Phenanthrene Exposure Research Articles

Interactions between phenanthrene exposure and historical chemical stress: Implications for fitness and ecological resilience of the sentinel species Daphnia magna

Polycyclic aromatic hydrocarbons (PAHs) arise from incomplete combustion of oil, coal, and gasoline, with lipophilic properties facilitating their widespread distribution and persistence. Due to their biochemical attributes, PAHs can accumulate in animal tissues, potentially causing mutagenic and carcinogenic effects. Since the industrial revolution, PAH concentrations in the environment have risen, with lakes showing levels from 0.159 to 33,090 μg/kg sediment. Despite acute toxicity studies showing adverse effects on freshwater organisms, the long-term impacts and synergistic interactions with other pollutants remain largely unexplored.This study investigates the impact of phenanthrene (PHE), a prominent PAH found in aquatic environments, on Daphnia magna, a species of significant ecological importance in freshwater ecosystems globally, being both a sentinel species for chemical pollution and a keystone organism in freshwater aquatic ecosystems. Leveraging the dormancy of D. magna, which spans decades or even centuries, we exposed strains with diverse histories of chemical contaminant exposure to environmentally relevant concentrations of PHE. Initially, acute exposure experiments were conducted in accordance with OECD guidelines across 16 Daphnia strains, revealing substantial variation in acute toxic responses, with strains naïve to chemical pollutants showing the lowest toxicity. Utilizing the median effect concentration EC10 derived from acute exposures, we assessed the impacts of chronic PHE exposure on life history traits and ecological endpoints of the 16 strains. To elucidate how historical exposure to other environmental stressors may modulate the toxicity of PHE, temporal populations of D. magna resurrected from a lake with a well-documented century-spanning history of environmental impact were utilized. Our findings demonstrate that PHE exposure induces developmental failure, delays sexual maturation, and reduces adult size in Daphnia. Populations of Daphnia historically exposed to chemical stress exhibited significantly greater fitness impacts compared to naïve populations. This study provides crucial insights into the augmented effects of PAHs interacting with other environmental stressors.

Maternal exposure to phenanthrene induces testicular apoptosis and Sertoli cell dysfunction in F1 adult male mice: a histological and molecular study.

Phenanthrene, a polycyclic aromatic hydrocarbon, is found in abundance in environmental pollutants, food, and drinking water. This substance can accumulate in body tissues and exert harmful effects. Moreover, phenanthrene can cross the placental barrier, potentially impacting fetal development. We aimed to explore the impacts of maternal exposure to phenanthrene on testicular tissue and Sertoli cell function in F1 mice. Female rats with vaginal plugs were randomly assigned to one of three groups: control, sham, or phenanthrene. The control group received no intervention during pregnancy. In the sham and phenanthrene groups, corn oil and a phenanthrene solution, respectively, were administered via gavage once every 2 days. Offspring were separated by sex 21 days after birth. At 56 days postnatal, male F1 offspring were euthanized, and their testes were harvested for histological and molecular analyses. Phenanthrene exposure was associated with a lower testicular weight and volume, a smaller diameter of the seminiferous tubules, and a relative thinning of the germinal epithelium. These changes were associated with increased cellular apoptosis, as shown by the upregulation of caspase 3 expression. Additionally, we observed an increase in vacuolization and residual bodies within the tissue. Conversely, the number of Sertoli cells and expression levels of Sox9, as well as the Ocln and Itgb1 genes, were found to be lowered. Maternal exposure to phenanthrene impacts both germ cells and Sertoli cells, disrupting their function and leading to fertility disorders in male F1 offspring mice.

Induction of insulin resistance in female mice due to prolonged phenanthrene exposure: Unveiling the low-dose effect and potential mechanisms

Phenanthrene (Phe) is a commonly occurring polycyclic aromatic hydrocarbon (PAH) found in various food sources and drinking water. Previous studies have shown that long-term exposure to Phe in male mice leads to insulin resistance in a dose-dependent manner. However, the effect of Phe on glucose homeostasis in female mice remains unknown. To address this knowledge gap, female Kunming mice were exposed to Phe through their drinking water at concentrations of 0.05, 0.5, and 5 ng/mL. After 270 d of exposure, we surprisingly discovered a low-dose effect of Phe on insulin resistance in female mice, which differed from the effect observed in male mice and showed sexual dimorphism. Specifically, insulin resistance was only observed in the 0.05 ng/mL treatment, and this low-dose effect was also reflected in the concentration of Phe in white adipose tissue (WAT). Differences in metabolic enzyme activities in the liver may potentially explain this effect. The observed sexual dimorphism in Phe exposure could be attributed to variations in estrogen (E2) level and estrogen receptor beta (ERβ) expression in WAT. These findings highlight the association between environmental factors and the development of insulin resistance, emphasizing the pathogenic effect of even low doses of Phe. Moreover, sex dependent-effect should be given more attention when studying the toxic effects of environmental pollutants.

Phenanthrene-Induced Cytochrome P450 Genes and Phenanthrene Tolerance Associated with Arabidopsis thaliana CYP75B1 Gene.

Polycyclic aromatic hydrocarbons (PAHs) form an important group of organic pollutants due to their distribution in the environment and their carcinogenic and/or mutagenic effects. In order to identify at the molecular level some of the players in the biodegradation and tolerance response to PAHs in plants, we have phenotyped 32 Arabidopsis thaliana T-DNA mutant lines corresponding to 16 cytochrome P450 (CYP) genes that showed to be differentially expressed under contrasted stress conditions induced by phenanthrene, a 3-ring PAH. This screening has allowed us to identify CYP75B1 (At5g07990) T-DNA mutants as the only ones being sensitive to phenanthrene-induced stress, supporting that CYP75B1 protein is necessary for PAH tolerance. CYP75B1 codes for a 3'flavonol hydroxylase. CYP75B1 gene was heterologously expressed on yeast in order to investigate whether it affects the A. thaliana response to phenanthrene by participating in its metabolization. Heterologously-produced CYP75B1 enzyme shows to be catalytically efficient against its physiological substrates (e.g., naringenin) but unable to metabolize phenanthrene or 9-phenanthrenol. In contrast, CYP75B1 seems rather involved in phenanthrene tolerance as a crucial element by regulating concentration of antioxidants through the production of 3'-hydroxylated flavonoids such as quercetin and cyanidin. In particular, we report a highly increased generation of reactive oxygen species (H2O2 and singlet oxygen) in cyp75b1 mutants compared to control plants in response to phenanthrene treatment. Overall, CYP75B1 shows to play an important role in the response to the deleterious effects of phenanthrene exposure and this is related to oxidative stress sensitivity rather than metabolization.

Maternal exposure to polycyclic aromatic hydrocarbons during pregnancy and timing of pubertal onset in a longitudinal mother–child cohort in rural Bangladesh

BackgroundIn experimental studies, several polycyclic aromatic hydrocarbons (PAHs) have shown endocrine disrupting properties, but very few epidemiological studies have examined their impact on pubertal development and results have been heterogenous. ObjectiveTo explore if maternal PAH exposure during pregnancy was associated with the offspring’s timing of pubertal onset. MethodsWe studied 582 mother-daughter dyads originating from a population-based cohort in a rural setting in Bangladesh. Maternal urinary samples, collected in early pregnancy (on average, gestational week 8), were analyzed for monohydroxylated metabolites of phenanthrene (1-OH-Phe, Σ2-,3-OH-Phe, and 4-OH-Phe), fluorene (Σ2-,3-OH-Flu), and pyrene (1-OH-Pyr) using liquid chromatography with tandem mass spectrometry (LC-MS/MS). The girls were interviewed on two separate occasions concerning date of menarche, as well as breast and pubic hair development according to Tanner. Associations were assessed using Kaplan-Meier analysis and multivariable-adjusted Cox proportional hazards regression or ordered logistic regression. ResultsIn early pregnancy, the mothers’ median urinary concentrations of Σ1-,2-,3-,4-OH-Phe, Σ2-,3-OH-Flu, and 1-OH-Pyr were 3.25 ng/mL, 2.0 ng/mL, and 2.3 ng/mL respectively. At the second follow-up, 78 % of the girls had reached menarche, and the median age of menarche was 12.7 ± 0.81 years. Girls whose mothers belonged to the second and third quintiles of ΣOH-Phe metabolites had a higher rate of menarche, indicating a younger menarcheal age (HR 1.39; 95 % CI 1.04, 1.86, and HR 1.41; 95 % CI 1.05, 1.88, respectively), than girls of mothers in the lowest quintile. This trend was not observed in relation to either breast or pubic hair development. None of the other maternal urinary PAH metabolites or the sum of all thereof in early pregnancy were associated with age at menarche or pubertal stage. ConclusionsIndications of non-monotonic associations of prenatal phenanthrene exposure with the daughters’ age of menarche were found, warranting further investigation.

Proteogenomic Characterization of Pseudomonas veronii SM-20 Growing on Phenanthrene as Only Carbon and Energy Source.

In this study, we conducted an extensive investigation of the biodegradation capabilities and stress response of the newly isolated strain Pseudomonas veronii SM-20 in order, to assess its potential for bioremediation of sites contaminated with polycyclic aromatic hydrocarbons (PAHs). Initially, phenotype microarray technology demonstrated the strain's proficiency in utilizing various carbon sources and its resistance to certain stressors. Genomic analysis has identified numerous genes involved in aromatic hydrocarbon metabolism. Biodegradation assay analyzed the depletion of phenanthrene (PHE) when it was added as a sole carbon and energy source. We found that P. veronii strain SM-20 degraded approximately 25% of PHE over a 30-day period, starting with an initial concentration of 600 µg/mL, while being utilized for growth. The degradation process involved PHE oxidation to an unstable arene oxide and 9,10-phenanthrenequinone, followed by ring-cleavage. Comparative proteomics provided a comprehensive understanding of how the entire proteome responded to PHE exposure, revealing the strain's adaptation in terms of aromatic metabolism, surface properties, and defense mechanism. In conclusion, our findings shed light on the promising attributes of P. veronii SM-20 and offer valuable insights for the use of P. veronii species in environmental restoration efforts targeting PAH-impacted sites.

Low levels of flavonoids in Arabidopsis thaliana during phenanthrene exposure suggest new roles for a flavanone 3-hydroxylase

Polycyclic aromatic hydrocarbons (PAHs) contamination impacts the well-being of fauna, flora, and the environment. PAHs are a group of compounds consisting of two or more benzene rings. Due to their structure, PAHs are very stable, generally non-reactive and resistant to biodegradation; yet these compounds are cytotoxic in biological systems. In plants, PAHs induce oxidative stress and cell death, and inhibit growth. Previous PAH exposure studies from our laboratory showed an up-regulation of the Arabidopsis thaliana gene At3g51240. At3g51240 encodes a flavanone 3-hydroxylase (F3H), an enzyme catalyzing the hydroxylation of flavanones, key intermediates in the flavonoid biosynthesis pathway. Considering that flavonoids may act as antioxidants and protect cells against oxidative stress, we hypothesized that F3H was important in alleviating PAH-induced stress symptoms. Analysis of flavonoid content by high performance liquid chromatography showed low levels of flavonoids in phenanthrene-treated plants. Analysis of At3g51240 loss-of-function mutant plant lines growing on phenanthrene-containing growth media showed reduced phenotypic stress responses. Interestingly, low levels of H2O2 were observed in the mutant plants, suggesting that this putative flavanone 3-hydroxylase may be involved in PAH oxidation, with the consequent H2O2 generation. Furthermore, in vitro assays with purified recombinant 6xHis tagged AT3G51240 showed enzyme activity on the PAH when phenanthrene was used as a substrate. Together, these data suggest that F3H is involved in PAH detoxification in planta with the subsequent accumulation of toxic metabolites. This research also indicates the potentiality of using specialized metabolism enzymes such as oxidoreductase in the genetic engineering of plants to remediate PAH-contaminated soils.

Expeditious profiling of polycyclic aromatic hydrocarbons transport and obstruction mechanisms in crop xylem sap proteins via proteomics and molecular docking

Polycyclic aromatic hydrocarbons (PAHs) pose significant environmental risks due to their toxicity and carcinogenic properties. This research seeks to pinpoint protein targets in crop xylem sap related to PAH contamination and delve into their protein-ligand interactions using computational tools. Proteomic assessment revealed differentially expressed proteins (DEPs), which were subjected to virtual high-throughput screening. Notably, the phenanthrene's influence on xylem sap proteins in maize and wheat was more pronounced than in soybean, with DEPs expression peak at 24 h post-treatment. Maize DEPs were predominantly associated with lipid biosynthesis. Phenanthrene impacted cell membrane hydrophobicity, limiting PAH adsorption and decreasing its concentration in maize xylem sap. Wheat DEPs exhibited an increase in ABC transporters after 24 h of phenanthrene exposure. ABC transporters interacted with stress-responsive proteins like C6TIY1-Co-chaperone p23 and others that either facilitate or inhibit PAH transport, including Indeno[1,2,3-cd]Pyrene and C6TIY1-Co-chaperone protein p23. Both maize and wheat created high-affinity complexes between specific proteins and PAHs, influencing their transport. This study provides insights into the mechanisms of PAH regulation and movement within plant xylem.

Marine pollutant Phenanthrene (PHE) exposure causes immunosuppression of hemocytes in crustacean species, Scylla paramamosain

Phenanthrene (PHE), a representative polycyclic aromatic hydrocarbons (PAHs), is widely found in aquatic environments, which exhibits high toxicity to aquatic organisms and has a substantial impact on overall health. In order to investigate the immunosuppressive effects of PHE exposure on marine crustacean species, the Scylla paramamosain was exposure to different concentrations of PHE, which was 0 μg/L (control group), 0.7 μg/L, 7 μg/L, or 70 μg/L PHE with 35 individuals in every group, respectively. The results showed that the color of hemocytes gradually deepened with increasing PHE concentration, and the total hemocyte count (THC) was activated and increased after PHE exposure analyzed by Flow cytometry. Meanwhile, compared with the control group, cryostat sections of hepatopancreas showed cell infiltration, cell steatosis, eosinophilic masses and vacuolization in PHE groups. The superoxide dismutase (SOD) activity was decreased immensely in PHE exposure groups, meanwhile, the acid phosphatase (ACP) activity and glutathione (GSH) activities were increased after PHE exposure compared with control group. Moreover, the expression profile of Crustin, TLR, MCM7, JAK, caspase, Moyosin and P53 were up-regulated significantly after 7th day PHE exposure in all treatment groups by Q-PCR. Those data illustrated that PHE exposure could inhibit the immune function of mud crab by causing hepatopancreas damage, induce the activity of antioxidant enzymes and the expression of immune genes. These data provide a scientific basis for evaluating the impact of PAH pollution on marine organisms.

Decoding the biological toxicity of phenanthrene on intestinal cells of Eisenia fetida: Effects, toxicity pathways and corresponding mechanisms

Phenanthrene is frequently detected and exists extensively in the soil environment, and its residues inevitably impose a significant threat to soil organisms. Exposure to and toxicity of phenanthrene on earthworms has been extensively studied before, however, the possible mechanisms and related pathways associated with phenanthrene-triggered toxicity at the intestinal cell level remain unclear. Herein, primary intestinal cells isolated from Eisenia fetida (Annelida, Oligochaeta) intestine were used as targeted receptors to probe the molecular mechanisms involved in ROS-mediated damaging effects and the potential pathways of phenanthrene-induced toxicity at cellular and sub-cellular levels. Results indicated that phenanthrene exposure induced oxidative stress by activating intracellular ROS (elevated O2−, H2O2, and OH− content) bursts in E. fetida intestinal cells, causing various oxidative damage effects, including lipid peroxidation (increased MDA content), protein oxidation (enhanced PCO levels), and DNA damage (enhanced 8-OHdG levels). The enzymatic and non-enzymatic strategies in earthworm cells were activated to mitigate these detrimental effects by regulating ROS-mediated pathways involving defense regulation. Also, phenanthrene stress destroyed the cell membrane of E. fetida intestinal cells, resulting in cellular calcium homeostasis disruption and cellular energetic alteration, ultimately causing cytotoxicity and cell apoptosis/death. More importantly, the mitochondrial dysfunction in E. fetida cells was induced by phenanthrene-caused mitochondrial membrane depolarization, which in turn caused un-controlled ROS burst and induced apoptosis through mitochondria-mediated caspase-3 activation and ROS-mediated mitochondrial-dependent pathway. Furthermore, exposure to phenanthrene activated an abnormal mRNA expression profile associated with defense regulation (e.g., Hsp70, MT, CRT, SOD, CAT, and GST genes) in E. fetida intestinal cells, resulting in various cellular dysfunctions and pathological conditions, eventually, apoptotic cell death. Taken together, this study offers valuable insights for probing the toxic effects and underlying mechanisms posed by phenanthrene at the intestinal cell level, and is of great significance to estimate the detrimental side effects of phenanthrene on soil ecological health.

Evaluation of life-history traits in Folsomia candida exposed to combined repeated mild heat shocks with phenanthrene.

Climate change increases the frequency and intensity of extreme weather events. In nature, organisms are often exposed to climatic stressors and contaminants simultaneously, and the effects of contaminants may be modified by climate change and vice versa. Here, the effects of repeated mild heat shocks (0-5 times, 30°C for 6h), alone or combined with phenanthrene (PHE) (80mgkg-1 dry soil), on life-history traits of the springtail Folsomia candida were investigated. The survival, growth, maturation, and reproduction of single juvenile springtails were assessed over a period of 37days. Increasing number of heat bouts or PHE exposure did not have significant negative effects on overall survival at the termination of the experiment, but the interaction between the two stressors led to complex interactions for the dynamics of survival during the test. Neither body growth nor time to first oviposition was influenced by heat or PHE, but a reduction of egg production with increasing number of heat bouts was observed, and there was an interaction between the two stressors. Further, a trade-off between the number of eggs produced and egg size was observed, indicating that females invested the same amount of energy in reproduction despite exposure to stressful temperature and PHE. These results indicate that egg production (in terms of the number of eggs) was a more sensitive indicator of the combined effects of mild heat shocks and PHE than growth, and there was a trade-off between survival and egg production.

Effects of Phenanthrene Exposure on the B-esterases Activities of Octopus maya (Voss and Solís Ramírez, 1996) Embryos

No ecotoxicological information exists on phenanthrene (Phe) exposure in cephalopods, animals of commercial and ecological importance. This study investigated the effect of Phe on two B-esterases, Acetylcholinesterase (AChE) and Carboxylesterases (CbE), in Octopus maya embryos. Octopus embryos were exposed to different treatments: control (seawater), solvent control (seawater and DMSO 0.01%), 10 and 100 µg/L of Phe. AChE and CbE activities were measured at different developmental stages (blastula, organogenesis, and growth). B-esterase activities increased in control and solvent control as the embryos developed, showing no statistically significant differences between them. On the other hand, the embryos exposed to Phe had significant differences from controls, and between the high and low concentrations. Our results indicate that B-esterases are sensitive biomarkers of exposure to Phe in O. maya. Still, complementary studies are needed to unravel the toxicodynamics of Phe and the implications of the found inhibitory effect in hatched organisms.

Constructing model-averaging species sensitivity distributions of Phenanthrene based on reproductive fitness: Implications for assessing ecological risk in urban watershed.

The challenge in optimizing the method of constructing species sensitivity distribution (SSD) remains. In this study, a model-averaging SSD was created to evaluate the ecological risk of Phenanthrene (PHE) in urban watershed based on reproductive fitness. Specifically, concentrations of PHE were measured in surface water samples collected from various watersheds of Wuhan, including five lake watersheds and the Wuhan reach of the Yangtze River and Han River. The reproductive endpoint of aquatic species was calculated to be most sensitive to PHE exposure, with the value of predict no-effect concentration (PNEC) at 0.19μg/L. The results of probabilistic assessment methods, including joint probability curve (JPC), overall risk probability (ORP), and distribution-based quotient (DBQ), indicated that the ecological risks of PHE in large lakes have dropped significantly with distance from the downtown area of Wuhan, and the long-term effects of industrial activities may increase the risks in the lake watersheds. Basically, the ecological risks in Yangtze River are negligible; however, there is a relatively high risk of PHE in the Han River and some lake watersheds. The cosθ similarity analysis indicated the Yangtze River is strongly connected to the low-risk lake watersheds, and that in part reflects the risk in the Yangtze River being controlled by its surrounding these lake watersheds.

Response of methanogenic system to long-term polycyclic aromatic hydrocarbon exposure: Adsorption and biodegradation, performance variation, and microbial function assessment

Phenanthrene (PHE) as a typical polycyclic aromatic hydrocarbon (PAH) is prevalent and harmful to organisms in petroleum-polluted sites. The effects of PHE concentration levels on performance, microbial community and functions in methanogenic system were comprehensively investigated by an operation of UASB reactor (198 days) and a series of batch tests. The results found that PHE was prone to accumulate in reactor by sludge adsorption (Final concentration = 12.53 mg/g TS Sludge), which posed significant influences on methanogenic system. The removal of chemical oxygen demand (COD), NH4+-N and volatile fatty acids (VFAs) in reactor were reduced with PHE accumulation. Meanwhile, microbes with higher ATPase secrete more EPS activity to self-protect against PHE toxicity. Sequencing analysis showed that PHE interfered significantly diversity and structure of microbial community. For bacteria, PHE was toxic to Bacteroidetes and Latescibacteria, while syntrophs (f_Syntrophaceae, Syntrophorhabdus, etc.) involved in VFAs oxidation and aromatic organics degradation were tolerant of PHE stress. For archaea, acetoclastic methanogens (Methanosaeta) abundance was continuously diminished by 45.1% under long-term PHE exposure. Further functions analysis suggested that microbial community accelerated amino acid metabolism, energy metabolism and xenobiotics biodegradation & metabolism to satisfy physiological demanding under PHE stress. Combining batch tests of methanogenic metabolism proved that acetoclastic methanogenesis was negatively affected by PHE due to inhibition of functional enzymes (acetate kinase, phosphate acetyltransferase, etc.) expression. These findings may provide the basis for enhancing bioremediation of PAH pollution in anaerobic environment.

New mechanistic insights into PAHs transport across wheat root cell membrane: Evidence for ABC transporter mediation

Polycyclic aromatic hydrocarbons (PAHs) are a class of highly carcinogenic organic pollutants. Our previous results revealed that the active uptake of PAHs by plant roots is performed through H+/PAHs co-transport. However, the proteins and mechanisms of co-transport of PAHs remain unknown. We hypothesized that ABC transporters are involved in PAHs co-transport via the roots. We found a total of 47 ABC transporters with alkalinity and hydrophobicity which were up-regulated or newly expressed in the wheat roots after phenanthrene exposure. And the concentration of ABC transporters rose. There was a positive relationship between the concentration of phenanthrene and ABC transporter expression in the wheat roots. Additionally, the trend observed in the ABC transporters expression was also found in the gene expression. With energies below −6 kcal mol−1, a stable docking conformation formed between ABC transporters and PAHs. π-π stacking and van der Waals force bound PAHs to ABCB or ABCG. The binding strength of ABCB subfamily proteins with homodimers is stronger than that of ABCG subfamily proteins with single molecules. ABC transporters may transport PAHs by forming a dimer-shaped pocket, translocating it into cells, then opening it within the cells, to release the bound PAHs. These results contributed to our understanding of how ABC transporters aid plant root uptake of PAHs.

Sex-specific thyroid disruption caused by phenanthrene in adult zebrafish (Danio rerio)

Polycyclic aromatic hydrocarbons (PAHs) are well-known contaminants with widespread distribution in environment and food. Phenanthrene is one of the most abundant PAHs in food and aquatic environment and generates reproductive and developmental toxicity in zebrafish. Nonetheless, whether phenanthrene caused sex-specific thyroid disruption in adult zebrafish is unclear. To determine this, adult zebrafish (male and female) were treated with phenanthrene (0, 0.85, 8.5, and 85 μg/L) for 60 days. After the treatment period, we assessed the concentrations of thyroid hormones (THs) and expression levels of genes in the hypothalamic-pituitary-thyroid (HPT) axis. The results showed that phenanthrene exposure can lead to thyroid disruption in both male and female zebrafish. Exposure to phenanthrene dramatically reduced the levels of L-thyroxine (T4) and L-triiodothyronine (T3) in both male and female zebrafish, with a similar trend in both. However, the genes expression profiles of hypothalamic-pituitary-thyroid (HPT) axis were sex-specific. In all, the present study demonstrated that phenanthrene exposure could result in sex-specific thyroid disruption in adult zebrafish.

Exposure to phenanthrene affects oocyte meiosis by inducing mitochondrial dysfunction and endoplasmic reticulum stress.

Phenanthrene (PHE) is one of the most abundant polycyclic aromatic hydrocarbons (PAHs), which is a widespread environmental contaminant. Various studies showed that PHE has adverse impacts on animals and human health. It has been shown that PHE exposure induced follicular atresia and endocrine dyscrasia in female mice. However, the potential mechanism regarding how PHE affects female reproductive system especially the oocyte quality has not been elucidated. In this study, we set up PHE exposure model and found that PHE exposure compromised oocytes maturation competence by inhibiting spindle assembly and chromosomes alignment. Moreover, PHE exposure induced mitochondrial dysfunction and endoplasmic reticulum (ER) stress, leading to increased reactive oxygen species (ROS) and aberrant calcium levels in cytoplasm, eventually induced oxidative stress and DNA damage in oocytes. Furthermore, we found that oral administration of PHE caused the occurrence of oxidative stress and apoptosis in female ovary. In addition, the oocyte exhibited aberrant spindle morphology and failure of actin cap formation in metaphase II oocytes. Taken together, our study demonstrated that mitochondrial dysfunction and ER stress-induced oxidative stress and DNA damage are the major cause of poor oocyte quality after PHE exposure.

Hibiscus sabdariffa Extract Protects HaCaT Cells against Phenanthrene-Induced Toxicity through the Regulation of Constitutive Androstane Receptor/Pregnane X Receptor Pathway.

Phenanthrene (Phe) exposure is associated with skin ageing, cardiotoxicity and developmental defects. Here, we investigated the mode of Phe toxicity in human keratinocytes (HaCaT cells) and the attenuation of toxicity on pre-treatment (6 h) with ethanol extract of Hibiscus sabdariffa calyxes (HS). Cell viability, reactive oxygen species (ROS) generation, mitochondrial membrane potential (ΔΨm) alteration, changes in the transcriptional activity of selected genes involved in phase I and II metabolism, antioxidant response and gluconeogenesis, western blot and docking studies were performed to determine the protective effect of HS against Phe. Phe (250 μM) induced cytotoxicity in HaCaT cells through AhR-independent, CAR/PXR/RXR-mediated activation of CYP1A1 and the subsequent alterations in phase I and II metabolism genes. Further, CYP1A1 activation by Phe induced ROS generation, reduced ΔΨm and modulated antioxidant response, phase II metabolism and gluconeogenesis-related gene expression. However, pre-treatment with HS extract restored the pathological changes observed upon Phe exposure through CYP1A1 inhibition. Docking studies showed the site-specific activation of PXR and CAR by Phe and inhibition of CYP1A1 and CYP3A4 by the bioactive compounds of HS similar to that of the positive controls tested. Our results conclude that HS extract can attenuate Phe-induced toxicity in HaCaT cells through CAR/PXR/RXR mediated inhibition of CYP1A1.

Mechanistic insights into phenanthrene acropetal translocation via wheat xylem: Separation and identification of transfer proteins.

Polycyclic aromatic hydrocarbons (PAHs) have the potential to cause cancer, teratogenicity, and mutagenesis in humans. Long-term plant safe production relies on how PAHs are transported and coordinated across organs. However, the acropetal transfer mechanism of PAHs in staple crop stems, particularly in xylem, a critical path, is unknown. Herein, we first confirmed the presence of specific interaction between the proteins and phenanthrene by employing the magnetic phenanthrene-bound bead immunoassay and label free liquid chromatograph mass spectrometer (LC-MS/MS), suggesting that peroxidase (uniprot accession: A0A3B5XXD0) and unidentified proteins (uniprot accession: A0A3B6LUC6) may function as the carriers to load and acropetally translocate phenanthrene (a model PAH) in wheat xylem. This specified binding of protein-phenanthrene may form through hydrophobic interactions in the conservative binding region, as revealed by protein structural investigations and molecular docking. To further investigate the role of these proteins in phenanthrene solubilization, phenanthrene exposure was conducted: a substantial quantity of peroxidase was produced; an unusually high expression of uncharacterized proteins was observed, indicating their positive effects in the acropetal transfer of phenanthrene in wheat xylem. These data confirmed that the two proteins are crucial in the solubilization of phenanthrene in wheat xylem sap. Our findings provide fresh light on the molecular mechanism of PAH loading in plant xylem and techniques for ensuring the security of staple crops and improving the efficacy of phytoremediation in a PAH-contaminated environment.

AMER3 variants modify the U-shaped association of urinary total hydroxyphenanthrene with fasting plasma glucose: A newfound gene-environment interaction

As a polycyclic aromatic hydrocarbon, environmental exposure to phenanthrene is widespread worldwide. The potential effects and mechanism of phenanthrene exposure on fasting plasma glucose (FPG) have not been well determined. In this study, we aim to explore the effects of phenanthrene exposure and AMER3 variants on fasting plasma glucose (FPG) through a longitudinal epidemiological study. Repeated measurements of five urinary hydroxyphenanthrene (OHPh) for 5739 participants with 7083 observations from the Wuhan-Zhuhai cohort were performed to analyze the relationships between total OHPh (ΣOHPh) and FPG using linear mixed models and restricted cubic spline functions. Then, we genotyped 2777 participants (4104 observations) using the Infinium OmniZhongHua-8 BeadChip and included all 14 single nucleotide polymorphisms (SNPs) within the AMER3 gene to analyze the interaction of the AMER3 on the relationship between ΣOHPh and FPG. We observed a U-shaped relationship between ΣOHPh and FPG, and the turning point of ΣOHPh was 2.512 μg/mmol Cr. When lower than the turning point, ΣOHPh was negatively associated with FPG, while higher than the turning point, ΣOHPh was positively associated with FPG. Furthermore, we observed interactions (Pint <0.05) between two common variants (rs72854995 and rs72854999) of the AMER3 and ΣOHPh on FPG change: the U-shaped relationship was still observed in the GG genotype groups but not in the allele A carriers. Our results suggested that the AMER3 gene can modify the U-shaped relationship between phenanthrenes exposure and FPG, which showed a new gene-environment interaction and will provide a new perspective on the relationship between phenanthrene exposure and FPG.