Di (2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer known to cause testicular toxicity, though its mechanism remains incompletely understood. This study investigated whether DEHP-induced testicular injury involved suppression of androgen receptor (AR) expression. In vivo, rats were exposed to DEHP (500 mg/kg) for 60 days, causing damage to the testicles. Further, the expression of AR was decreased, mitochondrial quality control (MQC) was impaired, mitochondrial damage occurred, and ultimately led to ferroptosis. Specifically, DEHP reduced levels of mitochondrial biogenesis markers (PGC-1α, TFAM, NRF1/2) and fusion markers (MFN1/2, OPA1), while increasing fission markers (DRP1, FIS1) and mitophagy markers (PINK1, PARKIN). In vitro, to further explore the relationship between AR, MQC, and ferroptosis, the TM4 cell model overexpressing AR was established and exposed to MEHP (200 μM). The findings demonstrated that AR overexpression effectively alleviated DEHP-induced disruption of MQC, mitochondrial impairment, and ferroptosis. Together, these findings demonstrated that DEHP impaired the MQC system by inhibiting AR expression, thereby promoting ferroptosis.

Di-(2-ethylhexyl) phthalate (DEHP) is a plasticizer widely used to enhance the flexibility and durability of plastic products. As an environmental endocrine disruptor, DEHP impairs male reproductive function. Its metabolite, mono-(2-ethylhexyl) phthalate (MEHP), mediates many toxic effects, but the mechanisms remain unclear. We hypothesized that DEHP induces testicular necroptosis through MEHP-mediated calcium overload and the RIPK1-regulated lysosomal-mitochondrial axis. Here, we reveal this novel mechanism. This study investigated DEHP-induced testicular damage, focusing on necroptosis and calcium (Ca²⁺) signaling pathways. Sprague-Dawley rats were exposed to 250 and 750 mg/kg DEHP for 5 weeks. Testicular damage was assessed via histopathology, testosterone measurement, and RNA sequencing (RNA-seq). A common Sertoli cell line was treated with MEHP to study Ca²⁺ overload, lysosomal membrane permeabilization (LMP), mitochondrial dysfunction, and necroptosis. Pharmacological inhibitors were employed to explore pathway involvement, including CA-074 Me (cathepsin B inhibitor), BAPTA-AM (Ca²⁺ chelator), and Nec-1 (RIPK1 inhibitor). DEHP caused testicular damage, including seminiferous tubule disorganization and reduced plasma testosterone. RNA-seq revealed necroptosis pathway enrichment, with upregulated RIPK1, RIPK3, MLKL, and PGAM5. MEHP induced Ca²⁺ overload, LMP, and mitochondrial dysfunction in Sertoli cells. CA-074 Me attenuated mitochondrial damage, while BAPTA-AM mitigated LMP. Nec-1 suppressed necroptosis-related proteins and restored blood-testis barrier integrity by upregulating ZO-1, Cx-43 and Claudin-11. DEHP exposure induced testicular necroptosis via MEHP-mediated Ca²⁺ overload-lysosomal-mitochondrial axis, regulated by RIPK1. These findings provide insights into DEHP reproductive toxicity.

Arsenic exposure, a typical environmental stressor, is closely associated with nonalcoholic steatohepatitis (NASH), but the definite mechanism remains elusive. The integrated stress response (ISR) acts as a core signaling cascade that mediates cellular stress responses and is implicated in the development of multiple metabolic disorders. Nevertheless, the critical regulatory role of ISR in the progression of arsenic-associated NASH has not been definitively clarified. In the present study, we verified the activation of ISR in arsenic-induced NASH by detecting the expression of ISR-related markers through in vivo and in vitro. Notably, the majority of the downstream impacts of the ISR were modified after arsenic exposure. However, of the four upstream ISR signaling initiators, only Protein Kinase R-like Endoplasmic Reticulum Kinase (PERK) was influenced, as evidenced by a marked elevation in PERK phosphorylation levels following arsenic treatment. Furthermore, we demonstrated that NaAsO2 downregulated the protein levels of multiple coagulation factor deficiency protein 2 (MCFD2), which is localized on the endoplasmic reticulum and Golgi apparatus, in vivo and in vitro. Notably, overexpression of MCFD2 markedly attenuated PERK-eIF2α-mediated ISR, inflammation and lipid accumulation caused by arsenic in vitro. In conclusion, our findings reveal that arsenic exposure triggers the activation of PERK-eIF2α-mediated ISR and NASH by suppressing MCFD2. These findings may provide insights into the underlying mechanisms of NASH.

The persistent organic pollutant perfluorooctane sulfonate (PFOS) has been shown to induce hepatocyte pyroptosis. Although N-terminal domain of Gasdermin D (GSDMD-N) is a well-established executor of pyroptotic pore formation at the plasma membrane (PM), the origin of PM-localized GSDMD-N remains unclear in PFOS-induced pyroptosis. This study showed that PFOS elevated the levels of total GSDMD-N in human hepatocytes HepG2 and mice liver. In the PFOS-treated cells, mitochondrial GSDMD-N rose from 12 h. At 24 h, we noticed that mitochondrial GSDMD-N did not increase further but decreased instead, with a corresponding increase observed in PM GSDMD-N. Mitochondrial GSDMD-N caused membrane perforation and rupture. Subsequently, ATP synthase subunit f (ATP5J2) redistributed from mitochondria to PM. Notably, the translocation of ATP5J2 paralleled that of GSDMD-N. Knockdown of ATP5J2 induced GSDMD-N to accumulate in mitochondria while withdraw from the PM, exacerbating mitochondrial membrane damage. Further investigation confirmed the interaction of ATP5J2 and GSDMD-N in PFOS-exposed HepG2 cells and mice liver. Here, we demonstrate that under PFOS exposure, PM GSDMD-N originates from mitochondrial GSDMD-N, which is mediated by ATP5J2. Our research enables deeper understanding of mitochondria-dependent pyroptosis and provides novel mechanistic insights into PFOS toxicity.

Perfluorooctane sulfonate (PFOS), classified as a persistent organic pollutant, promotes mitochondrial iron overload through voltage-dependent anion channel 2 (VDAC2). However, the source of iron transported through VDAC2 remains unclear. Here, we reported that PFOS enhanced extracellular iron influx into human hepatocytes HepG2, which peaked at 6 h. Under PFOS exposure, total VDAC2 protein levels remained unchanged, whereas its expression increased in mitochondrial lysates but decreased in cytoplasmic lysates, which were observed both in HepG2 cells and mice liver. Meanwhile, the total protein expression of the extracellular iron importer ZRT/IRT-like protein 14 (ZIP14) was upregulated, whereas absent from the mitochondrial lysates. Knockdown of either VDAC2 or ZIP14 inhibited PFOS-induced extracellular iron influx and mitochondrial iron overload, but did not affect cytoplasmic iron levels. Notably, immunofluorescence colocalization analysis revealed that the timeline of mitochondrial redistribution toward the plasma membrane closely paralleled the course of increased extracellular iron influx. Furthermore, the result of molecular docking suggested the potential interaction between VDAC2 and ZIP14. Together, these works pointed out and further advanced our understanding of the extracellular iron transport into mitochondria in hepatocytes under PFOS exposure and revealed a potential therapeutic target for mitigating the disease mediated by iron-related mechanism.

Inorganic arsenic exposure remains a global public health concern, and its toxicological outcomes are highly dependent on biotransformation processes within the body. Among these processes, arsenic methylation—catalyzed predominantly by arsenic methyltransferase (AS3MT)—represents a critical determinant of arsenic disposition, toxicity, and disease susceptibility. AS3MT-mediated methylation converts iAs into mono- and dimethylated arsenicals, metabolites whose biological effects vary markedly depending on exposure dose, duration, and individual genetic background. Experimental studies indicate that under high-dose, short-term exposure, AS3MT-driven methylation facilitates arsenic clearance and exerts a detoxifying effect. In contrast, epidemiological evidence demonstrates that chronic low-dose exposure promotes the accumulation of toxic methylated metabolites, thereby enhancing the risk of metabolic disorders, cardiovascular diseases, neurotoxicity, and multiple cancers. Genetic polymorphisms of AS3MT further modulate methylation efficiency, metabolite profiles, and downstream molecular responses, including epigenetic dysregulation, oxidative stress, and inflammatory signaling, ultimately shaping individual susceptibility to arsenic-induced diseases. Accumulating evidence links specific AS3MT variants to altered risks of adverse outcomes, underscoring its dual role as both a detoxification enzyme and a mediator of arsenic toxicity. This review systematically summarizes current evidence on the mechanistic roles of AS3MT and its genetic polymorphisms in iAs methyl metabolism and arsenic-related disease development, and discusses emerging preventive and therapeutic strategies targeting arsenic biotransformation pathways.

Patulin (PAT), a prevalent mycotoxin. It is widely present in fruits and nuts and causes serious harm to human health. Our prior study found that PAT exposure could trigger ferroptosis, which in turn resulted in severe kidney damage. However, the specific mechanism remained unclear. This research was designed to explore the molecular mechanisms responsible for PAT-induced renal ferroptosis. In vivo, Western blot analysis revealed that PAT exposure activated ferroptosis and endoplasmic reticulum (ER) stress, and reduced the expression of mitochondria-associated endoplasmic reticulum membranes (MAMs)-associated proteins. Specific commercial detection kits revealed a decrease in tissue adenosine triphosphate (ATP) content and an increase in iron content. Transmission electron microscopy (TEM) observation showed that PAT disrupted the structure of MAMs. In vitro, PAT exposure activated the PERK-related ER stress pathway in mouse kidneys and disrupted both the structure of MAMs and normal mitochondrial function. Molecular docking revealed a strong interaction between MFN2 and MFN1. ER stress inhibition in PAT-treated HKC cells elevated MAMs-related protein expression, which in turn restored mitochondrial membrane potential (MMP) and lowered Mitochondrial Reactive Oxygen Species (MtROS) levels. Similarly, MFN2 overexpression restored mitochondrial function and inhibited ferroptosis. Our study demonstrated that ER stress and MAMs integrity lay at the heart of PAT-induced mitochondrial failure and ferroptosis. The results identified a new mechanism through which PAT triggered ferroptosis, offering fresh perspectives on the pathogenesis of mycotoxin-induced kidney injury.

Patulin (PAT) is a common mycotoxin widely found in various agricultural products and fruits, which has obvious toxic effects on animals and humans. Some studies have shown that PAT can cause nephrotoxicity, but the exact mechanism remains to be elucidated. In the present study, we investigated PAT-induced nephrotoxicity and the possible molecular mechanisms involved in its action. In vivo, the results showed that PAT affected the integrity of the glomerular basement membrane and peduncles, leading to necroptosis. We further demonstrated that PAT up-regulated the expression of JAK2, STAT3, RIPK1, RIPK3, and MLKL. This observation was also confirmed in MPC-5 cells. In vitro, pretreatment with Nec-1 (a specific inhibitor of necroptosis) or si-STAT3 resulted in a significant reduction in necroptosis and improved mitochondrial dysfunction. Notably, the pharmacological protection of mitochondrial function by SS-31 significantly attenuated the onset of PAT-induced necroptosis. Taken together, our study suggested that STAT3 activation, and mitochondrial dysfunction played critical roles in PAT-induced necroptosis in the kidney. These findings revealed the mechanisms by which PAT triggered necroptosis, potentially providing a new therapeutic strategy for PAT poisoning.

Arsenic is an environmental pollutant that threatens public health and is significantly correlated with the risk of developing type 2 diabetes (T2D). Sodium arsenate (NaAsO2) exposure can cause lipid peroxidation, leading to hepatic insulin resistance (IR) and ferroptosis. However, the specific mechanism remains unclear. We performed an oxidative lipidomics analysis of liver tissue from rats with NaAsO2-induced IR by mass spectrometry. Differential enrichment most often involved arachidonic acid (AA) metabolites. However, whether CYP450-epoxyeicosatrienoic acid (EET)-mediated AA metabolism influenced arsenic-induced ferroptosis and hepatic IR remains unknown. We found that NaAsO2 inhibited CYP2C18-EET-mediated AA metabolism that led to an increase of reactive oxygen species (ROS) that promoted ferroptosis. In addition, CYP2C18 and EETs reversed NaAsO2-impaired hepatic insulin sensitivity. In conclusion, NaAsO2 suppressed CYP2C18-EETs mediated AA metabolism and induced ferroptosis in rats with hepatic IR. The study results increase our understanding of potential targets for the prevention and treatment of diabetes.

Sodium arsenite (NaAsO2), the most common form of inorganic arsenic prevalent in the environment, has been closely linked to islet β-cell dysfunction, a critical pathological hallmark of type 2 diabetes (T2D). Even though apoptosis plays a pivotal role in arsenic-induced islet β-cell dysfunction, the explicit underlying mechanisms remain elusive. Here, we have identified that the SET-Rac1 signaling pathway is instrumental in the apoptosis and dysfunction of islet β-cells induced by NaAsO2. During NaAsO2-induced islet β-cell apoptosis and dysfunction, our observations indicated downregulation of SET (almost 0.5-fold) and upregulation of Rac1 (0.5-fold). Notably, overexpression of SET or inhibition of Rac1 substantially mitigated the apoptosis of islet β-cells and ameliorated the impaired insulin secretion (increased from 0.1 ng/ml to 0.2 ng/ml) caused by NaAsO2 exposure. In addition, we detected cytoskeletal disorganization following NaAsO2 treatment, characterized by elevated Cofilin-1 protein expression (approximately 2.5-fold) and disrupted cytoskeleton arrangement. Significantly, overexpression of SET or deletion of Rac1 rectified the NaAsO2-induced cytoskeletal abnormalities, as evidenced by the reduced Cofilin-1 expression and enhanced F-actin fluorescence. Our research delineates that NaAsO2 triggers apoptosis and functional impairment of islet β-cells through cytoskeletal rearrangement mediated by the SET-Rac1 pathway. This discovery could provide novel insights into therapeutic strategies for T2D provoked by environmental toxicants.

Arsenic, recognized as an environmental and food contaminant, has been linked to the dysfunction of islet β-cells, the primary lesions in type 2 diabetes (T2D). Ferroptosis, a regulated cell death pathway dependent on GPX4, has been implicated in arsenic-induced β-cell dysfunction. However, the underlying molecular mechanisms remain unclear. GPX4 activity is significantly modulated by glutathione levels. In this study, we demonstrate that arsenic inhibits GPX4 expression by upregulating the expression of glutathione-specific γ-glutamylcyclotransferase 1 (CHAC1) (>2-fold in vivo and 1.5-fold in vitro). Conversely, arsenic does not affect the expression of the glutathione-cysteine ligase catalytic subunit (GCLC), which is crucial for glutathione synthesis. Notably, CHAC1 knockdown significantly ameliorated arsenic-induced β-cell dysfunction and ferroptosis. N6-methyladenosine (m6A) plays a crucial role in the post-transcriptional modification of mRNA. Arsenic treatment downregulated the expression of methyltransferases METTL3/14 (approximately 0.5-fold), and overexpression of METTL3 alleviated arsenic-induced β-cell dysfunction and ferroptosis. The m6A modification site on CHAC1 was identified, and RIP assays confirmed that arsenic treatment inhibited the interaction between METTL3/YTHDF2 and CHAC1. Furthermore, METTL3 overexpression reduced the half-life of CHAC1 mRNA (almost 0.5-fold). This study uncovers a novel mechanism by which arsenic modulates CHAC1 and ferroptosis through m6A in β-cell dysfunction, highlighting potential therapeutic targets for arsenic-related T2D.

Arsenic in the environment, such as sodium arsenic (NaAsO2), is a frequently occurring hazard that has been linked to nonalcoholic steatohepatitis (NASH). Our prior research established the involvement of ferroptosis in arsenic-induced NASH, but the precise underlying mechanisms remain elusive. Here, we found that exposure to NaAsO2 had a suppressive effect on the expression of CDGSH iron-sulfur domain-containing protein 2 (CISD2) at the protein and gene levels, and overexpression of CISD2 inhibited NaAsO2-induced ferroptosis and NASH. Additionally, administration of NaAsO2 to hepatocytes triggered mitochondrial dysfunction, manifesting as the release of cytochrome c, impairment of the mitochondrial respiratory chain, and reduction in ATP synthesis. However, these adverse effects were alleviated through overexpression of CISD2. Intracellular iron redistribution was induced by overexpression of CISD2 and inhibited NaAsO2-induced ferroptosis. This inhibition was characterized by a reduction in cytoplasmic iron levels and an increase in mitochondrial iron levels. Our study demonstrated that NaAsO2 induced intracellular iron reorganization and mitochondrial dysfunction through CISD2 inhibition, leading to ferroptosis and NASH. This may provide a novel means of treatment of nonalcoholic fatty liver disease triggered by environmental factors.

Di(2-ethylhexyl) phthalate (DEHP), the most prevalent plasticizer worldwide, can enter the human body via various exposure pathways, including ingestion, inhalation, skin contact, and medical applications. It has been observed to exhibit characteristic patterns of accumulation in the female reproductive system. Research has demonstrated that DEHP poses a threat to reproductive health in females across various life stages. Exposure during childhood has been demonstrated to induce central precocious puberty (CPP). Exposure during reproductive age has been shown to be closely associated with reduced fertility, reproductive system tumors, polycystic ovary syndrome (PCOS), and increased risk of endometriosis. Furthermore, exposure during the perinatal period has been demonstrated to increase the risk of embryo implantation failure, placental dysfunction, and abnormal ovarian development in offspring. The toxic effects of DEHP exhibit transgenerational transmission characteristics, with its metabolic products' endocrine-disrupting activity and oxidative stress-inducing capacity being the core toxic factors. This study integrates toxicological evidence linking DEHP exposure to female reproductive damage, providing important references for environmental health risk assessment and the prevention and control of female reproductive diseases.

The mechanism for induction of ferroptosis and islet β-cell dysfunction following arsenic exposure remains elusive. Solute carrier family 25 member 12 (SLC25A12) is a component of the malate-aspartate shuttle and is essential for insulin secretion. However, the role of SLC25A12 in arsenic-induced ferroptosis and islet β-cell dysfunction is unknown. Because ferroptosis is involved in pancreatic dysfunction through mitochondrial reactive oxygen species (MtROS), the present study sought to elucidate the explicit mechanism of MtROS generation in arsenic-induced ferroptosis and pancreatic dysfunction. SLC25A12 was identified as a key mediator of arsenic-induced islet β-cell dysfunction and ferroptosis. Mechanistically, SLC25A12 suppression inhibited glutamate transport from the cytoplasm to mitochondria and decreased MtROS levels by alleviating mitochondrial dysfunction. Thus, SLC25A12-mediated glutamate translocation and MtROS generation contributed to NaAsO2-triggered islet β-cell dysfunction and ferroptosis. These results provided new clues for the SLC25A12 potential as a therapeutic target for diabetes.

The persistent organic pollutant perfluorooctane sulfonate (PFOS) is demonstrated to induce hepatotoxicity through disrupting iron homeostasis and subsequent ferroptosis in hepatocytes. However, it is still elusive in the mechanisms underneath the dysfunctional iron metabolism caused by PFOS. In this study, we observed that PFOS activated the nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in mice liver and human hepatocytes. PFOS reduced the ubiquitination of NCOA4, subsequently causing an increase in the expression of NCOA4. PFOS induced the ubiquitination of HECT and RLD domain-containing E3 ubiquitin protein ligase 2 (HERC2), an upstream negative regulator of NCOA4, leading to the degradation of HERC2. PFOS upregulated the level of detyrosinated α-tubulin (detyr-α-tubulin) in hepatocytes. Under PFOS exposure, detyr-α-tubulin interacted with tripartite motif containing 21 (TRIM21), another E3 ubiquitin ligase responsible for HERC2 degradation. Despite the reduction in the protein level of HERC2, the increases in detyr-α-tubulin and the interaction between detyr-α-tubulin and TRIM21 caused by PFOS facilitated the interaction between TRIM21 and HERC2. Furthermore, inhibiting α-tubulin detyrosination by parthenolide reversed the ferritinophagy and the following ferroptosis caused by PFOS. Collectively, this study points out the existence of ferritinophagy and enriches the understanding of the alteration in iron metabolism under PFOS exposure, providing novel mechanistic insights into the hepatic toxicity of PFOS.

Perfluorooctane sulfonate (PFOS), a listed persistent organic pollutant, poses risks to human health and is closely linked to chronic metabolic diseases. Although the role of mitochondrial fission in these diseases has garnered attention, whether and how PFOS induces mitochondrial fission remains obscure. Here, we found that PFOS induced mitochondrial fission, as demonstrated by the fragmentation of mitochondria and the upregulation of dynamin-related protein 1 (DRP1), phospho-DRP1 and mitochondrial fission protein 1 (FIS1) in human hepatocytes MIHA and mice liver. Blocking the calcium transfer from lysosomes to mitochondria that was executed by transient receptor potential mucolipin 1 (TRPML1) of lysosomes and voltage-dependent anion channel 1 (VDAC1) of mitochondria, did not affect PFOS-induced mitochondrial fission. In contrast, knockdown of TRPML1 or VDAC1 reversed this process. Knockdown of mitochondrial calcium uniporter (MCU), rather than inhibiting its activity, effectively alleviated PFOS-induced mitochondrial fission. Additionally, PFOS increased MCU oligomers without affecting MCU monomer. Inhibiting autophagy reversed the MCU oligomerization. Further investigation unveiled the interactions of MCU with VDAC1, TRPML1, mitochondrial Fo complex subunit F2 (ATP5J2) and DRP1 in PFOS-exposed mice liver and MIHA cells. We also discovered that knockdown of ATP5J2 alleviated PFOS-induced mitochondrial fission. Ulteriorly, PFOS upregulated ATP5J2 that underwent oligomerization. Knockdown of MCU reversed the increase in ATP5J2. Our study uncovers the presence and molecular basics of lysosomes-regulated mitochondrial fission under PFOS exposure, explains the regulatory pathways on MCU and ATP5J2 oligomerization and their pivotal roles in mitochondrial fission, highlighting the involvement of mitochondrial fission in PFOS-related health risks.

BACKGROUND/OBJECTIVES:Di-2-ethylhexyl phthalate (DEHP) is a universally used plasticizer and EDCs. Our previous studies verified that prolonged contact with DEHP induced detrimental impacts on reproductive physiology. Resveratrol (RES), a polyphenolic compound predominantly concentrated in Vitis vinifera epidermis, exhibits multifaceted pharmacological properties. The purpose of this study was to investigate whether RES can alleviate testicular injury caused by DEHP, and to investigate its possible mechanism. METHODS:The experimental design comprised 4 randomized groups of male SD rats: Control group: each animal was given corn oil and saline. DEHP group: the animals received DEHP (500 mg/kg/day) and saline. DEHP and RES group: the animals received DEHP (500 mg/kg/day) and RES (60 mg/kg/day). RES group: the animals received corn oil and RES (60 mg/kg/day). RESULTS:In vivo, RES alleviated DEHP-induced testicular ferroptosis by upregulating SIRT1 and enhancing its interaction with HIF-1α. This mechanism suppressed mitophagy, decreased Fe²⁺and ROS release, and inhibited lipid peroxidation. TM4 cell experiments confirmed these findings. Notably, SIRT1 knockdown inhibited the remission effect of RES on ferroptosis. CONCLUSIONS:In summary, we demonstrated that RES mitigated testicular damage caused by DEHP by inhibiting ferroptosis through the SIRT1-HIF-1α axis. This investigation delineates novel molecular targets and mechanistic frameworks for developing therapeutic interventions against DEHP toxicity.

Type 2 diabetes (T2D) is a chronic metabolic disease that accounts for more than 90% of diabetic patients. Its main feature is hyperglycemia due to insulin resistance or insulin deficiency. With changes in diet and lifestyle habits, the incidence of T2D in adolescents has burst in recent decades. The deterioration in the exposure to the environmental pollutants further aggravates the prevalence of T2D, and consequently, it imposes a significant economic burden. Therefore, early prevention and symptomatic treatment are essential to prevent diabetic complications. Mitochondrial number and electron transport chain activity are decreased in the patients with T2D. Voltage-Dependent Anion Channel 1 (VDAC1), as a crucial channel protein on the outer membrane of mitochondria, regulates signal transduction between mitochondria and other cellular components, participating in various biological processes. When VDAC1 exists in oligomeric form, it additionally facilitates the entry and exit of macromolecules into and from mitochondria, modulating insulin secretion. We summarize and highlight the interplay between VDAC1 and T2D, especially in the environmental pollutants-related T2D, shed light on the potential therapeutic implications of targeting VDAC1 monomers and oligomers, providing a new possible target for the treatment of T2D.