ABSTRACT
A macrolide antibiotic, lasiodiplodin was isolated from the endophytic fungus (EF) Lasiodiplodia pseudotheobromae J-10 associated with the medicinal plant Sarcandra glabra. In vitro antifungal assay demonstrated the inhibitory activity of lasiodiplodin against the growth of six phytopathogenic fungi, with the IC50 values ranging between 15.50 and 52.30 µg/mL. The highest antifungal activities were recorded against Exserohilum turcicum, Colletotrichum capsici, and Pestalotiopsis theae, with IC50 values of 15.50, 15.90, and 17.55 µg/mL, respectively. The underlying mechanism of the antifungal activity of lasiodiplodin against E. turcicum included the alteration of its colony morphology and disturbance of its cell membrane integrity. In addition, the optimization of L. pseudotheobromae J-10 culture conditions increased lasiodiplodin yield to 52.33 mg/L from 0.59 mg/L at pre-optimization. This is the first report on the isolation and identification of antifungal compound from the EF L. pseudotheobromae J-10 associated with S. glabra, as well as on the optimization of L. pseudotheobromae J-10 culture conditions to increase lasiodiplodin yield. The results of this study support that lasiodiplodin is a natural compound with high potential bioactivity against phytopathogens, and provide a basis for further study of the EF associated with S. glabra.
Subject(s)
Plants, Medicinal , Zearalenone , Antifungal Agents/pharmacology , Zearalenone/pharmacologyABSTRACT
Zearalenone is an estrogenic mycotoxin which is a common food contaminant and has been implicated in increasing the incidence of carcinogenesis and other reproductive health ailments through the estrogen receptor alpha (ERα) pathway. Competitive ERα blockers such as 4-Hydroxytamoxifen (OHT), are synthetic FDA approved drugs which, albeit being an effective anticancer agent, induces life altering side effects. For this reason, there is an increased interest in the use of naturally occurring medicinal plant products such as flavonoids. This study aimed to identity flavonoid ERα inhibitors and provide insights into the mechanism of inhibition using computational techniques. The Molecular Mechanics/Generalized Born Surface Area calculations revealed that quercetrin, hesperidin, epigallocatechin 3-gallate and kaempferol 7-O-glucoside out of 14 flavonoids had higher binding affinity for ERα than OHT. The structural analysis revealed that the binding of the compounds to the receptor lead to dynamic alterations, which induced conformational shift in the structure and orientation of the receptor resulting in stabilised, compact and low energy systems. The results of this study provide imperative information that supports the use of flavonoids in the inhibition of ERα to prevent or ameliorate the consequential adverse effects associated with zearalenone exposure.Communicated by Ramaswamy H. Sarma.
Subject(s)
Receptors, Estrogen , Zearalenone , Receptors, Estrogen/chemistry , Estrogen Receptor alpha , Molecular Dynamics Simulation , Flavonoids/pharmacology , Flavonoids/therapeutic use , Zearalenone/pharmacology , EstrogensABSTRACT
Zearalenone (ZEL)-induced apoptosis in different cells is mediated by various molecular mechanisms, including endoplasmic reticulum (ER) stress. Selenium, an inorganic micronutrient, has several cytoprotective properties, but its potential protective action against ZEL-induced apoptosis in trophoblast cells and the precise mechanisms remain unclear. In this study, we investigated the effects of sodium selenite, a predominant chemical form of selenium, on cell viability, apoptosis, and progesterone (P4) production in ZEL-treated goat trophoblast cell line and explored the underlying molecular mechanisms. ZEL treatment repressed cell viability and promoted apoptosis, which was accompanied by an enhancement of the activity of caspase 3, a key executioner of apoptosis. ZEL treatment was involved in the upregulation of malonaldehyde (MDA) levels and was implicated in the reduction of the protein expression of selenoprotein S (SELS), thereby triggering protein expression of ER stress biomarkers (glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP)). However, sodium selenite attenuates these adverse effects, including increases in apoptotic rate, caspase 3 activity, MDA, GRP78, and CHOP expression and decreases in SELS expression in cells treated with ZEL or Thapsigargin (Tg, an ER stress agonist). Simultaneously, 4-phenylbutyric acid (4-PBA, an ER stress antagonist) treatment significantly alleviated the ZEL-induced deleterious effects on cells in response to ZEL, similarly to sodium selenite. In addition, sodium selenite supplementation effectively rescued the ZEL-induced decrease in P4 production in ZEL-treated cells. In summary, these findings suggest that ZEL triggers apoptosis in goat trophoblast cells by downregulating SELS expression and activating the ER stress signaling pathway and that sodium selenite protects against these detrimental effects. This study provides novel insights into the benefits of using selenium against ZEL-induced apoptosis and cellular damage.
Subject(s)
Selenium , Zearalenone , Animals , Apoptosis , Caspase 3 , Endoplasmic Reticulum Stress/physiology , Goats/metabolism , Selenium/pharmacology , Sodium Selenite/pharmacology , Transcription Factor CHOP/genetics , Transcription Factor CHOP/metabolism , Transcription Factor CHOP/pharmacology , Trophoblasts/metabolism , Zearalenone/pharmacologyABSTRACT
Fridericia chica (Bignoniaceae) is a traditional medicinal plant. The aim of this research was to determine the protective effects of the hydroethanolic extract from the F. chica leaves (HEFc) against the cytotoxicity of zearalenone (α-ZEL) and ß-ZEL on SH-SY5Y cells. Free radical scavenging activity of HEFc was evaluated using the DPPH method. The cytotoxicity of both zearalenone metabolites and HEFc was examined using MTT test, as was the cytoprotective effects of the HEFc on cells treated with these mycotoxins. The chemical composition of HEFc was determined using UPLC-QTOF-MS/MS. HEFc elicited good DPPH radical scavenging activity following a concentration-dependent relationship. Cells exposed to α-ZEL exhibited a viability Ë50% after 48 h of treatment (25 and 50 µM), while those exposed to ß-ZEL showed viability Ë50% (100 µM) and Ë25% (25-100 µM) after 24 and 48 h of exposure, respectively. HEFc showed a significant increase in cell viability after exposure to α-ZEL (25 and 50 µM) and ß-ZEL (6-100 µM) (p < 0.05). UPLC-QTOF-MS/MS analyses allowed the identification of 10 phytochemical components in the HEFc. In short, the hydroethanolic extract of F. chica grown in Colombian Caribbean can protect against the effects of mycotoxins and it is a valuable source of compounds with antioxidant properties.
Subject(s)
Bignoniaceae/chemistry , Neuroblastoma/metabolism , Plant Extracts/pharmacology , Protective Agents/pharmacology , Zearalenone/pharmacology , Cell Line, Tumor , Humans , Plant Extracts/chemistry , Plant Leaves/chemistry , Protective Agents/chemistryABSTRACT
INTRODUCTION: Endophyte is considered a source of natural bioactive secondary metabolites that provides an array of bioactive lead compounds. The present study was aimed to determine the antimicrobial and anti-inflammatory potential of fungal endophytes isolated from Catharanthus roseus. METHODS: A total of seven fungal endophytes crude extract were screened against bacterial pathogens. Of these, Curvularia geniculata CATDLF7 crude extract exhibited the most potent inhibitory activity against bacterial pathogens. Hence, CATDLF7 crude extract was subjected to chromatographic separation. This purification leads to the isolation of six pure compounds (1PS - 6PS). Of these, 3PS was found to be a major constituent and most effective against clinical isolates of methicillin- resistant Staphylococcus aureus (MRSA) with minimum inhibitory concentration (MIC) values ranging from 100 to 200 µg/ml. Based on the spectroscopic data, 3PS was characterized as α,ß- dehydrocurvularin. This compound also showed synergistic interaction with norfloxacin and reduced its MIC up to 32-folds with a fractional inhibitory concentration index (FICI) of 0.09. RESULTS: To understand the possible antibacterial mechanism of action, α,ß-dehydrocurvularin alone (100 µg/ml) exhibited efflux pump inhibitory potential by 0.84 fold decreasing in ethidium bromide (EtBr) fluorescence. In addition, α,ß-dehydrocurvularin inhibited inflammatory cytokines TNF-α and IL-6 production, which is further validated by molecular docking scores -4.921 and -5.641, respectively, for understanding orientation and binding affinity. CONCLUSION: Overall, the results highlighted identifying bioactive compound α,ß-dehydrocurvularin, which could be used as an antimicrobial and anti-inflammatory agent.
Subject(s)
Anti-Infective Agents/isolation & purification , Anti-Inflammatory Agents/isolation & purification , Catharanthus/chemistry , Plant Extracts/isolation & purification , Plant Leaves/chemistry , Zearalenone/analogs & derivatives , Animals , Anti-Infective Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Cytokines/metabolism , Disease Models, Animal , Drug Evaluation, Preclinical , Drug Therapy, Combination , Endophytes/metabolism , Female , Humans , Interleukin-6/metabolism , Methicillin-Resistant Staphylococcus aureus/drug effects , Mice , Microbial Sensitivity Tests , Molecular Docking Simulation , Norfloxacin/pharmacology , Plant Extracts/pharmacology , Protein Binding , Signal Transduction , Structure-Activity Relationship , Zearalenone/isolation & purification , Zearalenone/pharmacologyABSTRACT
Recently, the palbociclib/letrozole combination therapy was granted accelerated US FDA approval for the treatment of estrogen receptor (ER)-positive breast cancer. Since the underlying metabolic effects of these drugs are yet unknown, we investigated their synergism at the metabolome level in MCF-7 cells. As xenoestrogens interact with the ER, we additionally aimed at deciphering the impact of the phytoestrogen genistein and the estrogenic mycotoxin zearalenone. A global metabolomics approach was applied to unravel metabolite and pathway modifications. The results clearly showed that the combined effects of palbociclib and letrozole on cellular metabolism were far more pronounced than that of each agent alone and potently influenced by xenoestrogens. This behavior was confirmed in proliferation experiments and functional assays. Specifically, amino acids and central carbon metabolites were attenuated, while higher abundances were observed for fatty acids and most nucleic acid-related metabolites. Interestingly, exposure to model xenoestrogens appeared to counteract these effects.
Subject(s)
Letrozole/pharmacology , Metabolome/drug effects , Phytoestrogens/pharmacology , Piperazines/pharmacology , Pyridines/pharmacology , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Carbon/metabolism , Diet , Female , Genistein/chemistry , Genistein/pharmacology , Humans , Letrozole/chemistry , Letrozole/therapeutic use , MCF-7 Cells , Metabolomics , Phytoestrogens/chemistry , Piperazines/chemistry , Piperazines/therapeutic use , Principal Component Analysis , Pyridines/chemistry , Pyridines/therapeutic use , Receptors, Estrogen/metabolism , Zearalenone/chemistry , Zearalenone/pharmacologyABSTRACT
It is now established that mycoestrogen zearalenone (ZEN) disrupts reproductive physiology, but the specific mechanisms by which this occurs remain unknown, especially in brain. Growing evidence suggests that populations of estradiol (E2)-sensitive neurons in anteroventral periventricular (AVPV) and arcuate (ARC) nuclei, especially kisspeptin neurons, play a pivotal role in the timing of puberty onset, ovulation, and normal reproduction. The present study was conducted to find whether the ZEN can cause estrogen-like actions during the critical period of neonatal differentiation. In this study, we compared the effect of neonatal exposure to sesame oil, E2 benzoate (EB, 20 µg/kg body weight [bw]), and 3 various doses: 0.2, 1, and 2 mg/kg bw of ZEN (0.2, 1, and 2 ZEN) on the onset of puberty and estrus cyclicity as well as ovarian follicular profile, kisspeptin expression, and neuronal density in AVPV and ARC hypothalamic nuclei and E2 and luteinizing hormone (LH) levels on postnatal day 70. Control mice received no treatment. Vaginal opening was significantly advanced by EB and 2 ZEN. Disrupted estrus cycles and decreased follicular profiles were observed in EB, 1 ZEN, and 2 ZEN animals. In addition, EB, 1 ZEN, and 2 ZEN reduced the expression of kisspeptin and neuronal density of AVPV and ARC nuclei and caused a decrease in the LH and an increase in E2 plasma levels. Taken together, our observations provide physiological evidence that neonatal exposure to ZEN exerts estrogen-like actions in the estrogen-sensitive hypothalamic AVPV and ARC nuclei, controlling reproductive functions in adult female mice.
Subject(s)
Estrous Cycle/drug effects , Hypothalamus/drug effects , Reproduction/drug effects , Sexual Maturation/drug effects , Zearalenone/pharmacology , Animals , Dose-Response Relationship, Drug , Estradiol/analogs & derivatives , Estradiol/blood , Estradiol/pharmacology , Estrous Cycle/metabolism , Female , Hypothalamus/metabolism , Kisspeptins/metabolism , Luteinizing Hormone/blood , Mice , Mice, Inbred BALB C , Neurons/drug effects , Neurons/metabolism , Ovarian Follicle/drug effects , Ovarian Follicle/metabolismABSTRACT
Estrogen receptors (ERs) α and ß are distributed in most tissues of women and men. ERs are bound by estradiol (E2), a natural hormone, and mediate the pleiotropic and tissue-specific effects of E2, such as proliferation of breast epithelial cells or protection and differentiation of neuronal cells. Numerous environmental molecules, called endocrine disrupting compounds, also interact with ERs. Phytoestrogens belong to this large family and are considered potent therapeutic molecules that act through their selective estrogen receptor modulator (SERM) activity. Using breast cancer cell lines as a model of estrogen-dependent proliferation and a stably ER-expressing PC12 cell line as a model of neuronal differentiating cells, we studied the SERM activity of major dietary compounds, such as apigenin, liquiritigenin, daidzein, genistein, coumestrol, resveratrol and zearalenone. The ability of these compounds to induce ER-transactivation and breast cancer cell proliferation and enhance Nerve Growth Factor (NGF) -induced neuritogenesis was assessed. Surprisingly, although all compounds were able to activate the ER through an estrogen responsive element reporter gene, they showed differential activity toward proliferation or differentiation. Apigenin and resveratrol showed a partial or no proliferative effect on breast cancer cells but fully contributed to the neuritogenesis effect of NGF. However, daidzein and zearalenone showed full effects on cellular proliferation but did not induce cellular differentiation. In summary, our results suggest that the therapeutic potential of phytoestrogens can diverge depending on the molecule and the phenotype considered. Hence, apigenin and resveratrol might be used in the development of therapeutics for breast cancer and brain diseases.
Subject(s)
Adrenal Gland Neoplasms/drug therapy , Antineoplastic Agents, Phytogenic/pharmacology , Breast Neoplasms/drug therapy , Cell Proliferation/drug effects , Diet , Neurogenesis/drug effects , Pheochromocytoma/drug therapy , Phytoestrogens/pharmacology , Selective Estrogen Receptor Modulators/pharmacology , Adrenal Gland Neoplasms/genetics , Adrenal Gland Neoplasms/metabolism , Adrenal Gland Neoplasms/pathology , Animals , Apigenin/pharmacology , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Chemokine CXCL12/genetics , Chemokine CXCL12/metabolism , Dose-Response Relationship, Drug , Estrogen Receptor alpha/drug effects , Estrogen Receptor alpha/genetics , Estrogen Receptor alpha/metabolism , Female , Gene Expression Regulation, Neoplastic/drug effects , Humans , Isoflavones/pharmacology , MCF-7 Cells , Nerve Tissue Proteins/metabolism , Neurites/drug effects , Neurites/metabolism , Neurites/pathology , PC12 Cells , Pheochromocytoma/genetics , Pheochromocytoma/metabolism , Pheochromocytoma/pathology , Rats , Response Elements , Resveratrol , Stilbenes/pharmacology , Transcription, Genetic/drug effects , Transfection , Zearalenone/pharmacologyABSTRACT
Fungal endophytes offer diverse and unique secondary metabolites, making these organisms potential sources of promising drug leads. The application of high-resolution-liquid chromatography mass spectrometry and nuclear magnetic resonance-based metabolomics to fungal endophytes is practical in terms of dereplication studies and the mining of bioactive compounds. In this paper, we report the application of metabolomics in parallel with anti-trypanosomal assays to determine the ideal conditions for the medium-scale fermentation of the endophyte Lasiodiplodia theobromae. The 1H NMR comparison between the active versus inactive fractions identified several unique chemical fingerprints belonging to the active fractions. Furthermore, by integrating high-resolution-liquid chromatography mass spectrometry data with multivariate data analysis, such as orthogonal partial least squares-discriminant analysis (OPLS-DA) and the bioactivity results of the fractions of L. theobromae, the anti-trypanosomal agents were easily discerned. With available databases such as Antibase and Dictionary of Natural Products coupled to MZmine through in-house algorithms optimized in our laboratory, the predicted metabolites were readily identified prior to isolation. Fractionation was performed on the active fractions and three known compounds were isolated, namely, cladospirone B, desmethyl-lasiodiplodin, and R-(-)-mellein. Cladospirone B and desmethyl-lasiodiplodin were among the predicted compounds generated by the OPLS-DA S-plot, and these compounds exhibited good activity against Trypanosoma brucei brucei with minimum inhibitory concentrations of 17.8 µM and 22.5 µM, respectively.
Subject(s)
Ascomycota/chemistry , Dioxins/isolation & purification , Trypanocidal Agents/isolation & purification , Trypanosoma brucei brucei/drug effects , Zearalenone/analogs & derivatives , Algorithms , Ascomycota/metabolism , Biological Products , Chromatography, Liquid , Dioxins/chemistry , Dioxins/pharmacology , Endophytes/chemistry , Endophytes/metabolism , Magnetic Resonance Spectroscopy , Mass Spectrometry , Metabolomics/methods , Microbial Sensitivity Tests , Trypanocidal Agents/chemistry , Trypanocidal Agents/pharmacology , Zearalenone/chemistry , Zearalenone/isolation & purification , Zearalenone/pharmacologyABSTRACT
KRAS mutant lung cancers have long been considered as untreatable with drugs. Transforming growth factor-ß-activated kinase 1 (TAK1) appears to play an anti-apoptotic role in response to multiple stresses and has been reported to be a responsive kinase that regulates cell survival in KRAS-dependent cells. In this study, in order to find a useful approach to treat KRAS mutant lung cancer, we focused on the combined effects of 5Z-7-oxozeaenol, a TAK1 inhibitor, with hyperthermia (HT) in KRAS mutant lung cancer cell line A549. Annexin V-FITC/PI assay, cell cycle analysis, and colony formation assay revealed a significant enhancement in apoptosis induced by HT treatment, when the cells were pre-incubated with 5Z-7-oxozeaenol in a dose-dependent manner. The enhanced apoptosis by 5Z-7-oxozeaenol was accompanied by a significant increase in reactive oxygen species (ROS) generation and loss of mitochondrial membrane potential (MMP). In addition, western blot showed that 5Z-7-oxozeaenol enhanced HT-induced expressions of cleaved caspase-3, cleaved caspase-8, and HSP70 and decreased HT-induced expressions of Bcl-2, p-p38, p-JNK, and LC3. Moreover, 5Z-7-oxozeaenol pre-treatment resulted in a marked elevation of intracellular calcium level which might be associated with endoplasmic reticulum (ER) stress-related pathway. Taken together, our data provides further insights of the mechanism of action of 5Z-7-oxozeaenol and HT treatment, and their potential application as a novel approache to treat patients with KRAS mutant lung cancer.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis , Lung Neoplasms/therapy , Zearalenone/analogs & derivatives , A549 Cells , Combined Modality Therapy , Drug Screening Assays, Antitumor , Endoplasmic Reticulum Stress , Humans , Hyperthermia, Induced , MAP Kinase Kinase Kinases/antagonists & inhibitors , Membrane Potential, Mitochondrial/drug effects , Reactive Oxygen Species/metabolism , Zearalenone/pharmacologyABSTRACT
Healthy male Kunming mice received selenium yeast for 14 days prior to a single oral administration of zearalenone (ZEN). After 48 h, blood samples were collected for analysis and showed that mice in the ZEN-treated group has significantly decreased lymphocytes (P < 0.05) and platelets (P < 0.05) along with an increased white blood cell (WBC) count and other constituents (P < 0.05). The serum biochemistry analysis of the ZEN group indicated that glutamic pyruvic transaminase (ALT), glutamic oxaloacetic transaminase (AST), urea, and uric acid were significantly increased (P < 0.05), whilst total bilirubin (TB) and albumin (ALB) were decreased along with serum testosterone and estrogen (P < 0. 05). The level of malondialdehyde (MDA) in the serum of the ZEN group was significantly increased whilst glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) had significantly decreased (P < 0.05). Treatment with selenium yeast had a significant effect on response with most of the experimental parameters returning to levels similar to those observed in the untreated control mice. From these data, it can be concluded that ZEN is highly poisonous in Kunming mice with high levels of toxicity on the blood, liver, and kidneys. High levels of oxidative stress were observed in mice and pre-treatment with selenium yeast by oral gavage is effective in the ameliorated effects of ZEN-induced damage.
Subject(s)
Gonadal Steroid Hormones/blood , Oxidative Stress/drug effects , Saccharomyces cerevisiae , Selenium/pharmacology , Zearalenone/adverse effects , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Bilirubin/blood , Male , Malondialdehyde/blood , Mice , Urea/blood , Uric Acid/blood , Zearalenone/pharmacology , Zearalenone/toxicityABSTRACT
The ATPase p97 is a ubiquitin targeted segregase that uses the energy of ATP binding and hydrolysis to extract ubiquitylated substrates from biological membranes, from other proteins, or from protein complexes to carry out myriad tasks in eukaryotes. Increased p97 activity has been linked to a poor prognosis in cancer patients, making p97 an anti-neoplastic target. In the present study, we show that dehydrocurvularin (DHC) and its chlorinated variants are covalent inhibitors of p97, interfering with its ATPase activity. Interestingly, cellular studies revealed both DHC and its monochloro analogue interfere with both the proteasome and p97, whereas its dichloro analogue showed p97 specificity.
Subject(s)
Adenosine Triphosphatases/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Halogenation , Nuclear Proteins/antagonists & inhibitors , Zearalenone/analogs & derivatives , Adenosine Triphosphatases/metabolism , Drug Evaluation, Preclinical , Enzyme Inhibitors/metabolism , Nuclear Proteins/metabolism , Substrate Specificity , Zearalenone/chemistry , Zearalenone/metabolism , Zearalenone/pharmacologyABSTRACT
Xenoestrogens from synthetic or natural origin represent an increasing risk of disrupted endocrine functions including the physiological activity of the hypothalamo-pituitary-gonad axis. Ethinyl estradiol (EE2) is a synthetic estrogen used in contraceptive pills, whereas zearalenone (ZEA) is a natural mycoestrogen found with increasing prevalence in various cereal crops. Both EE2 and ZEA are agonists of estrogen receptor-α and accelerate puberty. However, the neuroendocrine mechanisms that are responsible for this effect remain unknown. Immature female Wistar rats were treated with EE2 (10 µg/kg), ZEA (10 mg/kg), or vehicle for 10 days starting from postnatal day 18. As a marker of puberty, the vaginal opening was recorded and neuropeptide and related transcription factor mRNA levels were measured by quantitative real time PCR and in situ hybridization histochemistry. Both ZEA and EE2 accelerated the vaginal opening, increased the uterine weight and the number of antral follicles in the ovary, and resulted in the increased central expression of gnrh. These changes occurred in parallel with an earlier increase of kiss1 mRNA in the anteroventral and rostral periventricular hypothalamus and an increased kisspeptin (KP) fiber density and KP-GnRH appositions in the preoptic area. These changes are compatible with a mechanism in which xenoestrogens overstimulate the developmentally unprepared reproductive system, which results in an advanced vaginal opening and an enlargement of the uterus at the periphery. Within the hypothalamus, ZEA and EE2 directly activate anteroventral and periventricular KP neurons to stimulate GnRH mRNA. However, GnRH and gonadotropin release and ovulation are disrupted due to xenoestrogen-mediated inhibitory KP signaling in the arcuate nucleus.
Subject(s)
Ethinyl Estradiol/pharmacology , Kisspeptins/metabolism , Sexual Maturation/drug effects , Zearalenone/pharmacology , Animals , Arcuate Nucleus of Hypothalamus/drug effects , Arcuate Nucleus of Hypothalamus/metabolism , Estrogens/pharmacology , Estrogens, Non-Steroidal/pharmacology , Female , Gene Expression/drug effects , Gonadotropin-Releasing Hormone/genetics , Gonadotropin-Releasing Hormone/metabolism , Gonadotropins/metabolism , Hypothalamus/cytology , Hypothalamus/drug effects , Hypothalamus/metabolism , In Situ Hybridization , Kisspeptins/genetics , Microscopy, Confocal , Neurons/drug effects , Neurons/metabolism , Rats, Wistar , Receptors, G-Protein-Coupled/genetics , Receptors, G-Protein-Coupled/metabolism , Receptors, Kisspeptin-1 , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effects , Signal Transduction/genetics , Uterus/drug effects , Uterus/growth & development , Uterus/metabolism , Xenobiotics/pharmacologyABSTRACT
Zearalenone (ZEA) is a mycotoxin produced by various Fusarium fungi, which has been shown to cause several cases of mycotoxicosis in farm animals and humans. However, there is no evidence regarding the effect of ZEA on mouse egg developmental competence. In this study, we found that the activation rate of maturated oocytes was affected in mice by ZEA treatment, indicating that ZEA affects egg developmental competence. And we explored possible mechanisms of low mouse maturated oocyte developmental competence after ZEA treatment from an epigenetic modification perspective. The fluorescence intensity analysis showed that 5-methyl cytosine level increased after ZEA treatment, indicating that the general DNA methylation level increased in the treated eggs. Moreover, histone methylations were also altered: H3K4me2 as well as H3K9me3 and H4K20me1, me2, me3 levels decreased in eggs that were cultured in high-dose ZEA medium. Thus, our results indicated that ZEA decreased egg developmental competence by affecting the epigenetic modifications.
Subject(s)
Epigenesis, Genetic/drug effects , Ovum/metabolism , Zearalenone/pharmacology , Animals , Cell Differentiation/drug effects , DNA Methylation/drug effects , Female , Histones/metabolism , Humans , Lysine/metabolism , Mice, Inbred ICR , Ovum/drug effectsABSTRACT
Large doses of bone morphogenetic protein 2 (BMP2) are used clinically to induce bone formation in challenging bone defects. However, complications after treatment include swelling, ectopic bone formation, and adjacent bone resorption. While BMP2 can be effective, it is important to characterize the mechanism of the deleterious effects to optimize its use. The aim of this study was to determine the effect of BMP2 on apoptosis in osteoblast lineage cells and to determine the role of the BMP inhibitor Noggin in this process. Human mesenchymal stem cells (MSCs), immature osteoblast-like MG63 cells, and mature normal human osteoblasts (NHOst) were treated with BMP2. A model system of increased endogenous BMP signaling was created by silencing Noggin (shNOG-MG63). Finally, the BMP pathway regulating apoptosis in NHOst was examined using BMP signaling inhibitors (5Z-7-oxozeaenol, dorsomorphin, H-8). Apoptosis was characterized by caspase-3, BAX/BCL2, p53, and DNA fragmentation. BMP2 induced apoptosis in a cell-type dependent manner. While the effect was minor in MSCs, MG63 cells had modest increases and NHOst cells had robust increases apoptosis after BMP2 treatment. Apoptosis was significantly higher in shNOG-MG63 than MG63 cells. 5Z-7-oxozeaenol and dorsomorphin eliminated the BMP2-induced increase in DNA fragmentation in NHOst, suggesting roles for TAB/TAK1 and Smad signaling. These results indicate that the apoptotic effect of BMP2 is dependent on cell maturation state, inducing apoptosis in committed osteoblasts through Smad and TAB/TAK1 signaling, and is regulated by Noggin. Dose and delivery must be optimized in therapeutic applications of BMP2 to minimize complications.
Subject(s)
Apoptosis , Bone Morphogenetic Protein 2/pharmacology , Carrier Proteins/metabolism , Osteoblasts/drug effects , Bone Morphogenetic Proteins/antagonists & inhibitors , Carrier Proteins/genetics , Caspase 3/genetics , Caspase 3/metabolism , DNA Fragmentation , Enzyme Activation , Humans , In Situ Nick-End Labeling , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/metabolism , Osteoblasts/metabolism , Pyrazoles/pharmacology , Pyrimidines/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Recombinant Proteins/pharmacology , Signal Transduction , Zearalenone/analogs & derivatives , Zearalenone/pharmacology , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolismABSTRACT
Radish (Raphanus sativus) is a cruciferous plant, rich on flavonoids, isothiocyanates, and phenolic acids. They show anti-inflammatory and immunomodulatory activity both in vitro and in vivo. Isothiocyanates and flavonoids have been reported previously to prevent low-sub-chronic dose of zearalenone (ZEN) causing immunotoxicity. The present study focuses on the amelioration of fusarotoxicosis in Balb/c mice by feeding two concentrations of radish extract. The extract at 15 and 30 mg/kg bw, was evaluated to reduce the deleterious effects in immunological parameters of high subchronic doses of 40 and 80 mg of ZEN/kg bw on modulation of lipopolysaccharide (LPS). ZEN consuming mice showed a "dose-related" decrease in weight gain and in the immune relative weights organs. Moreover, Atrophy and lymphoid depletion were seen in the histopathology of spleen. Ingestion of ZEN at either level had a significant effect on total red blood cell numbers and on their relative number of lymphocytes. Likewise, ZEN alters the production of regulatory cytokines and antibody of LPS stimulated mice. By contrast, the additions of radish extract with a low or high dose of ZEN moderately decreased the affected mice and/or the severity of lesions, and all tested parameters were normal or at least near normal levels. In addition, the radish extract alone did not produce any significant changes in all tested parameters compared with the controls. In conclusion, radish extract was effective for the protection of high dose ZEN-immunotoxication in mice and it could contribute to a solution of the ZEN immunotoxicity in humans and in farm animals.
Subject(s)
Drug-Related Side Effects and Adverse Reactions/prevention & control , Plant Extracts/pharmacology , Raphanus/chemistry , Zearalenone/pharmacology , Zearalenone/toxicity , Administration, Oral , Animal Structures/drug effects , Animal Structures/pathology , Animals , Antibodies/blood , Antibodies/immunology , Antibody Formation/drug effects , Antibody Formation/immunology , Blood/drug effects , Body Weight/drug effects , Drinking/drug effects , Drug-Related Side Effects and Adverse Reactions/immunology , Drug-Related Side Effects and Adverse Reactions/pathology , Eating/drug effects , Erythrocyte Count , Female , Interleukin-1beta/blood , Leukocyte Count , Lipopolysaccharides/immunology , Mice , Mice, Inbred BALB C , Mortality , Organ Size/drug effects , Plant Extracts/administration & dosage , Plant Extracts/therapeutic use , Spleen/drug effects , Spleen/pathology , Tumor Necrosis Factor-alpha/blood , Vaccination , Zearalenone/administration & dosageABSTRACT
Several anthropogenous and naturally occurring substances, referred to as estrogen active compounds (EACs), are able to interfere with hormone and in particular estrogen receptor signaling. EACs can either cause adverse health effects in humans and wildlife populations or have beneficial effects on estrogen-dependent diseases. The aim of this study was to examine global gene expression profiles in estrogen receptor (ER)-proficient Ishikawa plus and ER-deficient Ishikawa minus endometrial cancer cells treated with selected well-known EACs (diethylstilbestrol, genistein, zearalenone, resveratrol, bisphenol A and o,p'-DDT). We also investigated the effect of the pure antiestrogen ICI 182,780 (ICI) on the expression patterns caused by these compounds. Transcript levels were quantified 24 h after compound treatment using Illumina BeadChip Arrays. We identified 87 genes with similar expression changes in response to all EAC treatments in Ishikawa plus. ICI lowered the magnitude or reversed the expression of these genes, indicating ER dependent regulation. Apart from estrogenic gene regulation, bisphenol A, o,p'-DDT, zearalenone, genistein and resveratrol displayed similarities to ICI in their expression patterns, suggesting mixed estrogenic/antiestrogenic properties. In particular, the predominant antiestrogenic expression response of resveratrol could be clearly distinguished from the other test compounds, indicating a distinct mechanism of action. Divergent gene expression patterns of the phytoestrogens, as well as weaker estrogenic gene expression regulation determined for the anthropogenous chemicals bisphenol A and o,p'-DDT, warrants a careful assessment of potential detrimental and/or beneficial effects of EACs. The characteristic expression fingerprints and the identified subset of putative marker genes can be used for screening chemicals with an unknown mode of action and for predicting their potential to exert endocrine disrupting effects.
Subject(s)
Endocrine Disruptors/pharmacology , Endometrial Neoplasms/genetics , Estrogen Antagonists/pharmacology , Estrogens/pharmacology , Gene Expression Profiling , Gene Expression Regulation, Neoplastic/drug effects , Receptors, Estrogen/drug effects , Benzhydryl Compounds , Cell Line, Tumor , Cluster Analysis , DDT/pharmacology , Diethylstilbestrol/pharmacology , Endocrine Disruptors/toxicity , Endometrial Neoplasms/metabolism , Estradiol/analogs & derivatives , Estradiol/pharmacology , Estrogens/toxicity , Female , Fulvestrant , Gene Expression Profiling/methods , Genistein/pharmacology , Humans , Oligonucleotide Array Sequence Analysis , Phenols/pharmacology , Phytoestrogens/pharmacology , Polymerase Chain Reaction , RNA, Messenger/metabolism , Receptors, Estrogen/genetics , Receptors, Estrogen/metabolism , Reproducibility of Results , Resveratrol , Risk Assessment , Stilbenes/pharmacology , Time Factors , Zearalenone/pharmacologyABSTRACT
There is considerable evidence that exogenous estrogenic compounds can have adverse effects on fertility. The main reason cited in literature for hyperestrogenism in pigs is contamination of feedstuffs by the mycotoxin zearalenone (Boehm, 2000), but further estrogenically active substances might also be involved in cases of impaired fertility with symptoms like enlarged, red-coloured vulvae in piglets, irregular estrus cycles and anestrus of sows (Bennetts et al., 1946; Drane et al., 1981). It is well known that soy used in diets for pigs as a main protein source contains phytoestrogens. Amongst them, isoflavones like genistein and daidzein are of particular interest. Aim of this study was to optimize and use an established bioassay (Kluczka, 2003) to determine estrogenic activity in feedstuffs for pigs related to isoflavones and further substances with estrogenic potential. This bioassay is a reporter gene assay based on stably transfected human embryonal kidney cells (HEK 293) that contains either alpha or beta estrogen receptor (alpha- or beta-HEK). The estrogenic activity measured in the luciferase assay was expressed in estradiol-equivalents (EEQ) and the results were compared with the isoflavone content (genistein, daidzein) obtained by chemical analysis using high performance liquid chromatography-Ultraviolet (HPLC-UV). Mean estrogenic activity in diets fed to sows in herds with altered fertility was 275.8 microg EEQ/kg feed in alpha-HEK cells and 295.0 microg EEQ/kg feed in beta-HEK cells. Feedstuffs from herds without any altered fertility showed an average estrogenic activity of 204.9 microg EEQ/kg feed in alpha-HEK and 213.3 microg EEQ/kg feed in beta-HEK. The estrogenic activity was strongly related to the concentration of the isoflavones (alpha-HEK, r(2)=0.9488; beta-HEK, r(2)=0.9427). Clinically relevant zearalenone concentrations (>50-150 microg/kg feed) displayed estrogenic effects in the bioassay that did not differ significantly from those caused by high isoflavone concentration because of the use of soy as protein source.
Subject(s)
Animal Feed/analysis , Biological Assay/veterinary , Chromatography, High Pressure Liquid/veterinary , Phytoestrogens/analysis , Receptors, Estrogen/metabolism , Swine , Animal Nutritional Physiological Phenomena , Animals , Biological Assay/methods , Chromatography, High Pressure Liquid/methods , Estrogen Receptor alpha/drug effects , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta/drug effects , Estrogen Receptor beta/metabolism , Female , Fertility/drug effects , Fertility/physiology , Food Contamination , Genistein/analysis , Genistein/pharmacology , Humans , Isoflavones/analysis , Isoflavones/pharmacology , Phytoestrogens/pharmacology , Receptors, Estrogen/drug effects , Transfection , Tumor Cells, Cultured , Zearalenone/analysis , Zearalenone/pharmacologyABSTRACT
(S)-Curvularin and its 13-, 14-, and 16-membered lactone homologues were synthesized through a uniform strategy in which a Kochi oxidative decarboxylation and ring-closing metathesis reactions constitute the key processes. In the evaluation of the anti-inflammatory effects of the synthesized compounds in assays using cells stably transfected with a human iNOS promoter-luciferase reporter gene construct, the 14- and 16-membered homologues showed a slightly higher inhibitory effect towards iNOS promoter activity than curvularin itself. However, the larger ring homologues also exhibited higher cytotoxicity, manifest in downregulated eNOS promoter activity. In contrast, the di-O-acetyl and 4-chloro derivatives of (S)-curvularin showed higher inhibitory efficiency towards induction of the iNOS promoter and less negative effect on eNOS promoter activity than curvularin.
Subject(s)
Enzyme Inhibitors/chemical synthesis , Gene Expression Regulation, Enzymologic/drug effects , Lactones/chemical synthesis , Nitric Oxide Synthase Type II/antagonists & inhibitors , Zearalenone/analogs & derivatives , Cell Line , Crystallography, X-Ray , Cyclization , Drug Evaluation, Preclinical , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Gene Expression Regulation, Enzymologic/genetics , HeLa Cells , Humans , Lactones/chemistry , Lactones/pharmacology , Models, Molecular , Molecular Structure , Nitric Oxide Synthase Type II/genetics , Promoter Regions, Genetic/drug effects , Promoter Regions, Genetic/genetics , Stereoisomerism , Zearalenone/chemical synthesis , Zearalenone/chemistry , Zearalenone/pharmacologyABSTRACT
Phytoestrogens and mycoestrogens are naturally occurring plant and fungus secondary metabolites with estrogen-like structure and/or actions. We aimed to check the hypothesis that phytoestrogens and mycoestrogens, due to their ability to elicit cerebral vasodilation, can induce acute increases in brain blood perfusion. For this purpose, we continuously recorded cerebrocortical perfusion by laser-Doppler flowmetry in anesthetized rats receiving intracarotid infusions (1 mg/kg) of one of the following estrogenic compounds: biochanin A, daidzein, genistein or zearalanone. We have shown the ability of two isoflavone class phytoestrogens (daidzein and biochanin A) and the mycoestrogen zearalanone to induce acute increases in brain blood flow when locally infused into the cerebral circulation of anesthetized rats. The isoflavone genistein failed to induce a significant increase in brain perfusion. No concomitant changes in blood pressure were recorded during the cerebral effects of the estrogenic compounds. Therefore, these microcirculatory effects were due to direct actions of the estrogenic compounds on the cerebrovascular bed.