Sub‑chronic dietary exposure to environmentally relevant doses of di‑(2‑ethylhexyl) adipate (DEHA) induces hepatic steatosis in female mice via disrupting the PPARγ pathway and inducing endoplasmic reticulum stress: In vivo and in vitro evidence.
Aug 2026· Ecotoxicology and Environmental Safety· Vol 323, pp.
120689
· 0 citations· 54 references
Medicine
TL;DR
It is demonstrated that environmental-level DEHA exposure promoted obesity and hepatic steatosis in female mice by disturbing the PPARγ signaling pathway and upregulating ER stress-related genes, highlighting a significant and previously underappreciated health risk of DEHA in females and calling for a reassessment of its safety as an endocrine-disrupting chemical.
Abstract
Di-(2-ethylhexyl) adipate (DEHA), initially regarded as a safer alternative to Di-(2-ethylhexyl) phthalate (DEHP) phthalate plasticizer, is widely used in food packaging and consumer products. Despite its high detection levels in food and established human exposure, the health impacts of DEHA at environmentally relevant doses remain poorly understood. The main aim of this study was to investigate whether sub-chronic dietary exposure to DEHA disrupts metabolism homeostasis in female mice, and to clarify the underlying mechanism through combined in vivo and in vitro models. In this study, 27 four‑week‑old female C57BL/6 J mice with similar initial body weight were randomly assigned to four groups (control (n = 6), vehicle (n = 6), low‑dose DEHA (n = 7), and high‑dose DEHA (n = 8)), where the DEHA concentrations in the mice's diet were 0.087 and 18.2 mg/kg, respectively, for 15 weeks of sub-chronic exposure. DEHA significantly increased body weight, fat mass, fasting blood glucose, serum cholesterol and induced hepatic steatosis, without affecting food intake or energy expenditure. Untargeted metabolomics analysis identified a significant elevation of lipid metabolites in the liver tissues of DEHA-exposed mice. Further mechanistic studies demonstrated that DEHA upregulated the peroxisome proliferator-activated receptor gamma (PPARγ) signaling pathways and the expression of genes associated with endoplasmic reticulum (ER) stress. In both mouse alpha mouse liver 12 (AML12) and human liver-7702 (L02) hepatocytes, DEHA directly induced dose-dependent lipid accumulation. Pharmacological inhibition of ER stress with specific inhibitor 4-phenylbutyric acid (4-PBA) attenuated DEHA-induced lipid accumulation and lipogenic gene expression. These findings demonstrate that environmental-level DEHA exposure promoted obesity and hepatic steatosis in female mice by disturbing the PPARγ signaling pathway and upregulating ER stress-related genes, highlighting a significant and previously underappreciated health risk of DEHA in females and calling for a reassessment of its safety as an endocrine-disrupting chemical.
Phthalates are a class of synthetic compounds, known as endocrine-disrupting chemicals, widely used as plasticizers in consumer products, including personal care items, medical devices, and food packaging. Two common phthalates, di(2-ethylhexyl) phthalate (DEHP) and diisononyl phthalate (DiNP), have been associated with adverse effects on female reproductive health. This study investigated the effects of acute DEHP and DiNP exposure on uterine inflammation and oxidative stress in adult female CD-1 mice. Mice were orally dosed for 10 days with vehicle control, DEHP (20μg/kg/day, 200μg/kg/day, or 200mg/kg/day), or DiNP (20μg/kg/day, 100μg/kg/day, or 200mg/kg/day). Uteri were collected during diestrus for histological and gene expression analyses. Quantitative PCR (qPCR) showed that DEHP (20 and 200μg/kg/day) and DiNP (200mg/kg/day) increased expression of inflammasome-related genes (Il18, Il1β, and Nlrp3). DiNP at 200mg/kg/day also increased Il10 expression. Oxidative stress genes revealed DEHP increased Prdx2 expression at all doses without affecting Sod1, Cat, or Gpx1. However, DiNP increased Prdx2 at 20μg/kg/day but reduced Sod1, Cat, and Gpx1 at higher doses. Histological analysis revealed that high-dose DiNP reduced outer myometrium thickness and luminal epithelial cell height, while DEHP only affected the cell height at the highest dose. Macrophage and other mononuclear phagocytic cell infiltration increased with DEHP (20 and 200μg/kg/day) and all doses of DiNP. while cell proliferation was only changed in DEHP (200μg/kg/day). Together, these findings demonstrate that acute exposure to DEHP and DiNP induces uterine inflammatory and oxidative stress responses, with distinct dose-dependent effects for each phthalate.
Adriana Andrus, L. Y. Parra-Forero, Coba N Sexton et al.· Reproductive Toxicology· 1 citation
Di(2-ethylhexyl) phthalate (DEHP) is widely acknowledged as a prevalent environmental pollutant, mainly due to its extensive use in plastics. This widespread application has resulted in its presence in air, soil, water, and various food items, as well as in agricultural products and tissues of both humans and animals. With the extensive use of plastic products in daily life, the teenage group is facing an increasing risk of DEHP exposure. However, the mechanism of DEHP-induced liver injury, especially during adolescence, remain unclear. This study utilized metabolomics analysis and molecular biology approaches to explore the effects of DEHP on liver function in adolescent mice. We found that DEHP exposure caused hepatic dysfunction, histopathological abnormalities, and lipid deposition. Metabolomics analysis identified the dysregulation of lipid metabolism, especially the fatty acid pathway. Mechanistically, DEHP and its main metabolite, mono(2-ethylhexyl) phthalate (MEHP) drived the ectopic formation of mitochondrial-associated endoplasmic reticulum membranes (MAMs) by up-regulating glucose-regulated protein 75 (GRP75). Such a process is characterized by a reduced interorganellar contact distance between the endoplasmic reticulum and mitochondria as well as elevated expression of MAM-resident proteins. Notably, overexpression of GRP75 alone recapitulated the lipid accumulation phenotype. Conversely, knockdown of GRP75 attenuated MEHP-induced lipid accumulation in AML12 cells. Moreover, a combination of molecular docking, cellular thermal shift assay (CETSA), and drug affinity responsive target stability (DARTS) assays collectively corroborated the direct physical interaction between MEHP and GRP75. Collectively, our findings demonstrate that DEHP/MEHP disrupts the lipid metabolism regulated by MAM through a GRP75-dependent mechanism, thereby leading to liver steatosis. This study clarified the toxicological mechanisms underlying DEHP-induced hepatic dysfunction in adolescent mice, providing new insights into its health risk assessment and potential intervention strategies.
Unknown authors· Food Science and Human Welln...· 0 citations
Environmental factors such as diet and Di(2-ethylhexyl) phthalate (DEHP) have individually been shown to affect lipid metabolism. However, whether DEHP exposure further aggravates metabolic disturbances under high-fat (HF) diet conditions remains insufficiently understood. In this study, we investigated oxidative stress-related responses, lipogenesis- and adipogenesis-related alterations, and liver-adipose tissue changes in rats exposed to DEHP in this model. Metabolic alterations were evaluated by analyzing serum lipid levels, liver and adipose tissue morphology, mRNA expression, enzyme activities, antioxidative defense, and oxidative damage. DEHP-treated HF diet-fed rats showed histopathological alterations in rat liver and adipose tissues. Representative quantitative changes included a 2.64-fold increase in serum TG levels in the DEHP-0.5 group, a 16.64% increase in the liver/body weight ratio in the DEHP-100 group, and a 4.36-fold increase in hepatic MDA levels in the DEHP-100 group. In addition, DEHP exposure altered oxidative stress-related markers and the mRNA expression of lipid metabolism-related genes, such as LDLR. These findings suggest that liver-adipose tissue crosstalk may be involved in metabolic disturbances associated with DEHP exposure in HF diet-fed rats. Further studies including DEHP-only groups under regular diet conditions are needed to clarify the independent effects of DEHP and its potential interactions with dietary fat.
Zhenhua Yang, Jiamin Zhou, Xianyun Zheng et al.· Journal of Environmental Sci...· 0 citations
Phthalates are associated with several reproductive disorders in women and reduce fertility in mice. They are also known to impair hepatic glycogen metabolism. Glucose is a crucial nutrient for the uterus, and glycogen buffers glucose concentration in the endometrium. The objective of this study was to investigate how long-term exposure to di(2-ethylhexyl) phthalate (DEHP) and diisononyl phthalate (DiNP) alters glycogen metabolism in the murine endometrium. Six-week-old female mice were fed chow containing vehicle or DEHP or DiNP at 0.15, 1.5, and 1500 parts per million (ppm) ad libitum for 9 months. Uteri were collected at diestrus. DEHP significantly reduced glycogen levels in the glandular epithelium (GE) and luminal epithelium (LE). In the stroma, both 1.5 and 1500 ppm groups had significantly lower glycogen. In the DiNP-treated mice, all three concentrations significantly decreased glycogen in GE, LE, and stroma. Neither phthalate altered mRNA levels of hexokinase1 (Hk1), glycogen synthase 1 (Gys1), glycogen phosphorylase M (Pygm), or glucose-6-phosphatase 3 (G6pc3). Immunohistochemistry showed that both phthalates increased HK1 levels in the stroma but not the epithelium. DEHP and DiNP (1500 ppm) increased PYGM in GE, LE, and stroma. DiNP (1500 ppm) significantly lowered G6PC3 in LE compared to all other groups. In the GE, both 1.5 and 1500 ppm DiNP decreased the immunostaining of G6PC3 compared to control and 0.15 ppm DiNP. Our results show that phthalates alter endometrial glycogen levels and expression of key enzymes. These findings are consistent with altered glycogen metabolism, which could alter endometrial glucose metabolism.
The plasticizer butyl benzyl phthalate (BBP), a ubiquitous endocrine disruptor, is commonly used in PVC products and can leach into the environment, leading to human exposure. BBP has been detected in the placenta, amniotic fluid, and breast milk, indicating potential exposure during gestation and lactation. BBP is an endocrine disruptor compound (EDC) with estrogenic and anti-androgenic activity, which, at high or acute doses, may interfere with the development of hormone-dependent systems, including components of the neuro-immune-endocrine network (NIE) and sexual dimorphism. This study aimed to investigate the effects of perinatal BBP exposure on various components of the NIE network in adult male and female rats. Pregnant rats were administered drinking water with or without BBP (50 μg/L) from gestational day 5 until weaning. Offspring were assessed at 9 weeks of age by examining sex hormone levels in serum, immune cell populations in the spleen, neurotransmitters in the hippocampus, and cytokines in serum, hippocampus, and spleen. Results showed that BBP exposure led to lower weaning weights in both sexes, with persistent effects in males. Splenic T and T-helper lymphocyte populations increased in both sexes. In the hippocampus, females exhibited elevated proinflammatory cytokines, while males showed decreased serotonin levels, indicating sex-specific alterations in hormone, cytokine, and neurotransmitter levels in adult offspring.
D. L. Ruiz-Antonio, K. Nava-Castro, C. Garay-Canales et al.· Frontiers in Immunology· 0 citations
