or its metabolites) affects development of the male reproductive system primarily by acting as an antiandrogen. The pattern of effects of in utero and lactational DEHP exposure differed from patterns caused by other phthalate esters, and the preponderance of anterior prostate agenesis appears to be unique among all chemicals. These results suggest that DEHP acts partly by mechanisms distinct from those of other antiandrogens.

Several members of the phthalate ester family have antiandrogenic properties, yet little is known about how exposure to these ubiquitous environmental contaminants early in development may affect sexual development. We conducted experiments to determine effects of in utero and lactational exposure to the most prevalent phthalate ester, di(2-ethylhexyl) phthalate (DEHP), on male reproductive system development and sexual behavior. Sprague-Dawley rats were dosed with corn oil or DEHP (0, 375, 750, or 1,500 mg/kg/day, per os) from gestation day 3 through postnatal day (PND) 21. Dose-related effects on male offspring included reduced anogenital distance, areola and nipple retention, undescended testes, and permanently incomplete preputial separation. Testis, epididymis, glans penis, ventral prostate, dorsolateral prostate, anterior prostate, and seminal vesicle weights were reduced at PND 21, 63, and/or 105-112. Additional dose-related effects included a high incidence of anterior prostate agenesis, a lower incidence of partial or complete ventral prostate agenesis, occasional dorsolateral prostate and seminal vesicle agenesis, reduced sperm counts, and testicular, epididymal, and penile malformations. Many DEHP-exposed males were sexually inactive in the presence of receptive control females, but sexual inactivity did not correlate with abnormal male reproductive organs. These results suggest that in utero and lactational DEHP exposure also inhibited sexually dimorphic central nervous system development. No major abnormalities were found in any of eight control litters, but DEHP caused severe male reproductive system toxicity in five of eight litters at 375 mg/kg/day, seven of eight litters at 750 mg/kg/day, and five of five litters at 1,500 mg/kg/day. These results demonstrate that the male reproductive system is far more sensitive to DEHP early in development than when animals are exposed as juveniles or adults. The effects of DEHP on male reproductive organs and sexual behaviors and the lack of significant effects on time to vaginal opening and first estrus in their littermates demonstrate that DEHP (and/or its metabolites) affects development of the male reproductive system primarily by acting as an antiandrogen. The pattern of effects of in utero and lactational DEHP exposure differed from patterns caused by other phthalate esters, and the preponderance of anterior prostate agenesis appears to be unique among all chemicals. These results suggest that DEHP acts partly by mechanisms distinct from those of other antiandrogens.

Microplastics (MPs) and the plasticizer di(2-ethylhexyl) phthalate (DEHP) frequently co-occur, presenting substantial health risks to both humans and animals. While animal studies indicate adverse effects from exposure to MPs and DEHP, their potential toxicity in humans remains uncertain. This study examines the response of human hepatocellular carcinoma (HepG2) cells to concurrent exposure to synthetic spherical polystyrene (PS) particles and DEHP. We analyzed the effect of particle size on the internalization of PS-MPs using HepG2 spheres as a 3D model. The results showed that MPs at 100 nm had the highest internalization efficiency, which gradually decreased as the particle size increased to 1 and 5 μm. In addition, DEHP significantly improved the internalization of MPs, especially for 5 μm particles, which showed a 26% increase in internalization efficiency. We also evaluated changes in physiological activity. Co-exposure to MPs and DEHP resulted in significantly higher cytotoxicity than exposure to MPs alone, with a 20% reduction in cell viability. Larger particle sizes led to greater cellular damage, indicated by a 20% increase in reactive oxygen species (ROS) and a 40% rise in lactate dehydrogenase (LDH) release, suggesting membrane rupture. This study offers new insights into the potential toxicity of short-term exposure to MPs and DEHP, using HepG2 spheres to closely replicate invivo conditions.

Combined negative effects of microplastics and plasticizer DEHP: The increased release of Nets delays wound healing in mice

Di(2-ethylhexyl) phthalate (DEHP) is ubiquitous environmental contaminant and identified as endocrine-disrupting chemical (EDC), present in plastics as plasticizer. Due to its versatile use, human exposure level reaches to danger limit. The main focus of our study is to see the effect of vitamin C on hematological and biochemical alterations caused by Di(2-ethylhexyl) Phthalate toxicity in female albino mice, Mus musculus. It is found to cause defects of the liver, kidney, and lungs. Its anti-androgenic nature brings the main focus on its toxicity associated with reproductive and endocrine system. In this experimental study, 18 young female Swiss albino mice, Mus musculus, were used and divided into 3 groups of 6 animals each as control (corn oil vehicle), DEHP group (100 mg/kg body weight dissolved in corn oil), and DEHP + vitamin-C group (100 mg/kg body weight each, dissolved in corn oil and double distilled water, respectively) for 90 days. In this research, serum metabolites were evaluated to study the effect of DEHP on glucose, total protein, and lipid profile along with some hematological, enzymological, and oxidative stress parameters. Simultaneously, we compared the effectiveness of vitamin-C against DEHP toxicity to mitigate the serum homeostasis disturbance. In present study, we observed, in DEHP-treated animals, glucose, triglycerides, very-low-density lipoprotein (VLDL), total protein, alkaline phosphatase (ALP), acid phosphatase (ACP), and alanine aminotransferase (ALT) levels increased remarkably, whereas total cholesterol, high-density lipoproteins (HDL), aspartate aminotransferase (AST), total RBC count, total WBC count, and hemoglobin (Hb) level significantly decreased as compared to control group. In addition, we noticed there was a decrease in superoxide dismutase (SOD) and increase in levels of lipid peroxidation (MDA) and interleukin-6 (IL-6) in DEHP treatment group as compared to control group. The results indicated vitamin C had a better improving effect against DEHP toxicity on balancing metabolic abnormalities and inflammation-related comorbidities.

Objective To study reproductive functional lesions of Di (2-ethylhexyl) Phthalate(DEHP) on epididymis, and to find antagon which can repress the toxicity. Methods In vivo study,postnatal day 20 (PND20) and PND50 male KM mice were randomly divided into normal control, cornoil, DEHP and DEHP + Zinc gluconate group. In each stage and group, after been garaged for 10days, epididymal histopathologic changes and androgen receptor (A.R) and estrogen receptor (ER) inepididymis were detected. In vitro study, epididymal epithelial cells of PND20 and PNDS0 mice werecultured. Viability of epididymal epithelial ceils were measured using MTT assay, content of sialic acid(SA) and the activity of α-1,4-glucosidase and lactic acid dehydrogenase (LDH) were investigated. Re-sults DEHP induced epididymis atrophy and conspicuous histopathologic uhrastructure changes, up-regulated AR and down-regulated ER. No conspicuous variations were found in DEHP + Zinc gluco-hate group. Conclusions DEHP can induce lesions of epididymaI structure and function. Zinc is an an-tagon to DEHP toxicity. Key words: Dicthylhexyl phthalate; Reproduction; Epididymis; Zinc; Mice

Combined toxicity of polystyrene microplastics and ammonium perfluorooctanoate to Daphnia magna: Mediation of intestinal blockage