Prenatal bisphenol S exposure induces hepatic lipid deposition in male mice offspring through downregulation of adipose-derived exosomal miR-29a-3p.

The increased usage of bisphenol S (BPS) results in wide distribution in pregnant women. In this study, pregnant mice were given multiple-dose BPS during gestation. Results showed that prenatal BPS exposure (50 µg/kg/day) induced increased weight gain, dyslipidemia, higher liver triglyceride (TG), adipocyte hypertrophy, and hepatic lipid deposition in male offspring. Exosomes play important roles in regulating lipid metabolism. Here, serum exosomes and adipose miRNA sequencing of male offspring indicated a remarkable decrease in miR-29a-3p expression. To clarify whether adipocyte-derived exosomes mediate hepatic lipid deposition, exosomes were extracted from BPS-treated adipocytes and co-cultured with hepatocytes. These exosomes could be taken up by hepatocytes and promoted lipid deposition, and notably, exosomal miR-29a-3p was downregulated. Furthermore, miR-29a-3p knockdown in adipocyte-derived exosomes promoted hepatocyte lipid deposition, whereas overexpression led to the opposite effect. Also, the role of miR-29a-3p was demonstrated in hepatocytes by overexpressing or knocking it down. Subsequent studies have shown that miR-29a-3p can promote lipid deposition by directly targeting Col4a1. Taken together, prenatal BPS exposure could lead to lower miR-29a-3p yield in adipocyte-derived exosomes and decrease miR-29a-3p content transported to hepatocytes, which further negatively regulate Col4a1 and promote hepatic lipid deposition. Our findings provided clues to maternal environmental exposure-induced liver metabolic diseases.

Preimplantation triclosan exposure alters uterine receptivity through affecting tight junction protein†.

Triclosan is a broad-spectrum antibacterial agent and widely exists in environmental media and organisms. Triclosan exposure has been reported to have adverse effects on reproduction including embryo implantation disorder. During the embryo implantation window, it is vital that the endometrium develops into a receptive state under the influence of ovarian hormones. However, the effect of triclosan on embryo implantation and endometrial receptivity remains unclear. In the current study, we found a decreased embryo implantation rate, serum estrogen, and progesterone levels in mice exposed to triclosan from gestation days 0.5 to 5.5. Through RNA sequencing (RNA-seq), we identified nearly 800 differentially expressed genes, which were enriched in various pathways, including uterus development, inflammatory response, and immune system processes. Among those enriched pathways, the tight junction pathway is essential for the establishment of the receptive state of the endometrium. Then, genes involved in the tight junction pathway, including Cldn7, Cldn10, and Crb3, were validated by quantitative real-time polymerase chain reaction and the results were consistent with those from RNA-seq. Through immunofluorescence staining and western blotting, we confirmed that the tight junction protein levels of CLDN7 and CRB3 were increased. All these findings suggest that preimplantation triclosan exposure reduces the rate of embryo implantation through upregulating the expression of the tight junction genes and affecting the receptivity of the endometrium. Our data could be used to determine the sensitive time frame for triclosan exposure and offer a new strategy to prevent implantation failure.

Prothioconazole induces cell cycle arrest by up-regulation of EIF4EBP1 in extravillous trophoblast cells.

Prothioconazole (PTC) is a new broad-spectrum triazole antibacterial agent that is being widely used in agriculture. PTC has been linked to a number of reproductive outcomes including embryo implantation disorder; however, the exact mechanism underlying this relationship has yet to be determined. Proper trophoblast proliferation and migration is a prerequisite for successful embryo implantation. To elucidate the underlying molecular perturbations, we detect the effect of PTC on extravillous trophoblast cells proliferation and migration, and investigate its potential mechanisms. Exposure to different concentrations of PTC (0-500 µM) significantly inhibited the cell viability and migration ability (5 µM PTC exposure), and also caused the cell cycle arrest at the lowest dose (1 µM PTC exposure). Transcriptome analysis revealed that PTC exposure disturbed multiple biological processes including cell cycle and apoptosis, consistent with cell phenotype. Specifically, eukaryotic translation initiation factor 4E binding protein 1 (EIF4EBP1, 4E-BP1) was identified as up-regulated in PTC exposure group and knockdown of EIF4EBP1, and attenuated the G1 phase arrest induced by PTC exposure. In summary, our data demonstrated that 4E-BP1 participated in PTC-induced cell cycle arrest in extravillous trophoblast cells by regulating cyclin D1. These findings shed light on the potential adverse effect of PTC exposure on the embryo implantation.

Exploration of the developmental toxicity of TCS and PFOS to zebrafish embryos by whole-genome gene expression analyses.

Triclosan (TCS) and perfluorooctane sulfonate (PFOS) are known to have both endocrine disrupting and developmental toxicity effects on zebrafish embryos. Currently, potential molecular mechanisms underlying these toxicological phenomena require further studies. To address this gap in the literature, we used whole transcriptome microarrays to being to address the potential molecular mechanisms underlying developmental toxicity of TCS and PFOS on zebrafish embryos. Zebrafish embryos were exposed to 300 µg/L TCS and 500 µg/L PFOS from 4 to 120 h post fertilization (hpf). Phenotypically, the hatching rate of zebrafish embryos was significantly reduced after TCS exposure at 72 hpf. Additionally, body length was significantly decreased in the TCS treatment group at 120 hpf. Gene ontology analysis of differentially expressed genes revealed that lipid metabolism, steroid metabolism, and organ development-related biological processes were significantly enriched in TCS- and PFOS-treated zebrafish embryos. Furthermore, signaling network analysis indicated that the steroid biosynthesis process was the most significant biological process disrupted by TCS in 120 hpf zebrafish embryos, while organ development was the most significant biological process disrupted by PFOS exposure. Our findings enhance the understanding of the specific types of embryotoxicity elicited by TCS and PFOS, and also provide information that can be used to inform future mechanistic studies.

Association between mercury exposure and thyroid hormones levels: A meta-analysis.

BACKGROUND: The relationship between mercury (Hg) exposure and thyroid hormones (THs) levels in the general population has been inconclusive. We conducted a random effects model meta-analysis to identify the association between Hg exposure and THs levels in the general population. METHODS: This meta-analysis were performed based on the PECO questions (P = general population; E =1ug/L Hg in blood and urine; C =1ug/L incremental increase on; and O = variation of THs levels). We searched four electronic databases including PubMed, Web of Science, Embase, and Cochrane Library for studies published on or before 20th July 2020. Prospective and cross-sectional studies that evaluated the association between Hg exposure and the levels of thyroid stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), free triiodothyronine (FT3), and free thyroxine (FT4) were included. We also assessed aggregate risks for the reliability of the included studies. RESULTS: Initially, we retrieved 4889 articles. 18 studies met our inclusion criteria after screening and 13 articles were eligible to be included in the meta-analysis. The meta-analysis results suggest that blood Hg may be significantly associated with THs levels. The presence of Hg in blood may significantly increase the levels of TSH (ß=0.55; 95%CI: 0.20,0.90; p < 0.001) and FT4 (ß=0.47; 95%CI: 0.11,0.82; p < 0.001), with the opposite association in T4 (ß=-0.02; 95%CI: 0.02, -0.01; p < 0.001). For the subgroup analysis, blood Hg was positively correlated with TSH levels in children and adolescents (ß=0.62; 95%CI: 0.09, 1.15; p < 0.001) and FT4 levels in pregnant women (ß=1.00; 95%CI: 0.99, 1.00; p < 0.001) respectively. CONCLUSIONS: This meta-analysis indicates that exposure to Hg in blood could significantly corrrelate with the levels of TSH, T4, and FT4 in the general population. Therefore, it is crucial to control the use of Hg and strengthen protection of the thyroid.

Development of HSPA1A promoter-driven luciferase reporter gene assays in human cells for assessing the oxidative damage induced by silver nanoparticles.

The exponential increase in the total number of engineered nanoparticles in consumer products requires novel tools for rapid and cost-effective toxicology screening. In order to assess the oxidative damage induced by nanoparticles, toxicity test systems based on a human HSPA1A promoter-driven luciferase reporter in HepG2, LO2, A549, and HBE cells were established. After treated with heat shock and a group of silver nanoparticles (AgNPs) with different primary particle sizes, the cell viability, oxidative damage, and luciferase activity were determined. The time-dependent Ag(+) ions release from AgNPs in cell medium was also evaluated. Our results showed that heat shock produced a strong time-dependent induction of relative luciferase activity in the four luciferase reporter cells. Surprisingly, at 4h of recovery, the relative luciferase activity was >98× the control level in HepG2-luciferase cells. Exposure to different sizes of AgNPs resulted in activation of the HSPA1A promoter in a dose-dependent manner, even at low cytotoxic or non-cytotoxic doses. The smaller (5nm) AgNPs were more potent in luciferase induction than the larger (50 and 75nm) AgNPs. These results were generally in accordance with the oxidative damage indicated by malondialdehyde concentration, reactive oxygen species induction and glutathione depletion, and Ag(+) ions release in cell medium. Compared with the other three luciferase reporter cells, the luciferase signal in HepG2-luciferase cells is obviously more sensitive and stable. We conclude that the luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the oxidative damage induced by AgNPs.

Comparative oxidative stress elicited by nanosilver in stable HSPA1A promoter-driven luciferase reporter HepG2 and A549 cells.

The use of silver nanoparticles (AgNPs) in consumer products and medical applications is growing rapidly owing to their noted antimicrobial properties. As a consequence, a rapid and reliable toxicology test that allows multiple nanosilver screening in a stress responsive pathway-based assay is needed to ensure consumer safety. Here, novel HSPA1A promoter-driven luciferase reporter gene assays based on two cell models widely used in toxicology testing, HepG2 and A549 cell lines, were developed to analyze the oxidative stress elicited by AgNPs. After AgNP exposure, the luciferase activity, cell viability, and oxidative damage in HepG2- and A549-luciferase cells were determined. The uptake and intracellular localization of AgNPs were measured by transmission electron microscopy. The cellular AgNPs were determined by inductively coupled plasma mass spectrometry. Our results showed that both reporter cells were able to take up nanoparticles and furthermore, exposure to AgNPs resulted in an obviously dose-dependent increase in transcriptional activation of the HSPA1A promoter indicated by luciferase activity. Surprisingly, luciferase activity was greater than 200× the control level in HepG2-luciferase reporter cells treated with 50 µg mL-1 AgNPs. These results were in line with the positive responses in cytotoxicity, intracellular reactive oxygen species induction, glutathione depletion, malondialdehyde concentration, and the cellular silver content. Compared with the A549-luciferase cells, the luciferase signal in the metabolic competent HepG2-luciferase cells is obviously more sensitive and stable. We conclude that these two reporter gene cell systems, especially the HepG2-luciferase cells, demonstrated their utilities in predictive screening of the oxidative stress elicited by nanosilver.