Early life stage exposure to fenbuconazole causes multigenerational cardiac developmental defects in zebrafish and potential reasons.

With the increasing use of triazole fungicides in agriculture, triazole pesticides have aroused great concern about their toxicity and ecological risk. The current study investigated the impairments of embryonic exposure to fenbuconazole (FBZ) on cardiac transgenerational toxicity and related mechanisms. The fertilized eggs were exposed to 5, 50 and 500 ng/L FBZ for 72 h, and the larvae were then raised to adulthood in clean water. The adult fish were mated with unexposed fish to produce maternal and paternal F1 and F2 embryos, respectively. The results showed that increased arrhythmia were observed in F0, F1 and F2 larvae. Transcriptome sequencing indicated that the pathway of adrenergic signaling in cardiomyocytes was enriched in F0 and F2 larvae. In both F0 and F1 adult zebrafish hearts, ADRB2 protein expression decreased, and transcription of genes related to cardiac development and Ca2+ homeostasis was downregulated. These alterations might cause cardiac developmental defects. Significantly decreased protein levels of H3K9Ac and H3K14Ac might be linked with the downregulation in transcription of cardiac development genes. Protein‒protein interaction analysis exhibited that the pathway affecting the heart was well inherited in the paternal line. These results provide new ideas for the analysis and prevention of congenital heart disease.

Chronic exposure to low levels of phenanthrene induces histological damage and carcinogenic risk in the uterus of female mice.

Phenanthrene (Phe), a polycyclic aromatic hydrocarbon with low molecular weight, is detected in the environment at high frequency. To study the toxic effects of Phe on the uterine structure and function, female Kunming mice were exposed to Phe (0.05, 0.5, 5 ng/mL) for 270 days by drinking water. Pathological alterations and their action pathways were analyzed using immunohistochemical and biomolecular technology. Phe significantly increased the percentage of blood vessel area, the number of endometrial neutrophils (indicating the occurrence of inflammation), collagen deposition (indicating fibrosis), and the percentage of Ki-67-positive cells (indicating carcinogenesis) in the uterus. Transcriptome sequencing identified differentially expressed genes that were mainly enriched in some signaling pathways, including inflammation and carcinogenesis, suggesting a carcinogenic risk in the Phe-exposed uterus. Elevated serum estrogen levels and decreased progesterone levels exhibited a disturbance of steroid hormone balance, which might be related to uterine damage. Upregulated protein levels of uterine androgen receptor and estrogen receptor α were linked to the pathological effects. Most of the effects exhibited a nonmonotonic dose response, which might be attributed to the corresponding change in the serum levels of Phe. The results suggest that exposure to low levels of Phe could exert adverse effects on the uterus.

Prenatal EGCG consumption impacts hepatic glycogen synthesis and lipid metabolism in adult mice.

In this study, the impact of prenatal exposure to Epigallocatechin gallate (EGCG) on the liver of adult offspring mice was investigated. While EGCG is known for its health benefits, its effects of prenatal exposure on the liver remain unclear. Pregnant C57BL/6 J mice were exposed to 1 mg/kg of EGCG for 16 days to assess hepatotoxicity effects of adult offspring. Transcriptomics and metabolomics were employed to elucidate the hepatotoxicity mechanisms. The findings revealed that prenatal EGCG exposure led to a decrease in liver somatic index, enhanced inflammatory responses and disrupted liver function through increased glycogen accumulation in adult mice. The integrated omics analysis revealed significant alterations in key pathways involved in liver glucose lipid metabolism, such as gluconeogenesis, dysregulation of insulin signaling, and induction of liver inflammation. Furthermore, the study found a negative correlation between the promoter methylation levels of Ppara and their mRNA levels, suggesting that EGCG could reduce hepatic lipid content through epigenetic modifications. The findings suggest that prenatal EGCG exposure can have detrimental impacts on the liver among adult individuals and emphasize the need for a comprehensive evaluation of the potential risks associated with EGCG consumption during pregnancy.

The effects of EGCG supplementation on pancreatic islet α and ß cells distribution in adult male mice.

Tea and tea products are widely used as the most popular beverage in the world. EGCG is the most abundant bioactive tea polyphenol in green tea, which has positive effects on the prevention and treatment of diabetes. However, the impact of EGCG exposure on glucose homeostasis and islets in adult mice have not been reported. In this study, we studied glucose homeostasis and the morphological and molecular changes of pancreatic islet α and ß cells in adult male mice after 60 d of exposure to 1 and 10 mg/kg/day EGCG by drinking water. Glucose homeostasis was not affected in both EGCG groups. The expression of pancreatic duodenal homebox1 (Pdx1) in ß cells was upregulated, which might be related to increased insulin level, ß cell mass and ß cell proliferation in 10 mg/kg/day EGCG group. The expression of aristaless-related homeobox (Arx) in α cells did not change significantly, which corresponded with the unchanged α-cell mass. The significant reduction of musculoaponeurotic fibrosarcoma oncogene homolog B (MafB) positive α-cells might be associated with decreased glucagon level in both EGCG groups. These results suggest that EGCG supplementation dose-dependent increases ß cell mass of adult mice and affects the levels of serum insulin and glucagon. Our results show that regular tea drinking in healthy people may have the possibility of preventing diabetes.

Benzo(a)pyrene exposure in early life suppresses spermatogenesis in adult male zebrafish and association with the methylation of germ cell-specific genes.

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants that are widely present in aquatic ecosystems. To assess the impact of early-life exposure to benzo[a]pyrene (BaP), a representative PAH, on reproductive ability in adult male zebrafish (Danio rerio), fertilized embryos were exposed to 0.05, 0.5, 5 and 50 nM of BaP for 96 h, and then the hatched larvae were raised to adulthood in clean water. In one-year-old male fish, the percentage of spermatozoa in testis was significantly reduced in the 0.5, 5 and 50 nM treatments. When the treated fish were mated with untreated fish, significantly decreased rate of egg fertilization and hatching success and significantly elevated malformation rate the F1 larvae were observed in the 0.5, 5 and 50 nM treatments. The transcriptional levels of genes along the brain-pituitary-gonadal axis, involving gnrh3, gnrhr3, fshß, lhß, lhγ, lhrγ and ar, were downregulated. In addition, embryonic BaP exposure upregulated the promotor methylation of germ cell-specific genes in the testis of adult fish. The upregulated methylation of ddx4, dnd1, nanos2 in the testis might be associated with the downregulated mRNA levels of these genes, which could be another reason for the inhibition of spermatogenesis. These results indicate that early-life exposure to BaP suppress the reproductive capability of adult male fish, which would cause a decrease in fish population.

Exposure of embryos to phenanthrene impacts the cardiac development in F1 zebrafish larvae and potential reasons.

To explore the impact of embryonic exposure to phenanthrene (Phe), a typical tricyclic polycyclic aromatic hydrocarbon, on cardiac development in next generation, fertilized zebrafish embryos were exposed to 0.05, 0.5, 5 and 50 nM Phe for 96 h, and then transferred to clear water and raised to adulthood. The cardiac development in F1 larvae generated by adult females or males mated with unexposed zebrafish was assessed. Malformation and dysfunction of the heart, such as increased heart rate, arrhythmia, enlarged heart and abnormal contraction, were shown in both paternal and maternal F1 larvae. A greater impact on the distance between the sinus venosus and bulbus arteriosus was exhibited in maternal F1 larvae, while paternal F1 larvae displayed a more severe impact on heart rate and arrhythmia. The transcription of genes related to cardiac development was disturbed in F1 larvae. DNA methylation levels in the promoter of some genes were associated with their transcription. The expression of acetylated histone H3K9Ac and H3K14Ac in maternal F1 larvae was no significantly changed, but was significantly downregulated in paternal F1 larvae, which might be associated with the downregulated transcription of tbx5. These results indicate that exposure to Phe during embryogenesis adversely affects cardiac development in F1 generation, and the effects and toxic mechanisms showed sex-linked hereditary differences, highlighting the risk of Phe exposure in early life to heart health in next generation.

Prenatal EGCG consumption causes obesity and perturbs glucose homeostasis in adult mice.

Epigallocatechin gallate (EGCG) has a wide consumption for its health advantages. The current study investigates the effects of prenatal EGCG administration on glucose metabolism and obesity in adulthood. Pregnant C57BL/6J mice were supplemented with EGCG in drinking water (3 µg/mL) for 16 d. Abdominal obesity was observed in both male and female adult mice, which was associated with the upregulation of adipose-specific genes, including C/ebpα and Srebf1 (Srebf1 only in males), and the downregulation of genes related to lipolysis, such as Acox1, Atgl and Pdk4 (only in males) in visceral adipose tissue. Elevated fasting glucose levels and hyperinsulinemia were observed in adult males, while females exhibit lower glucose level in glucose tolerance test, which might be due to reduced glucagon levels. Though hepatic expression of the insulin receptor signaling pathway was upregulated in males and was not altered in females, prenatal treatment with EGCG downregulated the expression of this signaling pathway in the skeletal muscle of adult mice, which was further demonstrated in primary human skeletal muscle cells treated with EGCG. The methylation levels in promotor of genes related to the insulin receptor signaling were matched with their transcription in mice, while the expression of acetylated histones was downregulated in human skeletal muscle cells. These results suggest that EGCG consumption during pregnancy should be a risk factor for the disruption of glucose homeostasis in adulthood.

Comparative Toxicity of 3-5 Ringed Polycyclic Aromatic Hydrocarbons to Skeletal Development in Zebrafish Embryos and the Possible Reason.

Polycyclic aromatic hydrocarbons (PAHs) are pervasive pollutants in the environment. To compare the developmental toxicity of PAHs with different ring numbers to fish embryos, benzo(a)pyrene (BaP), pyrene (Pyr) and phenanthrene (Phe) were selected as the representatives of 3, 4 and 5-ringed PAHs, and fertilized embryos of zebrafish (Danio rerio) were exposed to 5 nM PAHs for 72 h. The PAH-treated embryos showed defects in craniofacial cartilage. The order of toxicity to the development of craniofacial cartilage was Phe > Pyr > BaP. The transcription of genes related to the development of craniofacial cartilage was downregulated. The GC-MS/MS detection showed that bioaccumulation of BaP in the exposed embryos was two orders of magnitude lower than that of Phe and Pyr. It is suggested that the more uptake and accumulation of Phe and Pyr could be one of the reasons for their greater toxicity to development in early stage embryos.

Long-term exposure to environmental levels of phenanthrene induces emaciation-thirst disease-like syndromes in female mice.

Phenanthrene (Phe) is a polycyclic aromatic hydrocarbon widely present in foods and drinking water. To explore the detrimental effects of Phe on body metabolism, female Kunming mice were treated with Phe in drinking water at concentrations of 0.05, 0.5 and 5 ng/mL. After exposure for 270 d, the animals exhibited dose-dependent reduced body weight and increased water consumption. The dose-dependent accumulation of Phe in the brain decreased hypothalamic neuron numbers, upregulated hypothalamic expression of anaplastic lymphoma kinase, elevated norepinephrine levels in white adipose tissue (WAT) and further activated lipolysis in WAT, leading to a reduction in fat mass. Brown adipose tissue formation was reduced, accompanied by the inhibition of the bone morphogenetic protein signaling pathway. A simultaneous reduced serum levels of antidiuretic hormone (arginine vasopressin) might be one of the reasons for increased water consumption. The present results indicate an environmental etiology and prevention way for the development of emaciation-thirst disease.

EGCG exposure during pregnancy affects uterine histomorphology in F1 female mice and the underlying mechanisms.

Although epigallocatechin-3-gallate (EGCG), the major polyphenol in green tea, has been shown to have many benefits, the effect of EGCG exposure in utero on adult uterine development is unclear. In this study, pregnant C57BL/6 mice were exposed to 1 mg/kg body weight (bw) EGCG dissolved in drinking water from gestational days 0.5-16.5. A significant decrease in uterine weight was observed in the adult female mice, accompanied by uterine atrophy, inflammation, and fibrosis in the endometrium. Uterine atrophy was attributed to the thinning of the endometrial stromal layer and a significant reduction in endometrial cell proliferation. The expression levels of related proteins in the NF-κB and RAF/MEK/ERK signaling pathways were significantly increased, which might be responsible for the occurrence of inflammation. Activation of the transforming growth factor beta (TGF-ß1)/Smad signaling pathway might be involved in the development of endometrial fibrosis. The changes in the expression of estrogen receptor α, ß (ERα, ERß), progesterone receptor (PGR), and androgen receptor (AR) might lead to changes in the aforementioned signaling pathways. The promoter region methylation level of Esr2 was increased, and the expression of DNMT3A was evaluated. Our study indicates a risk of EGCG intake during pregnancy affecting uterine development in offspring.

Prenatal exposure to a mixture of PAHs causes the dysfunction of islet cells in adult male mice: Association with type 1 diabetes mellitus.

Polycyclic aromatic hydrocarbons (PAHs) have been detected throughout the human body. Whether exposure to PAHs is associated with the incidence of type 1 diabetes mellitus should be investigated. To this end, pregnant mice were exposed to mixed PAHs (5, 50, or 500 µg/kg) once every other day during gestation. The adult male offspring displayed impaired glucose tolerance and reduced serum levels of glucagon and insulin. Immunohistochemical staining revealed increased numbers of apoptotic ß-cells and a reduced ß-cell mass in these males. The downregulated expression of pancreatic estrogen receptor α, androgen receptor, and transcription factor PDX1 was responsible for impacting ß-cell development. The relatively reduced α-cell area was associated with downregulated ARX expression. The transcription of Isn2 and Gcg in pancreatic tissue was downregulated, which indicated that the function of ß-cells and α-cells was impaired. Methylation levels in the Isn2 promotor were significantly elevated in mice prenatally exposed to 500 µg/kg PAHs, which was consistent with the change in its mRNA levels. The number of macrophages infiltrating islets was significantly increased, indicating that prenatal PAH exposure might reduce islet cell numbers in an autoimmune manner. This study shows that prenatal exposure to PAHs may promote the pathogenesis of type 1 diabetes mellitus.

Embryonic exposure to phenanthrene caused developmental defects of craniofacial cartilage in F1 larvae.

As a representative polycyclic aromatic hydrocarbon with low ring numbers, phenanthrene (Phe) is ubiquitously present in the environment. In this study, zebrafish embryos were exposed to Phe at 0.05, 0.5, 5 and 50 nmol/L for 96 h, and then cultured to adulthood in clean water, the developmental defects of craniofacial cartilage were observed in F1 larvae produced by adult males and females mated with untreated fish. Delayed development of craniofacial cartilage, including a shorter and wider Meckel's cartilage and mandibular arch were observed in F1 larvae from adult fish of both sexes. Maternal F1 larvae showed a greater impact on the lower jaw than paternal F1 larvae, this may be connected with greater downregulation of the transcription of genes related to the development of craniofacial cartilage such as runt-related transcription factor 2 (runx2), fibroblast growth factor 8 (fgf8), sonic hedgehog (shh), Indian hedgehog (ihh). Further results indicated that the modification DNA methylation levels in the promotors of gene runx2 and shh in maternal and paternal F1 larvae were inherited from embryonic F0 larvae, and might be linked with the toxicity of craniofacial cartilage in F1 larvae. This study illustrated that embryonic exposure to Phe could induce adverse effects on craniofacial development in F1 offspring, emphasizing the importance of transgenerational toxicology studies in risk assessment.

Embryonic exposure to fenbuconazole inhibits gametogenesis in adult zebrafish by targeting gonads not brain.

Fenbuconazole (FBZ) is widely used in agriculture. The current study was conducted to evaluate the influence of embryonic exposure to FBZ on reproduction in adult zebrafish. Embryos were exposed to 5, 50 and 500 ng/L FBZ for 72 h and then raised in clean water until adulthood. The result showed that the percentage of mature gametes was significantly reduced in adult zebrafish. The fertilization rate and survival rate of F1 embryos were decreased when the exposed fish were mated with untreated fish. The transcription of brain gnrh3, fshß and lhγ in adult fish was upregulated, while the levels of 17ß-estradiol and testosterone were not significantly changed in all treated groups, indicating that the reproduction-related genes in brain was not responsible for the reduced reproductive ability. The downregulated transcription of fshr, lhr, ar and esr2 in the gonads indicated the dysfunction of Sertoli and Leydig cells. Notably, downregulated transcription and upregulated methylation levels of genes related to germ cells were observed in treated F0 larvae and adult gonads. The elevated methylation levels of piwil1 and dnmt6 in the testes and vasa and dazl in the ovary were matched with the alterations in the expression of these genes, suggesting that germ cells are the main targets of FBZ. These results provide new mechanism underlying reproductive toxicity in fish caused by chemicals, and give potential retroactive biomarkers for monitoring reproductive toxic pollutants.

Prenatal EGCG exposure-induced heart mass reduction in adult male mice and underlying mechanisms.

Epigallocatechin-3-gallate (EGCG), which is a major polyphenol in tea, has an unclear effect on cardiac development. In the present study, mice (C57BL/6) were exposed in utero to EGCG dissolved in drinking water (3 µg/ml) for 16 days. A significant decrease in the heart/body weight ratio was observed in adult males but not in adult females. The protein expression levels of TGF-ß1 and its downstream transcription factors SMAD3 and SMAD4 were significantly decreased in male hearts. The PI3K/AKT signaling pathway was inhibited, the expression of proapoptotic proteins, such as BAX, Cleaved Caspase3 and Cleaved Caspase9, was elevated, and the level of antiapoptotic proteins, such as BCL-2, was decreased. A reduced heart/body weight ratio may be associated with the loss of cardiac fibers and an increase in myocardial apoptosis. The cardiac levels of aromatic hydrocarbon receptor and androgen receptor were elevated only in males, which may explain the sexual dimorphism in the effects. The promoter methylation levels of pik3r1, tgf-ß, smad4 were elevated, and those of ahr were reduced, explaining the mechanism underlying the cardiac histological alteration caused by prenatal exposure to EGCG. The results suggest that ingestion of EGCG during pregnancy may be a risk factor for cardiac development in offspring.

In utero exposure to mixed PAHs causes heart mass reduction in adult male mice.

Polycyclic aromatic hydrocarbons (PAHs) are a risk factor for the occurrence of cardiac diseases. The present study was conducted to investigate the influence of prenatal exposure to a mixed PAHs on heart and the underlying mechanism. Pregnant mice were orally administered with a mixture of 8 kinds of PAHs (0, 5, 50, 500 µg/kg body weight) once every 2 days for a total of 8 dosages. The mixed PAHs contained naphthalene, acenaphthylene, phenanthrene, fluoranthene, pyrene, benzo[a]pyrene, dibenzo[a,h]anthracene and benzo[g,h,i]perylene at a weight ratio of 10: 10: 10: 10: 10: 1: 1: 1. The adult males, not females, showed significantly decreased heart/body weight ratio, which was attributed to the loss of cardiac fiber and the increase of cell apoptosis. The protein expression of transforming growth factor ß1 and its downstream transcription factors, Smad3 and Smad4, was significantly downregulated, which caused the loss of cardiac fiber. The downregulated phosphatidylinositol 3-kinase and AKT led to increased expression of caspase3, caspase9, BAX and reduced expression of Bcl-2, which was responsible for the increased cell apoptosis. Different levels of aromatic hydrocarbon receptor and sex hormone receptors between males and females were associated with the distinct effect on heart.

Long-term exposure to environmental levels of phenanthrene disrupts spermatogenesis in male mice.

Phenanthrene (Phe) is a tricyclic polycyclic aromatic hydrocarbon with high bioavailability under natural exposure. However, there are few studies on the reproductive toxicity of Phe in mammals. In this study, male Kunming mice were gavaged once every two days with Phe (5, 50, and 500 ng/kg) for 28 weeks. The accumulation levels of Phe in the testis were dose-dependently increased. Histopathological staining showed that Phe exposure reduced the number of spermatogonia, sperm and Sertoli cells. The percentage of testicular apoptotic cells was significantly increased, which was further verified by the upregulated BAX protein. The expression of the GDNF/PI3K/AKT signaling pathway was downregulated, which might suppress the self-renewal and differentiation of spermatogonial stem cells. Meanwhile, Phe exposure inhibited the expression of Sertoli cell markers (Fshr, WT1, Sox9) and the Leydig cell marker Cyp11a1, indicating damage to the function of Sertoli cells and Leydig cells. Serum estrogen and testicular estrogen receptor alpha were significantly upregulated, while androgen receptor expression was downregulated. These alterations might be responsible for impaired spermatogenesis. This study provides new insights for evaluating the reproductive toxicity and potential mechanisms of Phe in mammals.

Maternal exposure to phenanthrene during gestation disturbs glucose homeostasis in adult mouse offspring.

Epidemiological studies have indicated that polycyclic aromatic hydrocarbons were related to diabetes and insulin resistance. However, studies in mammals on the development of diabetes caused by polycyclic aromatic hydrocarbons are lacking. Pregnant mice were orally exposed to phenanthrene (0, 60 and 600 µg kg-1 body weight) once every 3 days during gestation. In adult mouse offspring, in-utero phenanthrene exposure caused glucose intolerance and decreased insulin levels in females, while caused elevated fasting blood glucose and insulin levels in males. Serum resistin and interleukin-6 levels were elevated in offspring of both sexes. Serum adiponectin levels were decreased in females but increased in males. The insulin receptor signals were upregulated in the liver and downregulated in the skeletal muscle of F1 females, while they were inhibited in both tissues of F1 males. The visceral fat weight and body weight of the treated mice were not increased, suggesting that phenanthrene is not an obesogen, which is supported by the nonsignificant alteration in pparγ transcription in visceral adipose tissue. The transcription of retn in visceral adipose tissue was upregulated in both sexes, and that of adipoq was downregulated in females but upregulated in males, which were matched with the promoter methylation levels of these genes. The results indicated that phenanthrene exposure during gestation could disturb adipocytokine levels via epigenetic modification in adult offspring, and further influence glucose metabolism. These results might be helpful for understanding nonobesogenic pollutant-induced insulin resistance and preventing against diabetes without obesity.

In utero exposure to phenanthrene induced islet cell dysfunction in adult mice: Sex differences in the effects and potential causes.

Epidemiological studies show that the burden of polycyclic aromatic hydrocarbons in human body is associated with the occurrence of insulin resistance and diabetes. In the present study, pregnant mice were exposed to phenanthrene (Phe) at doses of 0, 60 and 600 µg/kg body weight of by gavage once every 3 days. The female F1 mice at 120 days of age showed no change in their fasting glucose levels (FGLs) but exhibited significantly decreased homeostasis model assessment (HOMA) ß-cell (49% and 43%) and significantly downregulated pancreatic proinsulin gene (ins2) transcription. The downregulation of transcription factors, such as PDX1, PAX4 and FGF21, indicated impaired development and function of ß-cells. The significantly reduced α-cell mass in 60 and 600 µg/kg groups, and the significantly downregulated expression of proglucagon gene gcg and ARX in the 600 µg/kg group suggested that the development and function of α-cells had been impacted. The males exhibited significantly increased FGLs (1.14- and 1.15-fold) in Phe exposed treatments and significantly elevated HOMA ß-cell (3.15-fold) in the 600 µg/kg group. Upregulated ins2 transcription and FGF21 protein in male mice prenatally exposed to 600 µg/kg Phe suggested that these animals appeared compensatory enhancement in ß-cell function. The reduced serum estradiol levels and downregulated pancreatic estrogen receptor α and ß were responsible for the dysfunction of ß-cells in the females. In the males, the significantly elevated androgen levels in the 600 µg/kg group might be related to the upregulated ins2 transcription, and the increased expression of pancreatic FGF21 further demonstrated the enhancement of ß-cell potential. The results will be helpful for assessing the risk of developing diabetes in adulthood after prenatal exposure to phenanthrene.

Early-life phenanthrene exposure inhibits reproductive ability in adult zebrafish and the mechanism of action.

Phenanthrene (Phe) is a representative polycyclic aromatic hydrocarbon, and its ubiquity makes the risk assessment of Phe in aquatic ecosystems important. To assess the long-term effects of early-life Phe exposure on fish, the embryos of the model organism, zrbrafish (Danio rerio) were exposed to Phe at 0.05, 0.5, 5 and 50 nmol/L for 96 h and then raised to adulthood in clean water. Gonad development and reproductive functions were investigated in 120 day-old fish. The results showed that the percentage of spermatozoa in males and mature oocytes in females were decreased. The spawned egg numbers and the fertilization rate were reduced when the treated fish were mated with untreated fish. The transcription of genes involved in the brain-pituitary-gonadal axis was downregulated. The levels of both 17ß-estradiol and testosterone were significantly decreased in the 5 and 50 nmol/L groups compared with the control group. The methylation levels in the promotor of gnrh3 (encoding gonadotropin releasing hormone) were significantly elevated in the adult fish in the 5 and 50 nmol/L treatments, which might be associated with the downregulation of gnrh3 transcription. These results suggested that embryonic exposure to Phe can inhibit the reproductive ability of adult fish, which should be adequately emphasized in its risk assessment.

The interference effects of bisphenol A on the synthesis of steroid hormones in human ovarian granulosa cells.

Numerous studies have shown that endocrine-disrupting chemicals are one of the important pathogenic factors in women with polycystic ovary syndrome. Our previous study has revealed that bisphenol A (BPA) can cause steroid hormone imbalance, polycystic ovary, and estrus cycle disorder. In this study, we aimed to explore the effect of BPA, a typical environmental estrogen, on the synthesis of steroid hormones in human ovarian granulosa KGN cells. Exposure of KGN cells to BPA (0.5, 5, 50, and 500 µg/L) resulted in the decrease of progesterone (P), estradiol (E2), and the ratio of estradiol to testosterone (E2/T). BPA affected the expression of genes related to steroid hormone synthesis in KGN cells, including the decreased expression of the steroidogenic acute regulatory protein, ferredoxin, and ferredoxin reductase genes during progesterone synthesis; upregulating the expression of cytochrome p450 oxidoreductase gene associated with E2 and T synthesis; and the downregulated cytochrome P450 family 1 subfamily A member 1 and cytochrome P450 family 1 subfamily B member 1 in E2 degradation. BPA also reduced the expression of stimulatory G proteins (GS) in follicle-stimulating hormone receptor (FSHR)/GS/adenylate cyclase (AC) signaling pathway. In summary, our research has demonstrated that environment-relevant level of BPA exposure leads to steroid hormone synthesis disorder in human ovarian granulosa cells, which might cause the reduction of gene expression in hormone synthesis and the suppression of the FSHR/GS/AC signaling pathway.