The role and underlying mechanisms of placental ferroptosis in fetal growth restriction (FGR) induced by environmental stress remain poorly understood. Our population-based study showed elevated ferroptosis levels in all-cause FGR placentae. Environmental stressor cadmium (Cd) was used to generate an FGR mouse model, which exhibited elevated placental ferroptosis. Ferroptosis inhibitor ferrostatin-1 reversed environmental Cd-induced FGR. Targeted oxidized lipidomics identified the peroxisome as a target organelle for prenatal Cd-induced placental ferroptosis. Furthermore, Cd induced excessive activation of PEX5-dependent pexophagy in placentae. By establishing a placental Pex5-knockdown mouse, pexophagy was confirmed to drive environmental Cd-induced placental ferroptosis. Mechanistically, pexophagy drives the degradation of the H2O2-scavenging enzyme and the fatty-acid β-oxidation enzymes, thereby causing lipid peroxidation and placental ferroptosis. Notably, environmental Cd upregulated PEX2, an E3 ligase mediating PEX5 monoubiquitination, thereby driving pexophagy and placental ferroptosis. Furthermore, METTL14-mediated m6A modification enhanced the stability of placental PEX2 mRNA in an ELAVL1-dependent manner under environmental Cd. SAH, a METTL14 inhibitor, alleviated Cd-induced placental pexophagy, ferroptosis, and FGR. High-temperature also decreased GPX4 and increased PEX2, PEX5, and METTL14 in placentae. Overall, our findings uncover a novel m6A-PEX2-PEX5 axis driving pexophagy-dependent placental ferroptosis, offering placental pexophagy as a therapeutic target for FGR and fetal-origin adult diseases.

Male infertility affects approximately one in seven couples worldwide. Prenatal cadmium (Cd) exposure has been shown to affect offspring phenotypes and increase susceptibility to diseases later in life. However, the effects of prenatal Cd exposure on multi-generational offspring fertility and the mechanisms remain unknown. A novel murine multi-generational (F1-F3 offspring) male subfertility model induced by prenatal Cd exposure was developed. The levels of testosterone and steroidogenic enzymes were also lower in these offspring's testes. The ubiquitin-dependent degradation of NR4A1, the upstream transcription factor regulating steroidogenic enzymes, was enhanced across generations upon prenatal Cd exposure. After treatment with MG132, an inhibitor of the ubiquitin-proteasome system, the levels of NR4A1 and steroidogenic enzymes were higher in offspring testes with prenatal Cd exposure. Based on the analysis of the UbiBrowser database and testicular global transcriptome, RAPSN was identified as a novel ubiquitin E3 ligase containing the RING-H2_Rapsyn domain that mediates multi-generational testicular NR4A1 ubiquitination. m6A epitranscriptome analysis revealed that prenatal Cd exposure upregulated RAPSN expression in multi-generational offspring testes, and was attributed to a higher level of m6A modification of Rapsn mRNA. Furthermore, there was a lower level of YTHDC2, a m6A reader, in the multi-generational offspring testes with prenatal Cd exposure. Prenatal and postnatal testicular YTHDC2 overexpression reduced the stability of m6A-methylated Rapsn mRNA to downregulate RAPSN expression in F1-F3 testes. Overall, YTHDC2 reduction-mediated increment in m6A-methylated Rapsn mRNA contributed to prenatal Cd-enhanced multi-generational susceptibility to male subfertility.

Gestational exposure to cadmium (Cd), a widespread environmental toxicant, disrupted placental angiogenesis to induce fetal growth restriction (FGR). This study aimed to clarify the mechanism by which Cd disrupts placental angiogenesis. Human and mouse studies indicated that Cd exposure reduced VEGF-A to disrupt placental angiogenesis. Further data confirmed that gestational Cd exposure promoted estrogen receptor ESR1-specific degradation via ubiquitin-proteasome system (UPS), thereby decreasing placental VEGF-A. Bortezomib, the only clinically approved protease inhibitor, blocked ESR1 degradation to alleviate Cd-impaired placental angiogenesis. Based on mouse and human transcriptomics, WWP2 was identified as an unreported ubiquitin E3 ligase targeting placental ESR1. Specifically, both WWP2 knockdown and its inhibitor NSC2805 treatment consistently reversed environmental Cd-induced placental angiogenesis disorders and FGR. Furthermore, the m6A modification in Wwp2 mRNA was increased in Cd-exposed placentae. METTL3 and ELAVL1 knockdown verified that m6A modification enhanced the stability of Wwp2 mRNA. SAH, an inhibitor for METTL3, not only decreased WWP2 but also alleviated Cd-impaired placental angiogenesis and fetal growth. Based on a human case-control study, m6A-methylated Wwp2 was positively correlated with placental angiogenesis inhibition and all-cause FGR. In conclusion, gestational Cd exposure enhanced m6A modification in Wwp2 mRNA to drive ESR1 degradation, thereby inhibiting placental angiogenesis and fetal growth.

The role of placental cellular senescence in environmental cadmium (Cd)-evoked fetal growth restriction (FGR) and its underlying mechanisms require further clarification. Here, we generated a murine FGR model by simulating internal exposure doses of Cd in humans. Human and mouse studies revealed that placental senescence linked environmental Cd exposure to FGR. Furthermore, environmental Cd degraded mitochondrial anti-aging protein SIRT3 to evoke placental cellular senescence and FGR, as demonstrated by SIRT3 overexpression and its activator resveratrol treatment. Interestingly, CLPP was identified as a mitochondrial protease targeting placental SIRT3 degradation under environmental Cd. In vitro CLPP knockdown and in vivo CLPP inhibitor tamarixetin treatment reversed environmental Cd-induced SIRT3 degradation and placental cellular senescence. Additionally, environmental Cd elevated the level of METTL3 protein to promote m6A modification of ClpP mRNA in placentae. In vitro METTL3 knockdown and in vivo its inhibitor S-Adenosylhomocysteine treatment blocked the activation of CLPP-dependent mitochondrial protease stress, attenuating placental cellular senescence and FGR upon environmental Cd. Based on a human case-control study, m6A-driven CLPP-dependent mitochondrial protease stress was positively correlated with placental cellular senescence and all-cause FGR. Taken together, environmental Cd enhances m6A modification to activate CLPP-dependent mitochondrial protease stress, thereby causing placental cellular senescence and FGR.

The escalating consumption of pharmaceuticals and personal care products (PPCPs) has driven their continuous release into the environment, urgently warranting a comprehensive investigation of aquatic contamination profiles and risks to aquatic ecosystems and humans. This study presents a basin-wide assessment of PPCP occurrence patterns, ecological and human risks in Chaohu Lake Basin, China. Forty priority PPCPs were screened across surface waters (lake and tributaries), water from drinking-water treatment plant, and community tap water. Of the 40 target PPCPs, 17 were identified, with detection rates ranging from 2.5% to 97.5%. Seven PPCPs were found in tap water, with caffeine and DEET each present in 85.7% of samples. Ecological-risk modelling indicated that caffeine and amantadine pose a high risk and moderate risk to algae, while no significant risk to daphnids or fish was identified for any compound. The assessment of human health risks, estimated based on daily water consumption, suggests that current levels of PPCPs do not present a measurable health risk to consumers. Urine samples of elderly residents in areas where drinking water is sourced from Chaohu Lake were collected for PPCPs detection. All urine samples from the elderly contained at least one type of PPCP, the concentration range of PPCPs in urine was from the limit of detection to 1.03 mg/L. Cotinine and DEET posed a potential health risk to 10.87% and 5.3% of all subjects, via urine biomonitoring, respectively.

Recently, the perspective of paternal origin has emerged, yet its role in motor disorders remains unclear. Here, using Drosophila and murine models, we demonstrated that following paternal environmental heavy metal stress, offspring from multiple generations exhibited progressive motor deficits accompanied by dopaminergic neuron loss. Cross-tissue RNA sequencing revealed dysregulation of CG9593 (functionally homologous to human angiopoietin-like protein 4, ANGPTL4), identifying CG9593/ANGPTL4 as a pivotal brain effector linking paternal heavy metal stress to multigenerational motor disorders across species. Specifically, neuron-specific CG9593 knockdown ameliorated these disorders, whereas CG9593 overexpression mimicked them. Mechanistically, N6-methyladenosine (m6A) hypermethylation in sperm stabilized CG9593 messenger RNA in offspring neurons via IGF2BP1-dependent posttranscriptional regulation, reducing synapse-associated protein DLG1 and impairing motor neuron. Human cohort analyses confirmed that serum ANGPTL4 negatively correlated with pediatric motor scores, and paternal sperm m6A levels were positively associated with the heavy metal exposure burden. Our findings establish CG9593/ANGPTL4 as an evolutionarily conserved determinant of paternally acquired multigenerational motor disorders and hold promise as a druggable target.

This study explored the spatiotemporal distributions of microorganisms and antibiotic resistance genes (ARGs) in the surface water, sediments, and fish intestinal contents (IC) of Chaohu Lake, and further revealed the pharmaceuticals and personal care products (PPCPs), pesticides in lake water and their relationships with ARGs. 53 types of pesticides and 25 types of PPCPs were identified in the river-lake system basin, with the highest concentrations observed for tebuconazole (1142.36 ng/L) and amantadine (851.41 ng/L). Higher concentrations of these target contaminants were detected in the western part of the lake and during the wet season. No significant east-west differences in ARGs and microbiota across the three environmental matrices were found, whereas seasonal variations impacted lake water and IC more profoundly than sediments. Significant correlations between microbiota and ARGs in the lake water and IC, while those in sediments showed weaker associations. Co-occurrence network analysis showed, among three environmental media, β-lactam and multidrug-resistant genes were widespread found, and Proteobacteria were identified as a stable host. Non-antibiotic pollutants, such as amantadine and terbuthylazine, displayed significant positive correlations with ARGs, similar to sulfamethoxazole and clindamycin. Our findings highlight the important roles of seasonal hydrological changes and non-antibiotic pollutants in the dissemination of ARGs.

Chaohu Lake, a vital freshwater resource for surrounding communities, faces environmental stress due to industrial and mining activities, leading to metal contamination. However, the potential ecotoxicological effects of these metals remain unclear. To address this knowledge gap, we analyzed spatial metal distributions in the lake and developed a Drosophila model exposed to region-specific raw water samples. Our results revealed significantly elevated total metal levels in the western region compared to other areas. Male Drosophila exposed to western raw water exhibited >20% reduced climbing ability and reproductive deficits, suggesting metals as likely contributors to these adverse effects. Notably, the primary characteristics of the observed reproductive dysfunction included a >20% reduction in motor activity and a more than twofold increase in brain senescence in offspring following paternal exposure to western water. Comparative analysis with China's surface water standards identified barium (Ba), molybdenum (Mo), and cadmium (Cd) as key metals in the western region. Crucially, paternal exposure to these metals alone recapitulated the offspring's motor decline and brain senescence. We conclude that Ba, Mo, and Cd may be the key metals responsible for Chaohu Lake water-induced paternal motor/reproductive dysfunction and impaired offspring motor performance, with brain senescence potentially mediating the offspring's motor deficits.

Prenatal environmental stress damages fetal testicular development, leading to male infertility. However, the precise mechanisms underlying the impact of gestational environmental stress on fetal testicular development require further investigation. This study demonstrates that gestational environmental stressor cadmium exposure caused placental estradiol synthesis inhibition and fetal testicular dysplasia. Gestational estradiol supplementation restores fetal testicular dysplasia caused by environmental stress-induced placental estradiol synthesis inhibition. Analysis of human placentae and cadmium-stimulated human primary placental trophoblasts confirmed that ER-phagy is associated with the inhibition of estradiol synthesis in placentae. Subsequently, the data reveals that environmental stress significantly activates RTN3L-mediated ER-phagy. RTN3L-deficient cells and placental Rtn3l-specific knockout mice confirm that environmental stress-activated RTN3L-mediated ER-phagy inhibited placental estradiol synthesis. Total N6-methyladenosine level increasing in gestational environmental stress-exposed placentae. METTL3-mediated N6-methyladenosine modification suppression obviously restrains environmental stress-activated RTN3L-dependent ER-phagy. In conclusion, gestational environmental stress activates ER-phagy by increasing placental Rtn3l mRNA N6-methyladenosine modification, inhibiting placental estradiol synthesis, and contributing to fetal testicular dysplasia. The study demonstrates the early prevention and treatment of adult male infertility from the perspective of fetal-derived diseases.

Cadmium (Cd)-induced male infertility and its mechanisms have been widely studied, but systematic construction of an adverse outcome pathway (AOP) network and identification of key AOPs remained unknown. This study integrated network toxicology, single-cell sequencing, NHANES research, animal experiments, and molecular modeling to construct an AOP network and identify the key AOP for Cd-induced male infertility. We constructed an AOP network comprising 9 AOPs and confirmed that PPARα activation leading to testosterone reduction and subsequent male infertility is the key AOP. Single-cell sequencing analysis showed downregulation of testosterone synthesis genes (Cyp11a1) in Cd-exposed Leydig cells, whereas NHANES data showed a threshold effect between blood Cd (about 1.17 μg/L) and serum testosterone, indicating that high Cd exposure can reduce serum testosterone levels. Subnetwork analysis prioritized PPARα activation and increased the number of reactive oxygen species as potential critical MIEs. In vivo experiments further validated that the PPARα inhibitor (GW6471) more effectively reversed Cd-evoked sperm count reduction than the antioxidant N-acetylcysteine. Taken together, these results validated PPARα activation-mediated testosterone reduction as the key pathway for Cd-induced male infertility and established a novel integrative paradigm for identifying key AOPs in environmental toxicology.

Lead is a major heavy metal pollutant resulting from industrial activities, such as mining and battery manufacturing. Residual Pb ions pose serious health risks, highlighting the need for accurate monitoring to ensure environmental safety. Although traditional methods, such as inductively Coupled Plasma Mass Spectrometry (ICP-MS), are reliable, they require costly equipment, skilled operators, and are time-consuming, making them impractical for rapid field detection. Consequently, research is focused on developing simpler, faster, and more cost-effective detection methods for environmental monitoring and food safety. In this study, we developed a novel G-quadruplex-based detection system using bioenzyme-driven DNA molecular machines aiming to address the complexities of environmental detection and the need for timely monitoring. The unique spatial folding characteristics of G-quadruplexes can significantly accelerate amplification cycles, thereby reducing the dependence on bioenzymes, lowering detection costs, and enhancing the system's resistance to interference in complex environments. Specifically, we designed a dual-drive DNA detection system based on Pb²⁺-specific DNAzyme and G-quadruplex embedded fluorescent probes. In the presence of Pb²⁺, this system responded rapidly, inducing multiple amplification reactions to achieve high sensitivity and high selectivity detection of Pb²⁺. This detection method can detect Pb²⁺ as low as 1.062 × 10-4 mg/L, with a detection linear range of 3.793 × 10-5-1.897 × 10-2 mg/L. Additionally, this platform demonstrated excellent Pb²⁺ specificity and stability in complex samples. We also compared this technology with ICP-MS, and the results showed a high consistency between the two methods for the detection of actual samples. Furthermore, we expanded the application of this technology for quantitative detection in lake water, fish, and soil, providing a new solution for the rapid detection of Pb²⁺ in daily environmental and food samples.

Per- and polyfluoroalkyl substances (PFAS) are widespread persistent pollutants and pose a risk to human health. However, the transfer efficiencies (TEs) of PFAS from source water into maternal serum, and its metabolic pathways linking PFAS exposure and human disease are remain unclear. Here, we present an integrative study combining multi-water and serum PFAS analysis, lipidomics, and machine learning to decipher TEs and PFAS-metabolism interactions. Analyzing 30 PFAS across source water (N = 18), tap water (N = 18), water during drinking water treatment process (DWTP, N = 12), and serum of pregnant women alongside 297 lipid species, we developed a SHAP (Shapley Additive Explanations) framework to quantify PFAS origins and serum-specific lipid responses. We found PFAS were prevalent in source water samples, with PFBA, PFPeA, 6:2 FTS, PFOA, and PFOS being predominant chemicals. In addition, DWTP may not be effective in removing PFAS, with most of target chemicals tested exhibiting removal below 50 %. Moreover, source water outperforms tap water in predicting maternal serum PFAS concentrations, suggesting source water contamination reflects maternal exposure more directly. Machine learning further showed that maternal serum lipid metabolism was influenced by PFAS, such as HFPO-DA and N-MeFOSAA mainly disrupted glycerphospholipid homeostasis (e.g., PC and LPC), highlighting risks to maternal metabolism. Our findings pioneer AI-driven tracing of PFAS transfer dynamics and lipidomic disruptions, warranting a comprehensive strategy beyond drinking water to handle water PFAS contamination.

Pollution caused by per- and polyfluoroalkyl substances (PFAS) in surface water has become a global health concern. Nevertheless, due to the continuous production of emerging PFAS, the pollution levels and hazards of several precursors and their metabolites have not been evaluated. In this study, Chaohu Lake was selected as a representative freshwater lake to obtain a deeper understanding of the profiles of emerging PFAS in surface water. Nontarget screening tentatively identified 49 PFAS with a confidence level of ≥L3, which included 12 legacy PFAS and 37 emerging PFAS. Based on a target analysis of 57 PFAS, 18 PFAS were detected, with at least 10 PFAS detected in every water sample, indicating the widespread presence of PFAS in Chaohu Lake. Moreover, a risk-based PFAS priority model was used to prioritize the PFAS in Chaohu Lake. Remarkably, perfluoromethanesulfonic acid (PFMeS) exhibited the highest level of risk index among the intersection PFAS identified by the nontarget screening of Chaohu Lake water and human serum. For validation, the cytotoxicity of PFMeS was further evaluated in vitro. This study considerably expands our understanding of the occurrence, environmental risk, and cytotoxicity of PFAS in Chaohu Lake and also provides an experimentally validated basis for future research on novel contaminants in a water environment.

The blood-testis barrier (BTB), a unique structure established through intercellular connections of Sertoli cells, establishes a protective microenvironment for spermatogenesis and male fertility. Cadmium (Cd), known for its toxicity, is ubiquitously present in the environment. Here, our findings revealed that Cd exposure compromises BTB integrity, as demonstrated by decreased expression of BTB-associated proteins and elevated Dsignal/Dradius values. Mechanistically, we demonstrated that activation of the CXCL2/CXCR2 axis contributes to Cd-induced BTB impairment, as evidenced by experiments using a CXCR2 inhibition model. As key BTB components, Sertoli cells rely on autophagy to maintain their physiological functions. However, the specific role and mechanism of Sertoli cell autophagy in Cd-induced BTB damage remain unknown. Notably, our results showed that autophagy inhibition aggravated the Cd-induced BTB disruption and testicular CXCL2/CXCR2 axis activation in mice, whereas autophagy activation alleviates Cd-evoked BTB disruption and testicular CXCL2/CXCR2 axis activation. Further verification by Sertoli cell specific Atg5 knockout mouse model showed that the autophagy suppression exacerbated Cd-induced BTB disruption and upregulated the expression of CXCL2. Collectively, our finding points out that ATG5-dependent autophagy in Sertoli cells protects against Cd-induced BTB disruption via perturbing CXCL2/CXCR2 axis. Our study not only reveals a novel molecular mechanism underlying Cd-induced reproductive toxicity but also provides potential therapeutic targets for male infertility intervention.

Potentially toxic metals pose threats to ecosystems through bioaccumulation, yet the traceability of metal bioaccumulation pathways and their induction of metabolic disorders in fish tissues remain poorly understood. Here, we present an integrative study combining multi-media metal analysis, lipidomics, and machine learning to decipher metal-lipid interactions in Chaohu Lake, China-a critical freshwater system facing industrial and agricultural pollution. Analyzing 16 metals across water, sediments, and four fish tissues (muscle, liver, gonad, brain) and profiled 297 lipid species specifically in these fish tissues, we developed a SHAP-BF (Shapley Additive Explanations-Bioaccumulation Factor) framework to quantify metal origins and tissue-specific lipid responses. Our framework revealed dual water-sediment sources for brain/gonad metals (e.g., Mn, Ni) versus predominant water-derived accumulation in muscle/liver (Ni, Be, Mn, Co). Lipid profiling identified glyceride dominance in muscle/liver/gonad (64.7-69.1 %) and phospholipid enrichment in brain (75.4 %), with muscle lipids exhibiting the highest sensitivity to aqueous metals. Machine learning further linked Ni and Mn to disrupted phospholipid homeostasis (e.g., PC 39:6, PE 32:1 et al.) in muscle, highlighting risks to nutritional quality and food safety. This work pioneers an AI-driven approach for tracing metal transfer dynamics and lipidomic disruptions, offering a transformative framework for ecological risk assessment and pollution mitigation strategies.

The hybridization chain reaction(HCR) is a widely used nucleic acid amplification technique that is essential for gene expression analysis and disease diagnosis.Despite its inherent stability,traditional HCR often suffers from low detection efficiency,which necessitates the use of supplementary molecular technologies to enhance its performance for detecting trace samples.Drawing insights from confinement theory,we have proposed a novel cruciform DNA scaffold-based HCR reaction system(C-HCR).In our approach,the DNA cruciform is assembled with the initiating probes of the HCR reaction through specific binding between adenine-rich and thymine-rich regions.This assembly with derived chains does not interfere with the HCR process.When a target is present,traditional HCR reactions yield long-chain linear products.However,the incorporation of the DNA cruciform in C-HCR leads to the formation of network-like products,which create favorable conditions for efficient molecular collisions and,in turn,promote high detection efficiency within the system.We utilized miR-21 and miR-27a as model targets to validate our design concept.Our results revealed that the miRNA-specific HCR system for miR-21 and miR-27a achieved significant increases in detection efficiency of 27.8% and 50%,respectively,demonstrating the feasibility and versatility of our design.This study offers a new strategy for enzyme-free amplification systems.

Maternal exposure to glucocorticoids has been associated with adverse outcomes in offspring. However, the consequences and mechanisms of gestational exposure to prednisone on susceptibility to osteoporosis in the offspring remain unclear. Here, we found that gestational prednisone exposure enhanced susceptibility to osteoporosis in adult mouse offspring. In a further exploration of myogenic mechanisms, results showed that gestational prednisone exposure down-regulated FNDC5/irisin protein expression and activation of OPTN-dependent mitophagy in skeletal muscle of adult offspring. Additional experiments elucidated that activated mitophagy significantly inhibited the expression of FNDC5/irisin in skeletal muscle cells. Likewise, we observed delayed fetal bone development, downregulated FNDC5/irisin expression, and activated mitophagy in fetal skeletal muscle upon gestational prednisone exposure. In addition, an elevated total m6A level was observed in fetal skeletal muscle after gestational prednisone exposure. Finally, gestational supplementation with S-adenosylhomocysteine (SAH), an inhibitor of m6A activity, attenuated mitophagy and restored FNDC5/irisin expression in fetal skeletal muscle, which in turn reversed fetal bone development. Overall, these data indicate that gestational prednisone exposure increases m6A modification, activates mitophagy, and decreases FNDC5/irisin expression in skeletal muscle, thus elevating osteoporosis susceptibility in adult offspring. Our results provide a new perspective on the earlier prevention and treatment of fetal-derived osteoporosis.

The effects and underlying mechanisms of adolescent exposure to combined environmental hazards on cognitive function remain unclear. Here, using a combined exposure model, we found significant cognitive decline, hippocampal neuronal damage, and neuronal senescence in mice exposed to cadmium (Cd) and high-fat diet (HFD) during adolescence. Furthermore, we observed a significant downregulation of Sirtuin 6 (SIRT6) expression in the hippocampi of co-exposed mice. UBCS039, a specific SIRT6 activator, markedly reversed the above adverse effects. Further investigation revealed that co-exposure obviously reduced the levels of La ribonucleoprotein 7 (LARP7), disrupted the interaction between LARP7 and SIRT6, ultimately decreasing SIRT6 expression in mouse hippocampal neuronal cells. Overexpression of Larp7 reversed the combined exposure-induced SIRT6 decrease and senescence in mouse hippocampal neuronal cells. Additionally, the results showed notably elevated levels of Larp7 m6A and YTH domain family protein 2 (YTHDF2) in mouse hippocampal neuronal cells treated with the combined hazards. Ythdf2 short interfering RNA, RNA immunoprecipitation, and RNA stability assays further demonstrated that YTHDF2 mediated the degradation of Larp7 mRNA under combined exposure. Collectively, adolescent co-exposure to Cd and HFD causes hippocampal senescence and cognitive decline in mice by inhibiting LARP7-mediated SIRT6 expression in an m6A-dependent manner.

Neural tube defects (NTDs) represent a prevalent and severe category of congenital anomalies in humans. Cadmium (Cd) is an environmental teratogen known to cause fetal NTDs. However, its underlying mechanisms remain elusive. This study aims to investigate the therapeutic potential of lipophagy in the treatment of NTDs, providing valuable insights for future strategies targeting lipophagy activation as a means to mitigate NTDs.We successfully modeled NTDs by Cd exposure during pregnancy. RNA sequencing was employed to investigate the transcriptomic alterations and functional enrichment of differentially expressed genes in NTD placental tissues. Subsequently, pharmacological/genetic (Atg5-/- placentas) experiments confirmed that inducing placental lipophagy can alleviate Cd induced-NTDs. We found that Cd exposure caused NTDs. Further analyzed transcriptomic data from the placentas with NTDs which revealed significant downregulation of low-density lipoprotein receptor associated protein 1(Lrp1) gene expression responsible for positive regulation of low-density lipoprotein cholesterol (LDL-C) transport. Correspondingly, there was an increase in maternal serum/placenta/amniotic fluid LDL-C content. Subsequently, we have discovered that Cd exposure activated placental lipophagy. Pharmacological/genetic (Atg5-/- placentas) experiments confirmed that inducing placental lipophagy can alleviate Cd induced-NTDs. Furthermore, our findings demonstrate that activation of placental lipophagy effectively counteracts the Cd-induced elevation in LDL-C levels. Lipophagy serves to mitigate Cd-induced NTDs by reducing LDL-C levels within mouse placentas.

Infertility caused by lipopolysaccharide (LPS) exposure due to infection is endangering male fertility worldwide, but the mechanism remains unclear. The blood-testis barrier (BTB) is essential for maintaining spermatogenesis and male fertility. In the present study, we showed that LPS (5.0 mg/kg) treatment markedly down-regulated the expression of BTB-related proteins, expanded the biotin penetration distance and caused histopathological injury in seminiferous tubules in mouse testes. Notably, testicular macrophage M1 polarization induced by LPS seems to be related to BTB damage, which was well confirmed by co-culture of RAW264.7 and TM4 cells in vitro. Interestingly, a low-dose LPS (0.1 mg/kg) pretreatment attenuated down-regulation of BTB-related proteins expression and histopathological injury and shorten biotin penetration distance in seminiferous tubules caused by LPS. Correspondingly, a low-dose LPS pretreatment suppresses testicular macrophage M1 polarization induced by LPS in mouse testes. Further experiments revealed that histone deacetylase 5 (HDAC5) was markedly down-regulated at 2 h and slightly down-regulated at 8 h, but up-regulated at 24 h in mouse testes after LPS treatment. Additionally, low-dose LPS pretreatment against the down-regulation of HDAC5 protein caused by LPS treatment. Notably, the suppressed testicular macrophage M1 polarization by low-dose LPS pretreatment was broken by BRD4354, a specific inhibitor of HDAC5 in vitro. These results suggest suppressed testicular macrophage M1 polarization by HDAC5 enforces insensitivity to LPS-elicited BTB damage.