RESUMO
Background and Objectives: Polycystic ovary syndrome (PCOS) is one of the most prevalent disorders among women of reproductive age. It is considered as a pro-inflammatory state with chronic low-grade inflammation, one of the key factors contributing to the pathogenesis of this disorder. Polycystic ovary is a well-established criterion for PCOS. The present investigation aimed at finding the role of hyperandrogenism, the most important feature of PCOS, in the development of this inflammatory state. To address this problem, we adopted a model system that developed polycystic ovary morphology (PCOM), which could be most effectively used in order to study the role of non-aromatizable androgen in inflammation in PCOS. Materials and Methods: Six rats were used to induce PCOM in 21-days-old female Wistar albino rats by using a pre-determined release of dihydrotestosterone (DHT), a potent non-aromatizable androgen, achieved by implanting a DHT osmotic pump, which is designed to release a daily dose of 83 µg. Results: After 90 days, the rats displayed irregular estrous cycles and multiple ovarian cysts similar to human PCOS. Elevated serum inflammatory markers such as tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß), and the presence of a necrotic lesion in the liver, osteoclast in the femur, multinucleated giant cells and lymphocytes in the ovary based on histopathological observation of DHT-treated rats clearly indicated the onset of inflammation in the hyperandrogenic state. Our results show no significant alterations in serum hormones such as luteinizing hormone (LH), follicle stimulating hormone (FSH), insulin, and cortisol between control and hyperandrogenised rats. DHT was significantly elevated as compared to control. mRNA studies showed an increased expression level of TNF-α and IL-1ß, further, the mRNA expression of urocortin 1 (Ucn-1) was stupendously elevated in the liver of hyperandrogenised rats. Conclusions: Thus, results from this study provide: (1) a good PCOM model system in order to study the inflammatory changes in PCOS aspects, (2) alteration of inflammatory markers in PCOM rats that could be either due to its direct effect or by the regulation of various inflammatory genes and markers in the liver of hyperandrogenic state suggesting the regulatory role of DHT, and (3) alteration in stress-related protein in the liver of PCOM rats.
Assuntos
Citocinas/sangue , Di-Hidrotestosterona/efeitos adversos , Hiperandrogenismo/metabolismo , Mediadores da Inflamação/sangue , Síndrome do Ovário Policístico/metabolismo , Animais , Modelos Animais de Doenças , Feminino , Hormônio Foliculoestimulante/sangue , Hiperandrogenismo/induzido quimicamente , Insulina/sangue , Fígado/metabolismo , Hormônio Luteinizante/sangue , Síndrome do Ovário Policístico/complicações , Ratos , Ratos Wistar , Urocortinas/metabolismoRESUMO
Endometriosis is a debilitating, estrogen-dependent, progesterone-resistant, inflammatory gynecological disease of reproductive age women. Two major clinical symptoms of endometriosis are chronic intolerable pelvic pain and subfertility or infertility, which profoundly affect the quality of life in women. Current hormonal therapies to induce a hypoestrogenic state are unsuccessful because of undesirable side effects, reproductive health concerns, and failure to prevent recurrence of disease. There is a fundamental need to identify nonestrogen or nonsteroidal targets for the treatment of endometriosis. Peritoneal fluid concentrations of prostaglandin E2 (PGE2) are higher in women with endometriosis, and this increased PGE2 plays important role in survival and growth of endometriosis lesions. The objective of the present study was to determine the effects of pharmacological inhibition of PGE2 receptors, EP2 and EP4, on molecular and cellular aspects of the pathogenesis of endometriosis and associated clinical symptoms. Using human fluorescent endometriotic cell lines and chimeric mouse model as preclinical testing platform, our results, to our knowledge for the first time, indicate that selective inhibition of EP2/EP4: (i) decreases growth and survival of endometriosis lesions; (ii) decreases angiogenesis and innervation of endometriosis lesions; (iii) suppresses proinflammatory state of dorsal root ganglia neurons to decrease pelvic pain; (iv) decreases proinflammatory, estrogen-dominant, and progesterone-resistant molecular environment of the endometrium and endometriosis lesions; and (v) restores endometrial functional receptivity through multiple mechanisms. Our novel findings provide a molecular and preclinical basis to formulate long-term nonestrogen or nonsteroidal therapy for endometriosis.
Assuntos
Endometriose/tratamento farmacológico , Endometriose/patologia , Receptores de Prostaglandina E Subtipo EP2/antagonistas & inibidores , Receptores de Prostaglandina E Subtipo EP4/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Compostos de Bifenilo/farmacologia , Compostos de Bifenilo/uso terapêutico , Caspase 3/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Endométrio/irrigação sanguínea , Endométrio/patologia , Estrogênios/biossíntese , Feminino , Humanos , Inflamação/patologia , Camundongos , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Dor Pélvica/tratamento farmacológico , Dor Pélvica/patologia , Poli(ADP-Ribose) Polimerases/metabolismo , Progesterona/metabolismo , Transdução de Sinais/efeitos dos fármacos , Esteroides/uso terapêutico , Xantonas/farmacologia , Xantonas/uso terapêuticoRESUMO
Resveratrol (RVT), a polyphenolic component in grapes and red wine, has been known for its cytoprotective actions against several diseases. However, beneficial effects of RVT against early exposure to endocrine disrupting chemicals (EDCs) have not been understood. EDCs are linked to several ovarian diseases such as premature ovarian failure, polycystic ovary syndrome, early menopause and infertility in women. Hexavalent chromium (CrVI) is a heavy metal EDC, and widely used in >50 industries. Environmental contamination with CrVI in the US is rapidly increasing, predisposing the human to several illnesses including cancers and still birth. Our lab has been involved in determining the molecular mechanism of CrVI-induced female infertility and intervention strategies to mitigate CrVI effects. Lactating mother rats were exposed to CrVI (50ppm potassium dichromate) from postpartum days 1-21 through drinking water with or without RVT (10mg/kg body wt., through oral gavage daily). During this time, F1 females received respective treatments through mother's milk. On postnatal day (PND) 25, blood and the ovary, kidney and liver were collected from the F1 females for analyses. CrVI increased atresia of follicles by increasing cytochrome C and cleaved caspase-3; decreasing antiapoptotic proteins; decreasing estradiol (E2) biosynthesis and enhancing metabolic clearance of E2, increasing oxidative stress and decreasing endogenous antioxidants. RVT mitigated the effects of CrVI by upregulating cell survival proteins and AOXs; and restored E2 levels by inhibiting hydroxylation, glucuronidation and sulphation of E2. This is the first study to report the protective effects of RVT against any toxicant in the ovary.
Assuntos
Cromo/toxicidade , Ovário/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Estilbenos/farmacologia , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Carcinógenos/toxicidade , Catalase/metabolismo , Poluentes Ambientais/toxicidade , Estradiol/sangue , Estradiol/metabolismo , Feminino , Atresia Folicular/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Oócitos/efeitos dos fármacos , Oócitos/patologia , Ovário/patologia , Gravidez , Progesterona/sangue , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Resveratrol , Superóxido Dismutase/metabolismo , Testosterona/sangueRESUMO
Hexavalent chromium (CrVI), one of the more toxic heavy metals, is widely used in more than 50 industries such as chrome plating, welding, wood processing and tanneries. As one of the world's leading producers of chromium compounds, the U.S. is facing growing challenges in protecting human health against multiple adverse effects of CrVI. CrVI is rapidly converted to CrIII intracellularly, and can induce apoptosis through different mechanisms. Our previous studies demonstrated postnatal exposure to CrVI results in a delay or arrest in follicle development and puberty. Pregnant rats were treated with 25 ppm potassium dichromate (CrVI) from gestational day (GD) 9.5 to 14.5 through drinking water, placentae were removed on GD 20, and total Cr was estimated in the placentae; ovaries were removed from the F1 offspring on postnatal day (PND)-1 and various analyses were performed. Our results show that gestational exposure to CrVI resulted in (i) increased Cr concentration in the placenta, (ii) increased germ cell apoptosis by up-regulating p53/p27-Bax-caspase-3 proteins and by increasing p53-SOD-2 co-localization; (iii) accelerated germ cell cyst (GCC) breakdown; (iv) advanced primordial follicle assembly and primary follicle transition and (v) down regulation of p-AKT, p-ERK and XIAP. As a result of the above events, CrVI induced early reproductive senescence and decrease in litter size in F1 female progeny.
Assuntos
Apoptose , Cromo/toxicidade , Células Germinativas/efeitos dos fármacos , Células Germinativas/patologia , Exposição Materna/efeitos adversos , Reprodução/efeitos dos fármacos , Animais , Cistos/metabolismo , Feminino , Regulação da Expressão Gênica , Imuno-Histoquímica , Microscopia de Fluorescência , Folículo Ovariano/efeitos dos fármacos , Ovário/efeitos dos fármacos , Placenta/efeitos dos fármacos , Dicromato de Potássio/química , Gravidez , Prenhez , Ratos , Proteína Supressora de Tumor p53/metabolismoRESUMO
Environmental exposure to endocrine-disrupting chemicals (EDCs) is one cause of premature ovarian failure (POF). Hexavalent chromium (CrVI) is a heavy metal EDC widely used in more than 50 industries, including chrome plating, welding, wood processing, and tanneries. Recent data from U.S. Environmental Protection Agency indicate increased levels of Cr in drinking water from several American cities, which potentially predispose residents to various health problems. Recently, we demonstrated that gestational exposure to CrVI caused POF in F1 offspring. The current study was performed to identify the molecular mechanism behind CrVI-induced POF. Pregnant rats were treated with 25 ppm of potassium dichromate from Gestational Day (GD) 9.5 to GD 14.5 through drinking water, and the fetuses were exposed to CrVI through transplacental transfer. Ovaries were removed from the fetuses or pups on Embryonic Day (ED) 15.5, ED 17.5, Postnatal Day (PND) 1, PND 4, or PND 25, and various analyses were performed. Results showed that gestational exposure to CrVI: 1) increased germ cell/oocyte apoptosis and advanced germ cell nest (GCN) breakdown; 2) increased X-prolyl aminopeptidase (Xpnpep) 2, a POF marker in humans, during GCN breakdown; 3) decreased Xpnpep2 during postnatal follicle development; and 4) increased colocalization of Xpnpep2 with Col3 and Col4. We also found that Xpnpep2 inversely regulated the expression of Col1, Col3, and Col4 in all the developmental stages studied. Thus, CrVI advanced GCN breakdown and increased follicle atresia in F1 female progeny by targeting Xpnpep2.
Assuntos
Aminopeptidases/fisiologia , Cromo/efeitos adversos , Cromo/farmacologia , Fase Folicular/efeitos dos fármacos , Óvulo/efeitos dos fármacos , Insuficiência Ovariana Primária/induzido quimicamente , Insuficiência Ovariana Primária/fisiopatologia , Animais , Apoptose/efeitos dos fármacos , Carcinógenos Ambientais/efeitos adversos , Carcinógenos Ambientais/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Colágeno Tipo I/fisiologia , Colágeno Tipo III/fisiologia , Colágeno Tipo IV/fisiologia , Modelos Animais de Doenças , Feminino , Atresia Folicular/efeitos dos fármacos , Atresia Folicular/fisiologia , Fase Folicular/fisiologia , Ovário/efeitos dos fármacos , Ovário/fisiologia , Óvulo/fisiologia , Gravidez , RatosRESUMO
Prenatal exposure to endocrine disrupting chemicals (EDCs), including bisphenol A, dioxin, pesticides, and cigarette smoke, has been linked to several ovarian diseases such as premature ovarian failure (POF) and early menopause in women. Hexavalent chromium (CrVI), one of the more toxic heavy metals, is widely used in more than 50 industries. As one of the world's leading producers of Cr compounds, the U.S. is facing growing challenges in protecting human health against adverse effects of CrVI. Our recent findings demonstrated that in vivo CrVI exposure during gestational period caused POF in F1 offspring. Our current research focus is three-fold: (i) to identify the effect of CrVI on critical windows of great vulnerability of fetal ovarian development; (ii) to understand the molecular mechanism of CrVI-induced POF; (iii) to identify potential intervention strategies to mitigate or inhibit CrVI effects. In order to accomplish these goals we used a fetal whole ovarian culture system. Fetuses were removed from the normal pregnant rats on gestational day 13.5. Fetal ovaries were cultured in vitro for 12 days, and treated with or without 0.1 ppm potassium dichromate (CrVI) from culture day 2-8, which recapitulated embryonic day 14.5-20.5, in vivo. Results showed that CrVI increased germ cell/oocyte apoptosis by increasing caspase 3, BAX, p53 and PUMA; decreasing BCL2, BMP15, GDF9 and cKIT; and altering cell cycle regulatory genes and proteins. This model system may serve as a potential tool for high throughput testing of various drugs and/or EDCs in particular to assess developmental toxicity of the ovary.
Assuntos
Cromo/toxicidade , Feto/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Ovário/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Proteína Morfogenética Óssea 15/genética , Proteína Morfogenética Óssea 15/metabolismo , Caspase 3/genética , Caspase 3/metabolismo , Relação Dose-Resposta a Droga , Feminino , Fator 9 de Diferenciação de Crescimento/genética , Fator 9 de Diferenciação de Crescimento/metabolismo , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Técnicas de Cultura de Órgãos , Dicromato de Potássio/toxicidade , Gravidez , Ratos , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
In ruminants, prostaglandin F2 alpha (PGF2alpha) is synthesized and released in a pulsatile pattern from the endometrial luminal epithelial (LE) cells during the process of luteolysis. Interferon tau (IFNT) is a Type 1 IFN secreted by the trophoblast cells of the developing conceptus. IFNT acts locally on endometrial LE cells to inhibit pulsatile releases of PGF2alpha and thus establish an endocrine environment for recognition of pregnancy. Cell signaling pathways through which IFNT stimulates expression of multiple genes or proteins in endometrial LE are largely unknown. Results of the present investigation indicate that intrauterine administration of IFNT inhibits pulsatile release of PGF2alpha, while coadministration IFNT and ERK 1/2 inhibitor U0126 restores luteolytic PGF2alpha pulses in sheep. IFNT increases phosphorylation of ERK1/2 proteins and increases its interaction with PGT proteins in endometrial LE. Blockade of ERK1/2 pathways inhibits IFNT action, decreases pERK1/2 and PGT protein interactions, and re-establishes the spatial expression of the oxytocin receptor protein completely and the estrogen receptor protein partially without modulating the expression of interferon regulatory factor-2 (IRF-2) protein in endometrial LE. IFNT does not decrease expression of COX-2, PGDH, or PGT protein in endometrial LE. Our results provide important new insights into IFNT signaling and the molecular endocrine control of PGF2alpha release at the time of establishment of pregnancy in ruminants. This novel IFNT-ERK1/2 signaling module needs to be explored in future studies to understand molecular and cellular mechanisms of IFNT action in endometrial LE in ruminants.
Assuntos
Butadienos/farmacologia , Dinoprosta/metabolismo , Endométrio/efeitos dos fármacos , Interferon Tipo I/farmacologia , Luteólise/fisiologia , Nitrilas/farmacologia , Proteínas da Gravidez/farmacologia , Animais , Endométrio/metabolismo , Epitélio/efeitos dos fármacos , Epitélio/fisiologia , Ciclo Estral/efeitos dos fármacos , Ciclo Estral/fisiologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Interferon Tipo I/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Gravidez , Proteínas da Gravidez/metabolismoRESUMO
Environmental contamination of drinking water with chromium (Cr) has been increasing in more than 30 cities in the United States. Previous studies from our group have shown that Cr affects reproductive functions in female Sprague Dawley rats. Although it is impossible to completely remove Cr from the drinking water, it is imperative to develop effective intervention strategies to inhibit Cr-induced deleterious health effects. Edaravone (EDA), a potential inhibitor of free radicals, has been clinically used to treat cancer and cardiac ischemia. This study evaluated the efficacy of EDA against Cr-induced ovarian toxicity. Results showed that maternal exposure to CrVI in rats increased follicular atresia, decreased steroidogenesis, and delayed puberty in F1 offspring. CrVI increased oxidative stress and decreased antioxidant (AOX) enzyme levels in the ovary. CrVI increased follicle atresia by increased expression of cleaved caspase 3, and decreased expression of Bcl2 and Bcl2l1 in the ovary. EDA mitigated or inhibited the effects of CrVI on follicle atresia, pubertal onset, steroid hormone levels, and AOX enzyme activity, as well as the expression of Bcl2 and Bcl2l1 in the ovary. In a second study, CrVI treatment was withdrawn, and F1 rats were injected with estradiol (E2) (10 µg in PBS/ethanol per 100 g body weight) for a period of 2 wk to evaluate whether E2 treatment will restore Cr-induced depletion of AOX enzymes. E2 restored CrVI-induced depletion of glutathione peroxidase 1, catalase, thioredoxin 2, and peroxiredoxin 3 in the ovary. This is the first study to demonstrate the protective effects of EDA against any toxicant in the ovary.
Assuntos
Estrogênios/uso terapêutico , Sequestradores de Radicais Livres/uso terapêutico , Intoxicação por Metais Pesados , Ovário/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Oxirredutases/biossíntese , Intoxicação/prevenção & controle , Poluentes Químicos da Água/antagonistas & inibidores , Animais , Animais Lactentes , Antipirina/administração & dosagem , Antipirina/análogos & derivados , Antipirina/uso terapêutico , Apoptose/efeitos dos fármacos , Relação Dose-Resposta a Droga , Edaravone , Estradiol/administração & dosagem , Estradiol/uso terapêutico , Estrogênios/administração & dosagem , Feminino , Sequestradores de Radicais Livres/administração & dosagem , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Infertilidade Feminina/etiologia , Infertilidade Feminina/prevenção & controle , Injeções Intraperitoneais , Lactação , Troca Materno-Fetal , Ovário/enzimologia , Ovário/patologia , Oxirredutases/antagonistas & inibidores , Intoxicação/tratamento farmacológico , Intoxicação/enzimologia , Intoxicação/fisiopatologia , Dicromato de Potássio/administração & dosagem , Dicromato de Potássio/antagonistas & inibidores , Dicromato de Potássio/toxicidade , Gravidez , Ratos Sprague-Dawley , Poluentes Químicos da Água/administração & dosagem , Poluentes Químicos da Água/toxicidadeRESUMO
Hexavalent chromium (CrVI) is a highly toxic metal and a major environmental pollutant. Several studies indicate that CrVI exposure adversely affects reproductive function. We reported that maternal Cr exposure resulted in Cr accumulation in the reproductive organs of female offsprings. CrVI can cross the placental barrier and also can be passed through breastfeeding. The present investigation aimed to determine the persistent (in utero through puberal period) CrVI exposure-induced toxic effects on the reproductive functions of mother and the offspring. Induction of oxidative stress is one of the plausible mechanisms behind Cr-induced cellular deteriorations. Mother rats exposed to CrVI showed reduced reproductive outcome, while the offsprings showed higher accumulation of Cr in ovary, altered steroid, and peptide hormones. Specific activities of antioxidant enzymes were decreased and associated with increased levels of H2 O2 , and lipid peroxidation. CrVI exposure also damaged the ovarian histoarchitecture in various age groups studied. CrVI exposure also delayed the sexual maturation. Results from the present investigation suggest that CrVI exposure from in utero through puberal period significantly damaged the pubertal development through altered antioxidants, anemia, and altered hormone levels. These changes were associated with damaged ovarian histoarchitecture and extended estrous cycle in developing Wistar rats.
Assuntos
Cromo/toxicidade , Poluentes Ambientais/toxicidade , Animais , Antioxidantes/metabolismo , Feminino , Hormônios Gonadais/sangue , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Exposição Materna/efeitos adversos , Troca Materno-Fetal , Ovário/citologia , Ovário/efeitos dos fármacos , Ovário/crescimento & desenvolvimento , Estresse Oxidativo/efeitos dos fármacos , Hormônios Peptídicos/sangue , Gravidez , Ratos , Ratos Wistar , Reprodução/efeitos dos fármacos , Maturidade Sexual/efeitos dos fármacosRESUMO
Hexavalent chromium, Cr(VI), is a heavy metal endocrine disruptor used widely in various industries worldwide and is considered a reproductive toxicant. Our previous studies demonstrated that lactational exposure to Cr(VI) caused follicular atresia, disrupted steroid hormone biosynthesis and signaling, and delayed puberty. However, the underlying mechanism was unknown. The current study investigated the effects of Cr(VI) exposure (25 ppm) during postnatal days 1-21 via dam's milk on epigenetic alterations in the ovary of F1 offspring. Data indicated that Cr(VI) disrupted follicle development and caused apoptosis by increasing DNMT3a /3b and histone methyl marks (H3K27me3 and H3K9me3) along with decreasing histone acetylation marks (H3K9ac and H3K27ac). Our study demonstrates that exposure to Cr(VI) causes changes in the epigenetic marks, partially contributing to the transcriptional repression of genes regulating ovarian development, cell proliferation (PCNA), cell survival (BCL-XL and BCL-2), and activation of genes regulating apoptosis (AIF and cleaved caspase-3), resulting in follicular atresia. The current study suggests a role for epigenetics in Cr(VI)-induced ovotoxicity and infertility.
Assuntos
Histonas , Ovário , Feminino , Humanos , Atresia Folicular , Cromo/toxicidade , Apoptose , Epigênese GenéticaRESUMO
In ruminants, endometrial prostalgandin (PG) F(2alpha) causes functional luteolysis, whereas luteal synthesis of PGF(2alpha) is required for structural luteolysis. PGE(2) is considered to be a luteoprotective mediator. Molecular aspects of luteal PGF(2alpha) and PGE(2) biosynthesis and signaling during the estrous cycle and establishment of pregnancy are largely unknown. The objectives of the present study were 1) to determine the regulation of proteins involved in PGF(2alpha) and PGE(2) biosynthesis, catabolism, transport and signaling in the corpus luteum (CL); 2) to investigate the transport of interferon tau (IFNT), PGF(2alpha), and PGE(2) from the uterus to the ovary through the vascular utero-ovarian plexus (UOP); and 3) to compare the intraluteal production of PGF(2alpha) and PGE(2) on Days 12, 14, and 16 of the estrous cycle and pregnancy in sheep. Our results indicate that luteal PG biosynthesis is selectively directed towards PGF(2alpha) at the time of luteolysis and towards PGE(2) during the establishment of pregnancy. Moreover, the ability of the CL of early pregnancy to resist luteolysis is due to increased intraluteal biosynthesis of PGE(2) and PGE(2) receptor (PTGER) 2 (also known as EP2)- and PTGER4 (also known as EP4)-mediated signaling. We also found that IFNT protein is not transported through the UOP from the uterus to the ovary; in contrast, a large proportion of endometrial PGE(2) is transported from the uterus to the ovary through the UOP. These results indicate that endometrial PGE(2) stimulated by pregnancy is transported locally to the ovary, which increases luteal PGE(2) biosynthesis and hence activates luteal PTGER2 and PTGER4 signaling, thus protecting the CL during the establishment of pregnancy in sheep.
Assuntos
Dinoprosta/metabolismo , Dinoprostona/metabolismo , Células Lúteas/metabolismo , Luteólise/metabolismo , Prenhez , Prostaglandinas/biossíntese , Ovinos , Animais , Ciclo Estral/metabolismo , Ciclo Estral/fisiologia , Feminino , Histerectomia/veterinária , Luteólise/fisiologia , Masculino , Gravidez , Prenhez/metabolismo , Prostaglandinas/metabolismo , Distribuição Aleatória , Receptores de Prostaglandina/metabolismo , Ovinos/metabolismo , Ovinos/fisiologia , Transdução de Sinais/fisiologia , Especificidade por SubstratoRESUMO
Hexavalent chromium (CrVI) is a highly toxic metal and major environmental pollutant and is extensively used in more than 50 industries. The major route of CrVI exposure for the general population is oral intake. Chromium is considered an important nutrient responsible for carbohydrate metabolism. However, excess CrVI exposure is associated with various pathological conditions including reproductive dysfunction. CrVI can traverse the placental barrier and cause wide range of abnormalities in fetal development. Cr is transported to offspring through mother's milk in lactating women exposed to CrVI. Therefore, the present study was carried out to determine the toxic effects of lactational CrVI exposure on ovary and uterus and the beneficial role of vitamin C in preventing/ameliorating the toxic effects of CrVI in developing female Wistar rats. Generation of oxidative stress is considered one of the plausible mechanisms behind Cr-induced cellular deteriorations. The present study evidenced a decrease in the specific activities of antioxidants, serum testosterone and progesterone and an increase in the levels of H2O2, lipid peroxidation (LPO) and follicle stimulating hormone in rats exposed to CrVI when compared to control. CrVI exposure also delayed the sexual maturation and extended the estrous cycle. Simultaneous administration of vitamin C significantly prevented the increase in LPO and enhanced the antioxidant status. These results suggest the protective effect of vitamin C against the CrVI exposure-induced toxicity and attest the significance of antioxidants in diet.
Assuntos
Ácido Ascórbico/farmacologia , Cromo/toxicidade , Ovário/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Maturidade Sexual/efeitos dos fármacos , Útero/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Peso Corporal , Cromo/análise , Cromo/sangue , Ciclo Estral , Feminino , Hormônio Foliculoestimulante/sangue , Peróxido de Hidrogênio , Lactação , Peroxidação de Lipídeos/efeitos dos fármacos , Hormônio Luteinizante/sangue , Exposição Materna , Tamanho do Órgão , Ovário/química , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Oxirredutases/metabolismo , Gravidez , Ratos , Ratos Wistar , Útero/química , Útero/crescimento & desenvolvimento , Útero/metabolismoRESUMO
Environmental contamination with hexavalent chromium, Cr(VI), has been increasing in the United States as well as in developing countries. Exposure to Cr(VI) predisposes the human population to various diseases, including cancer, infertility, and developmental problems in children. Previous findings from our laboratory reported that prenatal exposure to Cr(VI) caused premature ovarian failure through p53-mediated mechanisms. Sirtuin 1 (SIRT1) is an NAD+ -dependent histone deacetylase class III. SIRT1 deacetylates several histones and non-histone proteins such as p53 and NFkB. The current study determines a role for the SIRT1-p53 network in apoptosis induced by Cr(VI) in the ovary and establishes physical interaction between SIRT1 and p53. Adult pregnant dams were given regular drinking water or Cr(VI) (10 ppm potassium dichromate in drinking water, ad libitum), and treated with SIRT1 inhibitor, EX-527 (50 mg/kg body weight, i.p.,), during 9.5 - 14.5 days post-coitum. On postnatal day-1, ovaries from F1 offspring were collected for various analyses. Results indicated that Cr(VI) increased germ cell and somatic cell apoptosis, upregulated acetyl-p53, activated the apoptotic pathway, and inhibited cell survival pathways. Cr(VI) decreased acetyl-p53-SIRT1 co-localization in the ovary. In an immortalized rat granulosa cell line SIGC, Cr(VI) inhibited the physical interaction between SIRT1 and acetyl-p53 by altering the p53:SIRT1 ratio. EX-527 exacerbated Cr(VI)-induced mechanisms. The current study shows a novel mechanism for Cr(VI)-induced apoptosis in the ovary, mediated through the p53-SIRT1 network, suggesting that targeting the p53 pathway may be an ideal approach to rescue ovaries from Cr(VI)-induced apoptosis.
Assuntos
Ovário , Sirtuína 1 , Animais , Apoptose , Cromo/toxicidade , Feminino , Ovário/metabolismo , Gravidez , Ratos , Sirtuína 1/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
Hexavalent chromium (CrVI) has been widely used in industries throughout the world. Increased usage of CrVI and atmospheric emission of CrVI from catalytic converters of automobiles, and its improper disposal causes various health hazards including female infertility. Recently we have reported that lactational exposure to CrVI induced a delay/arrest in follicular development at the secondary follicular stage. In order to investigate the underlying mechanism, primary cultures of rat granulosa cells were treated with 10 µM potassium dichromate (CrVI) for 12 and 24h, with or without vitamin C pre-treatment for 24h. The effects of CrVI on intrinsic apoptotic pathway(s) were investigated. Our data indicated that CrVI: (i) induced DNA fragmentation and increased apoptosis, (ii) increased cytochrome c release from the mitochondria to cytosol, (iii) downregulated anti-apoptotic Bcl-2, Bcl-XL, HSP70 and HSP90; upregulated pro-apoptotic BAX and BAD, (iv) altered translocation of Bcl-2, Bcl-XL, BAX, BAD, HSP70 and HSP90 to the mitochondria, (v) upregulated p-ERK and p-JNK, and selectively translocated p-ERK to the mitochondria and nucleus, (vi) activated caspase-3 and PARP, and (vii) increased phosphorylation of p53 at ser-6, ser-9, ser-15, ser-20, ser-37, ser-46 and ser-392, increased p53 transcriptional activation, and downregulated MDM-2. Vitamin C pre-treatment mitigated CrVI effects on apoptosis and related pathways. Our study, for the first time provides a clear insight into the effect of CrVI on multiple pathways that lead to apoptosis of granulosa cells which could be mitigated by vitamin C.
Assuntos
Apoptose/efeitos dos fármacos , Ácido Ascórbico/farmacologia , Cromo/toxicidade , Fragmentação do DNA/efeitos dos fármacos , Células da Granulosa/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Regulação para Baixo/efeitos dos fármacos , Feminino , Células da Granulosa/metabolismo , Proteína Quinase 1 Ativada por Mitógeno , Proteína Quinase 3 Ativada por Mitógeno , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima/efeitos dos fármacosRESUMO
Particulate matter (PM) causes adverse developmental outcomes following prenatal exposure, but the underlying biological mechanisms remain uncertain. Here we elucidate the effects of diesel exhaust ultrafine particle (UFP) exposure during pregnancy on placental and fetal development. Time-mated C57Bl/6n mice were gestationally exposed to UFPs at a low dose (LD, 100 µg/m3) or high dose (HD, 500 µg/m3) for 6 h daily. Phenotypic effects on fetuses and placental morphology at gestational day (GD) of 18.5 were evaluated, and RNA sequencing was characterized for transcriptomic changes in placental tissue from male and female offspring. A significant decrease in average placental weights and crown to rump lengths was observed in female offspring in the LD exposure group. Gestational UFP exposure altered placental morphology in a dose- and sex-specific manner. Average female decidua areas were significantly greater in the LD and HD groups. Maternal lacunae mean areas were increased in the female LD group, whereas fetal blood vessel mean areas were significantly greater in the male LD and HD groups. RNA sequencing indicated several disturbed cellular functions related to lipid metabolism, which were most pronounced in the LD group and especially in female placental tissue. Our findings demonstrate the vulnerability of offspring exposed to UFPs during pregnancy, highlighting sex-specific effects and emphasizing the importance of mitigating PM exposure to prevent adverse health outcomes.
Assuntos
Material Particulado , Efeitos Tardios da Exposição Pré-Natal , Animais , Feminino , Redes Reguladoras de Genes , Masculino , Camundongos , Material Particulado/toxicidade , Placenta , Gravidez , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Emissões de Veículos/toxicidadeRESUMO
Hormones are known to influence various body systems that include skeletal, cardiac, digestive, excretory, and immune systems. Emerging investigations suggest the key role played by secretions of endocrine glands in immune cell differentiation, proliferation, activation, and memory attributes of the immune system. The link between steroid hormones such as glucocorticoids and inflammation is widely known. However, the role of peptide hormones and amino acid derivatives such as growth and thyroid hormones, prolactin, dopamine, and thymopoietin in regulating the functioning of the immune system remains unclear. Here, we reviewed the findings pertinent to the functional role of hormone-immune interactions in health and disease and proposed perspective directions for translational research in the field.
Assuntos
Doenças do Sistema Endócrino/metabolismo , Sistema Endócrino/metabolismo , Hormônio do Crescimento/metabolismo , Doenças do Sistema Imunitário/metabolismo , Sistema Imunitário/metabolismo , Prolactina/metabolismo , Timócitos/metabolismo , Animais , Comunicação Celular , Citocinas/genética , Citocinas/imunologia , Citocinas/metabolismo , Dopamina/genética , Dopamina/imunologia , Dopamina/metabolismo , Sistema Endócrino/citologia , Sistema Endócrino/imunologia , Doenças do Sistema Endócrino/genética , Doenças do Sistema Endócrino/imunologia , Doenças do Sistema Endócrino/patologia , Glucocorticoides/genética , Glucocorticoides/imunologia , Glucocorticoides/metabolismo , Hormônio do Crescimento/genética , Hormônio do Crescimento/imunologia , Humanos , Sistema Imunitário/citologia , Sistema Imunitário/imunologia , Doenças do Sistema Imunitário/genética , Doenças do Sistema Imunitário/imunologia , Doenças do Sistema Imunitário/patologia , Lactotrofos/citologia , Lactotrofos/imunologia , Lactotrofos/metabolismo , Prolactina/genética , Prolactina/imunologia , Receptores Dopaminérgicos/genética , Receptores Dopaminérgicos/imunologia , Receptores Dopaminérgicos/metabolismo , Somatotrofos/citologia , Somatotrofos/imunologia , Somatotrofos/metabolismo , Timócitos/citologia , Timócitos/imunologia , Hormônios Tireóideos/genética , Hormônios Tireóideos/imunologia , Hormônios Tireóideos/metabolismoRESUMO
Major limitations in understanding the direct effects of endocrine-disrupting chemicals (EDCs) and cell signaling events in ovarian cellular dynamics in mammals include a lack of proper and simple tools/techniques as well as gaps in knowledge regarding the critical window(s) of vulnerability. Identifying and validating such tools and evaluating the effects of EDCs on molecular dynamics and cellular events during the critical windows of ovarian development are very important to improve the fertility in women and preserve the future health of the developing fetuses. Therefore, we developed a fetal whole ovarian ex vivo culture model. Ex vivo ovary culture models allow varying culture parameters in a highly controlled manner and thus have the potential to allow a more thorough evaluation for reproductive toxicity studies and drug response. This chapter describes clear and thorough details for setting up and maintaining an ex vivo culture system from the rat ovaries and further analyses of mRNA and protein expressions and estimating follicle numbers.
Assuntos
Técnicas de Cultura de Células/métodos , Disruptores Endócrinos/toxicidade , Ovário/citologia , Ovário/embriologia , Animais , Células Cultivadas , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Modelos Biológicos , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Ovário/efeitos dos fármacos , Ovário/metabolismo , Ratos , ReproduçãoRESUMO
Environmental contamination with hexavalent chromium (CrVI) is a growing problem both in the United States and developing countries. Hexavalent chromium is widely used in numerous industries. Environmental exposure to CrVI adversely affects pregnancy outcomes and subsequent health of 2 generations, resulting in higher pregnancy loss, spontaneous abortion and low birth rate. Pregnant women exposed to CrVI through occupational settings experience increased risk of spontaneous abortion, stillbirth, preterm birth, and neonatal death. Children of the CrVI exposed women experience respiratory problems, perinatal jaundice, and increased birth defects. Because placental dysfunction may have a role in such adverse pregnancy outcome, we tested the hypothesis that environmental Cr exposure in pregnant women results in Cr accumulation in the human placenta, which could increase placental oxidative stress by disrupting antioxidant machinery and inducing apoptosis. Studies using frozen, deidentified human term placenta samples indicated that: (1) Cr accumulates in human term placenta tissues and (2) increase in Cr accumulation is positively correlated with oxidative stress and apoptotic markers, and altered antioxidants levels. Interestingly, there was a sexual dimorphism in the correlation between Cr accumulation and oxidative stress, and expression of apoptotic and antioxidant markers. Mechanistic in vitro studies using human trophoblast cells BeWo confirmed the detrimental effects of Cr in altering antioxidant genes. For the first time, this study provides evidence in support of a positive correlation between Cr accumulation in the human placenta and accelerated oxidative stress, with a gender bias toward the male sex.
Assuntos
Apoptose/efeitos dos fármacos , Cromo/toxicidade , Poluentes Ambientais/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Placenta/efeitos dos fármacos , Caracteres Sexuais , Adulto , Antioxidantes/metabolismo , Apoptose/genética , Biomarcadores/metabolismo , Linhagem Celular , Cromo/metabolismo , Feminino , Humanos , Técnicas In Vitro , Recém-Nascido , Masculino , Estresse Oxidativo/genética , Placenta/metabolismo , Placenta/patologia , Gravidez , Trofoblastos/efeitos dos fármacos , Trofoblastos/metabolismo , Trofoblastos/patologia , Adulto JovemRESUMO
Environmental contamination with hexavalent chromium (CrVI) is a growing problem both in the U.S and developing countries. CrVI is a heavy-metal endocrine disruptor; women working in Cr industries exhibit an increased incidence of premature abortion and infertility. The current study was designed to understand the mechanism of CrVI toxicity on placental cell survival/death pathways. Pregnant mothers were treated with or without CrVI (50ppmK2Cr2O7) through drinking water from gestational day (GD) 9.5-14.5, and placentas were analyzed on GD 18.5. Results indicated that CrVI increased apoptosis of trophoblasts, vascular endothelium of the metrial glands and yolk sac epithelium through caspase-3 and p53-dependent pathways. CrVI increased apoptosis in labyrinth and basal zones in a caspase-3-independent manner via AIF, and through an ATM-p53-NOXA-PUMA-p27 network. CrVI downregulated cell survival proteins Bcl-2, Bcl-XL and XIAP in the placenta. CrVI disrupts placental histoarchitecture and increases cell death by spatiotemporal modulation of apoptotic signaling.
Assuntos
Apoptose/efeitos dos fármacos , Cromo/toxicidade , Disruptores Endócrinos/toxicidade , Poluentes Ambientais/toxicidade , Placenta/efeitos dos fármacos , Animais , Sobrevivência Celular/efeitos dos fármacos , Feminino , Idade Gestacional , Exposição Materna , Placenta/metabolismo , Placenta/patologia , Gravidez , Ratos Sprague-Dawley , Análise Espaço-TemporalRESUMO
Epidemiologic studies document relationships between chromium VI (CrVI) exposure and increased risk of spontaneous abortion, stillbirth, preterm birth, and neonatal death in pregnant women. Environmental contamination with CrVI is a growing problem both in the United States and developing countries. CrVI is widely used in numerous industries. This study was designed to understand the mechanism of CrVI toxicity on placental oxidative stress and antioxidant (AOX) machinery. Pregnant mother rats were treated with or without CrVI (50 ppm K2Cr2O7) through drinking water from gestational day (GD) 9.5-14.5, and placentas were analyzed on GD 18.5. Results indicated that CrVI reduced the trophoblast cell population. CrVI increased reactive oxygen species (ROS) and decreased the expression of AOX proteins. CrVI disrupts the trophoblast proliferation of the placenta. This study provides insight into the critical role of AOXs in placental function.