RESUMO
The intrauterine environment is particularly vulnerable to environmental exposures. We previously established a mouse model that provided evidence for pregnancy complications and placental anti-angiogenesis in response to Aroclor 1254 (A-1254), a mixture of polychlorinated biphenyls (PCBs). Importantly, these effects were observed in IL-10-/-, but not wild type, mice, suggesting that IL-10 deficiency predisposes to pregnancy disruptive effects of environmental toxicants. However, the mechanisms by which PCBs cause anti-angiogenic effects are unclear. Here, we evaluated PCB-mediated anti-angiogenic effects by diverse but complementary approaches, including HUVEC-mediated trophoblast invasion in nude mice, in vitro three-dimensional capillary tube formation involving HUVEC and/or HTR8 trophoblasts, and aortic ring endothelial cell outgrowth/sprouting. Taken together, our data suggest that PCBs act as potent anti-angiogenic agents. Importantly, we show that treatment of pregnant IL-10-/- mice with A-1254 resulted in placental activation of the Notch/Delta-like ligand (Dll) pathway, a master regulator of cell-cell interaction and vascular patterning. Similar results were obtained with HUVEC and HTR8 trophoblasts. Rescue of A-1254-induced disruption of HUVEC-based tube formation by γ-secretase inhibitor L1790 confirmed the critical role of the Notch/Dll pathway. Our data suggest that PCBs impart pregnancy disruptive functions by activating the Notch/Dll pathway and by inducing anti-angiogenic effects at the maternal-fetal interface.
Assuntos
Inibidores da Angiogênese/toxicidade , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas de Membrana/metabolismo , Bifenilos Policlorados/toxicidade , Complicações na Gravidez/metabolismo , Receptor Notch4/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Animais , Proteínas de Ligação ao Cálcio , Modelos Animais de Doenças , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Interleucina-10/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Gravidez , Complicações na Gravidez/induzido quimicamente , Transdução de SinaisRESUMO
Preeclampsia is a major pregnancy complication with potential short- and long-term consequences for both mother and fetus. Understanding its pathogenesis and causative biomarkers is likely to yield insights for prediction and treatment. Herein, we provide evidence that transthyretin, a transporter of thyroxine and retinol, is aggregated in preeclampsia and is present at reduced levels in sera of preeclamptic women, as detected by proteomic screen. We demonstrate that transthyretin aggregates form deposits in preeclampsia placental tissue and cause apoptosis. By using in vitro approaches and a humanized mouse model, we provide evidence for a causal link between dysregulated transthyretin and preeclampsia. Native transthyretin inhibits all preeclampsia-like features in the humanized mouse model, including new-onset proteinuria, increased blood pressure, glomerular endotheliosis, and production of anti-angiogenic factors. Our findings suggest that a focus on transthyretin structure and function is a novel strategy to understand and combat preeclampsia.
Assuntos
Pré-Eclâmpsia/metabolismo , Pré-Eclâmpsia/patologia , Pré-Albumina/metabolismo , Animais , Modelos Animais de Doenças , Endoglina , Feminino , Humanos , Imunoprecipitação , Interleucina-10/deficiência , Interleucina-10/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Camundongos , Neovascularização Fisiológica , Pré-Eclâmpsia/sangue , Pré-Albumina/química , Gravidez , Ligação Proteica , Estrutura Quaternária de Proteína , Proteômica , Solubilidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Recurrent spontaneous abortion (RSA), three or more pregnancy losses prior to 20 weeks, occurs in about 1% of all pregnancies, 50% of RSA cases remain unexplained and unresolved. Recently, immune pathways have been implicated in the pathophysiology of RSA. Immune tolerance of the fetal-placental unit and placental angiogenesis are mandatory for a successful pregnancy outcome. Unscheduled dysregulation of the placental vasculature is thought to be the pathophysiologic mechanisms underlying an array of pregnancy complications like infertility, miscarriage, pre-eclampsia, and fetal growth restriction and death. Investigations on mechanisms and management of RSA are mired by substandard design and lack of optimal randomized clinical trials and have resulted in disagreement on guidelines for evaluation and treatments for patients with multiple pregnancy losses of unknown etiology. The present review focuses on evidence-based research discussion with immunologic causes, and immune-regulatory therapies recommended for helping patients with a history of RSA. We highlight data that might support revalidation of low molecular weight heparin as a protective therapy in RSA. Newly launched growth factors, GM-CSF, and potentially novel agents to suppress inflammatory rejection, including regulatory T cells, human chorionic gonadotropin, and M-CSF/IL-10, may work in concert with tender-loving-care therapy and give hope to couples with multiple pregnancy losses.
Assuntos
Aborto Habitual/imunologia , Aborto Habitual/terapia , Gonadotropina Coriônica/uso terapêutico , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/uso terapêutico , Heparina de Baixo Peso Molecular/uso terapêutico , Humanos , Recém-Nascido , Interleucina-10/uso terapêutico , Fator Estimulador de Colônias de Macrófagos/uso terapêutico , Gravidez , Resultado da Gravidez , Linfócitos T Reguladores/imunologiaRESUMO
Type 5 acid phosphatase (ACP5; also known as tartrate-resistant acid phosphatase or uteroferrin) is a metalloprotein secreted by the endometrial glandular epithelium of pigs, mares, sheep, and water buffalo. In this paper, we describe the phylogenetic distribution of endometrial expression of ACP5 and demonstrate that endometrial expression arose early in evolution (i.e., before divergence of prototherian and therian mammals ~166 million years ago). To determine expression of ACP5 in the pregnant endometrium, RNA was isolated from rhesus, mouse, rat, dog, sheep, cow, horse, armadillo, opossum, and duck-billed platypus. Results from RT-PCR and RNA-Seq experiments confirmed that ACP5 is expressed in all species examined. ACP5 was also demonstrated immunochemically in endometrium of rhesus, marmoset, sheep, cow, goat, and opossum. Alignment of inferred amino acid sequences shows a high conservation of ACP5 throughout speciation, with species-specific differences most extensive in the N-terminal and C-terminal regions of the protein. Analysis by Selecton indicated that most of the sites in ACP5 are undergoing purifying selection, and no sites undergoing positive selection were found. In conclusion, endometrial expression of ACP5 is a common feature in all orders of mammals and has been subjected to purifying selection. Expression of ACP5 in the uterus predates the divergence of therians and prototherians. ACP5 is an evolutionary conserved gene that likely exerts a common function important for pregnancy in mammals using a wide range of reproductive strategies.
Assuntos
Fosfatase Ácida/metabolismo , Evolução Biológica , Endométrio/metabolismo , Isoenzimas/metabolismo , Mamíferos/genética , Prenhez/genética , Fosfatase Ácida/genética , Sequência de Aminoácidos , Animais , Feminino , Isoenzimas/genética , Mamíferos/fisiologia , Dados de Sequência Molecular , Filogenia , Gravidez , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Fosfatase Ácida Resistente a TartaratoRESUMO
The 31st Annual Meeting of the American Society for Reproductive Immunology provided an excellent platform for basic and clinical scientists to brainstorm on current reproductive health issues such as repeated implantation and pregnancy failure, preterm birth, preeclampsia and genital tract infections such as HIV. The goal of the meeting was to foster cross-pollination of ideas as well as to encourage participation of young investigators in the field. The conference was preceded by the 4th Annual Post-Graduate Workshop with the theme of bringing bedside problems to the bench and facilitating collaboration between clinicians and basic scientists. Christopher Davies and Richard Bronson chaired the conference, which hosted approximately 180 delegates representing more than 26 countries across Asia, Australia, Latin America, Europe and North America.
Assuntos
Congressos como Assunto , Complicações na Gravidez/imunologia , Complicações na Gravidez/metabolismo , Reprodução/imunologia , Sociedades Médicas , Animais , Feminino , Humanos , Gravidez , Complicações na Gravidez/epidemiologia , Complicações na Gravidez/terapia , Estados UnidosRESUMO
PROBLEM: Regulatory T cells (T(reg) ) are a vital immune cellular population at the maternal-fetal interface. They are likely to aid in immune tolerance by dampening the harmful effects of other immune cellular populations through cell-cell-mediated interactions as well as by producing IL-10 and TGF-ß. In addition to the anti-inflammatory properties, IL-10 has emerged as an important vascular cytokine choreographing endovascular interactions and angiogenesis and regulates hypertension. METHOD OF STUDY: Review of innovative concepts to understand the temporal role of T(regs) in both mouse and human pregnancy, particularly whether uterine T(reg) play a potential role in regulating vascular homeostasis and blood flow during pregnancy. RESULTS: T(reg) guard immune tolerance, getting cytotoxically activated under certain conditions, leading to adverse pregnancy outcome. CONCLUSION: Despite increasing evidence of T(reg) tissue-specific expansion and functional plasticity, their role in vascular activity, pre-eclampsia or gestational diabetes is obscure and needs closer investigation to delineate its role later during pregnancy.
Assuntos
Hipertensão/imunologia , Interleucina-10/imunologia , Complicações Cardiovasculares na Gravidez/imunologia , Linfócitos T Reguladores/imunologia , Útero/imunologia , Animais , Feminino , Humanos , Hipertensão/metabolismo , Interleucina-10/metabolismo , Gravidez , Complicações Cardiovasculares na Gravidez/metabolismo , Útero/metabolismoRESUMO
IL-10 is a pregnancy compatible cytokine that plays a vital role in maintaining the balance of anti-inflammatory and pro-inflammatory milieu at the maternal-fetal interface. Recent evidence now suggests that IL-10 is a potent vascular cytokine that can blunt hypertension and inflammation-mediated vascular dysfunction. Thus, a re-evaluation of IL-10 as a cytokine supporting endovascular interactions and angiogenesis as well as blunting hypoxic-injury and preeclampsia-like features is warranted. In this review, we highlight these novel functions of IL-10 and propose that its immune-modulating and vascular functions are mutually inclusive, particularly in the context of normal gestation.
Assuntos
Células Endoteliais/imunologia , Imunoterapia , Interleucina-10/imunologia , Placenta/imunologia , Pré-Eclâmpsia/imunologia , Animais , Citoproteção , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Feminino , Humanos , Hipertensão , Hipóxia , Tolerância Imunológica , Imunoterapia/tendências , Inflamação , Interleucina-10/uso terapêutico , Neovascularização Fisiológica/imunologia , Placenta/irrigação sanguínea , Placenta/patologia , Placentação , Pré-Eclâmpsia/fisiopatologia , Pré-Eclâmpsia/terapia , GravidezRESUMO
Hypoxia has been implicated in the pathogenesis of preeclampsia, a hypertensive disorder of pregnancy. However, in vivo evidence and mechanistic understanding remain elusive. Preeclampsia is associated with impaired placental angiogenesis. We have recently shown that interleukin (IL)-10 can support trophoblast-driven endovascular crosstalk. Accordingly, we hypothesize that pathological levels of oxygen coupled with IL-10 deficiency induce severe preeclampsia-like features coupled with elevated production of antiangiogenic factors, apoptotic pathways, and placental injury. Exposure of pregnant wild-type and IL-10(-/-) mice to 9.5% oxygen resulted in graded placental injury and systemic symptoms of renal pathology, proteinuria (wild-type 645.15 ± 115.73 versus 198.09 ± 93.45; IL-10(-/-) 819.31 ± 127.85 versus 221.45 ± 82.73 µg/mg/24 hours) and hypertension (wild-type 118.37 ± 14.45 versus 78.67 ± 14.07; IL-10(-/-) 136.03 ± 22.59 versus 83.97 ± 18.25 mm Hg). Recombinant IL-10 reversed hypoxia-induced features in pregnant IL-10(-/-) mice confirming the protective role of IL-10 in preeclampsia. Hypoxic exposure caused marked elevation of soluble fms-like tyrosine kinase 1 (110.8 ± 20.1 versus 44.7 ± 11.9 ng/mL) in IL-10(-/-) mice compared with their wild-type counterparts (81.6 ± 13.1 versus 41.2 ± 8.9 ng/mL), whereas soluble endoglin was induced to similar levels in both strains (approximately 380 ± 50 versus 180 ± 31 ng/mL). Hypoxia-induced elevation of p53 was associated with marked induction of proapoptotic protein Bax, downregulation of Bcl-2, and trophoblast-specific apoptosis in utero-placental tissue. Collectively, we conclude that severe preeclampsia pathology could be triggered under certain threshold oxygen levels coupled with intrinsic IL-10 deficiency, which lead to excessive activation of antiangiogenic and apoptotic pathways.
Assuntos
Hipóxia/complicações , Interleucina-10/metabolismo , Placenta/fisiopatologia , Pré-Eclâmpsia/etiologia , Animais , Apoptose/fisiologia , Modelos Animais de Doenças , Feminino , Hipóxia/metabolismo , Hipóxia/fisiopatologia , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Knockout , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Pré-Eclâmpsia/fisiopatologia , Gravidez , Proteinúria/etiologia , Proteinúria/metabolismo , Proteinúria/fisiopatologia , Transdução de Sinais/fisiologia , Trofoblastos/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Early diagnosis and treatment of preeclampsia would significantly reduce maternal and fetal morbidity and mortality. However, its etiology and prediction have remained elusive. Based on the hypothesis that sera from patients with preeclampsia could function as a "blueprint" of causative factors, we describe a serum-based pregnancy-specific mouse model that closely mirrors the human condition as well as an in vitro predictive assay. We show that a single administration of human preeclampsia serum in pregnant IL-10-/- mice induced the full spectrum of preeclampsia-like symptoms, caused hypoxic injury in uteroplacental tissues, and elevated soluble fms-like tyrosine kinase 1 and soluble endoglin, markers thought to be related to the disease. The same serum sample(s) induced a partial preeclampsia phenotype in wild-type mice. Importantly, preeclampsia serum disrupted cross talk between trophoblasts and endothelial cells in an in vitro model of endovascular activity. Disruption of endovascular activity could be documented in serum samples as early as 12 to 14 weeks of gestation from patients who subsequently developed preeclampsia. These results indicate that preeclampsia patient sera can be used to understand the pregnancy-specific disease pathology in mice and can predict the disorder.
Assuntos
Bioensaio/métodos , Modelos Animais de Doenças , Pré-Eclâmpsia/sangue , Pré-Eclâmpsia/diagnóstico , Gravidez/sangue , Soro , Animais , Pressão Sanguínea , Feminino , Idade Gestacional , Humanos , Hipóxia , Interleucina-10/sangue , Interleucina-10/genética , Interleucina-10/imunologia , Rim/patologia , Camundongos , Camundongos Knockout , Pré-Eclâmpsia/imunologia , Gravidez/imunologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/sangueRESUMO
Taking a cue from the recent workshop 'Preeclampsia--a Pressing Problem' sponsored by the National Institutes of Child Health and Human Development, this review article takes a fresh look at hypoxia and a dysfunctional immune system as the key contributors to the etiology of preeclampsia and the mechanisms involved therein. In the context of epidemiological research on the intricate and multifactorial nature of preeclampsia, we focus on hypoxia as an upstream regulator of preeclampsia and its consequences in a model compromised by a deficiency in key pregnancy compatible immune modulators. It has been proposed that placental hypoxia releases cytotoxic factors produced at the maternal-fetal interface into the circulation to manifest the maternal symptoms associated with preeclampsia. However, it is not clear how this mechanism is empowered in pregnant women. Does systemic hypoxia exert preeclampsia-like effects on pregnancy? Are these effects further manifested by intrinsic inflammation in the absence of key immune modulators such as IL-10? Thus, it is of paramount importance that in vivo models be developed wherein the role of systemic hypoxia can be evaluated for preeclampsia-causing events. We present a discussion on whether prolonged exposure to hypoxia can lead to a perpetual cycle of compartmentalized uteroplacental tissue damage, release of anti-angiogenic and vasoconstrictive factors that impair trophoblast invasion and promote systemic vascular resistance resulting in the maternal syndrome.
Assuntos
Hipóxia Celular/imunologia , Pré-Eclâmpsia/etiologia , Animais , Feminino , Humanos , Troca Materno-Fetal/imunologia , Placenta/irrigação sanguínea , Placenta/fisiopatologia , GravidezRESUMO
Despite intense investigation, preeclampsia (PE) remains largely enigmatic. Relatively late onset of diagnostic signs and heterogeneous nature of the disease further contribute to poor understanding of its etiology and clinical management. There exist no concrete animal models that can provide mechanistic underpinnings for evaluating targeted therapeutic intervention. Poor cross-sectional findings with potential biochemical markers reported so far have proved counterintuitive and suggest a need for novel approaches to predict the early onset of disease. Because of the co-onset of local placental anomalies and systemic manifestation of symptoms, it is highly likely that serum from PE patients can provide a "blueprint" of causative factors. Proteomic and/or functional analysis of maternal serum are expected to predict the onset of disease ahead of manifestation of clinical symptoms. A serum-based predictive assay should overcome complexities resulting from the heterogeneous etiology of PE. This review attempts to address some of these issues and discuss the signature biochemical serum factors and propose new and better ways to predict PE.
Assuntos
Biomarcadores/sangue , Pré-Eclâmpsia , Testes Sorológicos , Animais , Feminino , Humanos , Placentação , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/etiologia , Pré-Eclâmpsia/patologia , Gravidez , Prognóstico , ProteômicaRESUMO
Despite serious health risks in humans and wild life, the underlying mechanisms that explain the gene-environment effects of chemical toxicants are largely unknown. Polychlorinated biphenyls (PCBs) are one of the most ubiquitous environmental toxicants worldwide, with reported epidemiological evidence for reproductive and neurocognitive anomalies in humans. Here, we show that Aroclor 1254, a mixture of structurally distinct PCBs, causes preterm birth in interleukin (IL)-10(-/-) mice at a dose that does not show any adverse effects in wild type mice, highlighting the significance of IL-10 as an anti-toxicant cytokine. Aroclor 1254-treated IL-10(-/-) mice demonstrated increased amniotic fluid, intrauterine growth restriction, and reduced litter size with postnatal neuromotor defects. Further, our results identify aquaporin 1 (AQP1), a potent effector of fluid volume regulation and angiogenic activity, as a novel placental target of PCBs. In vivo or in vitro exposure to Aroclor 1254 coupled with IL-10 deficiency significantly reduced the protein content of AQP1. Reduced uterine AQP1 levels were associated with defective spiral artery transformation. Importantly, recombinant IL-10 reversed PCB-induced in vivo and in vitro effects. These data demonstrate for the first time that the IL-10-AQP1 axis is a novel regulator of PCB-induced in utero effects.
Assuntos
Aquaporina 1/metabolismo , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Animais , Anticorpos Bloqueadores/farmacologia , Aquaporina 1/antagonistas & inibidores , Artérias/efeitos dos fármacos , Transporte Biológico/efeitos dos fármacos , Comunicação Celular/efeitos dos fármacos , Linhagem Celular , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Feminino , Humanos , Interleucina-10/deficiência , Camundongos , Camundongos Endogâmicos C57BL , Doenças do Sistema Nervoso/induzido quimicamente , Doenças do Sistema Nervoso/patologia , Placenta/efeitos dos fármacos , Placenta/metabolismo , Gravidez , Primeiro Trimestre da Gravidez/efeitos dos fármacos , Nascimento Prematuro/induzido quimicamente , Nascimento Prematuro/metabolismo , Trofoblastos/citologia , Trofoblastos/efeitos dos fármacosRESUMO
Neuroblastoma is the most common extracranial solid tumor in children and, when disseminated, carries a poor prognosis. Even with aggressive combinations of chemotherapy, surgery, autologous bone marrow transplant, and radiation, long-term survival remains at 30% and new therapies are needed. Recently, a patient with neuroblastoma who acquired Chagas disease was treated with nifurtimox with subsequent reduction in tumor size. The effect of nifurtimox on the neuroblastoma cell lines CHLA-90, LA1-55n, LA-N2, SMS-KCNR, and SY5Y was examined. Nifurtimox decreased cell viability in a concentration-dependent manner. Cell morphology, terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay, and caspase-3 activation indicate that cell death was primarily due to apoptosis. Nifurtimox also suppressed basal and TrkB-mediated Akt phosphorylation, and the cytotoxicity of nifurtimox was attenuated by a tyrosine hydroxylase inhibitor (alpha-methyl-tyrosine). Nifurtimox killed catecholaminergic, but not cholinergic, autonomic neurons in culture. In vivo xenograft models showed inhibition of tumor growth with a histologic decrease in proliferation and increase in apoptosis. These results suggest that nifurtimox induces cell death in neuroblastoma. Therefore, further studies are warranted to develop nifurtimox as a promising new treatment for neuroblastoma.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neuroblastoma/tratamento farmacológico , Nifurtimox/farmacologia , Animais , Western Blotting , Caspase 3/efeitos dos fármacos , Catecolaminas/metabolismo , Linhagem Celular Tumoral , Fragmentação do DNA/efeitos dos fármacos , Feminino , Humanos , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Nus , Neurônios/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Espécies Reativas de Oxigênio , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Although replete with cytotoxic machinery, uterine NK (uNK) cells remain tolerant at the maternal-fetal interface. The mechanisms that facilitate the uNK cell tolerance are largely unknown. In this study, we demonstrate that vascular endothelial growth factor (VEGF) C, a proangiogenic factor produced by uNK cells, is responsible for their noncytotoxic activity. VEGF C-producing uNK cells support endovascular processes as demonstrated in a three-dimensional coculture model of capillary tube formation on Matrigel. Peripheral blood NK cells fail to produce VEGF C and remain cytotoxic. This response can be reversed by exogenous VEGF C. We show that cytoprotection by VEGF C can be related to induction of the TAP-1 expression and MHC class I assembly in target cells. Small interfering RNA-mediated silencing of TAP-1 expression abolished the VEGF C-imparted protection. Overall, these results demonstrate that empowerment of uNK cells with angiogenic factors keeps them noncytotoxic. This phenotype is critical to their pregnancy-compatible immunovascular role during placentation and fetal development.
Assuntos
Decídua/imunologia , Tolerância Imunológica/imunologia , Células Matadoras Naturais/imunologia , Gravidez/imunologia , Útero/imunologia , Fator C de Crescimento do Endotélio Vascular/imunologia , Adulto , Western Blotting , Técnicas de Cocultura , Citotoxicidade Imunológica , Decídua/citologia , Feminino , Feto/imunologia , Citometria de Fluxo , Humanos , Neovascularização Fisiológica/imunologia , Fator C de Crescimento do Endotélio Vascular/metabolismoRESUMO
The immune tolerance and de novo vascularization are two highly intriguing processes at the maternal-fetal interface that appear to be central to normal pregnancy outcome. Immune tolerance occurs despite the local presence of an active maternal immune system including macrophages, dendritic cells and specialized CD56(bright)CD16(-) uterine natural killer (uNK) cells (65-70%). Recent observations indicate that the phenotypic and functional repertoire of uNK cells is distinct from peripheral blood NK and endometrial NK cells, challenging the understanding of their temporal occurrence and function. Origin and specialized programming of uNK cells continue to be debated. uNK cells, replete with an armamentarium to kill the foreign, tolerate the conceptus and facilitate pregnancy. Why do these uNK cells remain non-cytotoxic? Are these NK cells 'multitasking' in nature harboring beneficial and detrimental roles in pregnancy? Are there distinct subpopulations of NK cells that may populate the decidua? We propose that the endometrium/decidua functions as an 'inducible tertiary lymphoid tissue' that supports the recruitment and expansion of CD56(bright)CD16(-) NK cells and induces transcriptional up-regulation of angiogenic machinery in response to exposure to local hormonal factors, cytokine milieu and perhaps hypoxia. The angiogenic features of uNK cells could further result in a 'multitasking' phenotype that still remains to be characterized. This article discusses the factors and pathways that bridge the angiogenic and non-cytotoxic response machineries at the maternal-fetal interface.
Assuntos
Tolerância Imunológica , Células Matadoras Naturais/imunologia , Subpopulações de Linfócitos/imunologia , Útero/imunologia , Animais , Linhagem da Célula , Feminino , Histocompatibilidade Materno-Fetal/imunologia , Humanos , Troca Materno-Fetal/imunologia , Neovascularização Fisiológica/imunologia , Fenótipo , Gravidez , Complicações na Gravidez/imunologia , Útero/irrigação sanguíneaRESUMO
The cytotoxic, anti-proliferative and apoptotic effects of 3-Bromoacetoxy Calcidiol (B3CD), a derivative of vitamin D3 precursor calcidiol, on human neuroblastoma (NB) cells were examined. NB, predominantly a tumor of early childhood, is the most common extracranial solid tumor. Despite aggressive treatments, survival for advanced stages remains low and novel treatment strategies are needed. B3CD-induced apoptosis in various neuroblastic cells via caspases-3 and -9 activation. B3CD upregulated mitochondrial pro-apoptotic Bax and anti-apoptotic Bcl-2 expression, caused cytochrome c release, downregulated N-Myc expression and activated pro-survival marker Akt. Accordingly, B3CD treatment dose dependently reduced the viability of NB cells with IC50 values between 1 and 3 microm. The cytotoxicity of B3CD was significantly higher than for the calcemic parent-compound vitamin D3 (IC50 between 10 and 30 microm). Further studies revealed that B3CD treatment inhibits the proliferation of NB cells at low concentrations (IC50 between 30 and 100 nm). Cell cycle analysis showed a dramatic increase in the apoptotic sub-diploidal population along with a cell cycle block. In summary, the present study shows that B3CD is toxic to NB cells via suppression of cell proliferation and cell viability by caspase activation and regulation of survival signals. These results suggest that B3CD could be developed as a treatment for NB.
Assuntos
Apoptose/efeitos dos fármacos , Compostos de Bromo , Calcifediol/análogos & derivados , Proliferação de Células , Biomarcadores/metabolismo , Conservadores da Densidade Óssea/química , Conservadores da Densidade Óssea/farmacologia , Conservadores da Densidade Óssea/uso terapêutico , Compostos de Bromo/química , Compostos de Bromo/farmacologia , Compostos de Bromo/uso terapêutico , Calcifediol/química , Calcifediol/farmacologia , Calcifediol/uso terapêutico , Calcitriol/metabolismo , Calcitriol/farmacologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular , Humanos , Estrutura Molecular , Neuroblastoma/tratamento farmacológico , Neuroblastoma/patologiaRESUMO
Lipophilic extracts of Ginkgo biloba L. leaves were tested for their possible role on rodent models of depression and stress. Lipophilic extracts of Ginkgo leaves (LEG) at (50 and 100 mg/kg, p.o.) exhibited dose dependent, significant antidepressant activity in the behavioral despair test and learned helplessness rodent model of depression. The activities were comparable to that of imipramine (15 mg/kg) and EGb 761 (50 mg/kg). In the cold immobilization stress induced gastric ulcer model of stress, only the LEG showed a significant reduction in the ulcer index. GC-MS characterization of this bioactive extract was found to be rich in a group of 6-alkyl salicylates (6-AS), along with a fatty alcohol, fatty acids and cardanols. The n-heptadecenyl salicylate represented 60% of the 6-AS. Notable was the absence of dihydroxy alkylphenols which are linked to allergic reactions similar to the urushiols present in poison ivy. In commercial products of Ginkgo, these dihydroxy phenols as well as the favorable 6-AS are removed during enrichment of flavonol glycosides and terpenic lactones. The current findings suggest that intact carboxylic acid groups containing 6-AS are the bioactive components of the lipophilic extract of Ginkgo leaves with antidepressant and antistress activities.
Assuntos
Antidepressivos/análise , Ginkgo biloba/química , Fitoterapia , Extratos Vegetais/uso terapêutico , Estresse Fisiológico/tratamento farmacológico , Estresse Psicológico/tratamento farmacológico , Animais , Cromatografia Gasosa-Espectrometria de Massas , Extratos Vegetais/química , Folhas de Planta/química , RatosRESUMO
Advanced ovarian cancer (OC) is not curable by surgery alone and chemotherapy is essential for its treatment. Isothiocyanates have been shown to inhibit carcinogen-induced tumorigenesis in animal models, yet no efforts have been made to determine their therapeutic potential in OC. In the present study, we investigated the mechanism of the anti-proliferative and apoptotic activity of benzyl isothiocyanate (BITC) in OC. BITC inhibited the proliferation of OC cells and induced apoptosis in OC cells. Apoptosis was induced by a strong activation of caspase-3 and -9, and cleavage of PARP-1. However, caspase-8 was not activated by BITC. Cytotoxic effects of BITC were reversed by the inhibition caspase-3 and -9 specific inhibitors. BITC showed a concentration dependent decrease in the levels of Bcl-2 with a concomitant increase in Bax levels. In addition, BITC activated proapoptotic signaling by phosphorylation JNK1/2 and p38 while simultaneously inhibiting survival signaling mediated by ERK1/2 and Akt phosphorylation in a dose-dependent manner. While JNK inhibitor SP600125 and p38 inhibitor SB203580, abolished the cytotoxic effect of BITC, MEK inhibitor, PD98059 and PI3 kinase inhibitor, LY294002 failed to show such reversal indicating a critical role played by JNK1/2 and p38 signaling in apoptosis induced by BITC. In summary, our studies demonstrate that BITC inhibits proliferation of OC cells and induces apoptosis via caspase-9 and -3 pathways. BITC inhibits ERK1/2 and Akt survival signaling while simultaneously activating pro-apoptotic p38 and JNK1/2. Therefore, BITC can be potentially developed as a therapeutic agent to treat OC.
Assuntos
Antineoplásicos/farmacologia , Apoptose , Isotiocianatos/farmacologia , Neoplasias Ovarianas/tratamento farmacológico , Animais , Caspase 3/metabolismo , Caspase 9/metabolismo , Replicação do DNA/efeitos dos fármacos , Ativação Enzimática , Inibidores Enzimáticos/farmacologia , Feminino , Células HL-60 , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RatosRESUMO
BACKGROUND: Chemotherapy-resistant neuroblastoma is a difficult disease to treat with poor survival. OBSERVATIONS: We treated a patient with neuroblastoma who had progressed on conventional chemotherapy. This 5-year-old girl with chemotherapy-resistant neuroblastoma developed Chagas disease at the start of salvage chemotherapy for which she was also started on nifurtimox. The neuroblastoma response to these treatments resulted in clinical remission. In vitro, treatment of a neuroblastoma cell line with nifurtimox resulted in decreased cell viability whereas no effect was seen on an endothelial cell line. CONCLUSIONS: Nifurtimox shows promise as a potential new treatment for neuroblastoma and warrants further testing.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Doença de Chagas/tratamento farmacológico , Neuroblastoma/tratamento farmacológico , Nifurtimox/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Doença de Chagas/complicações , Doença de Chagas/diagnóstico , Pré-Escolar , Ciclofosfamida/administração & dosagem , Progressão da Doença , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Neuroblastoma/complicações , Neuroblastoma/diagnóstico , Nifurtimox/administração & dosagem , Nifurtimox/farmacologia , Recidiva , Indução de Remissão , Tomografia Computadorizada por Raios X/métodos , Topotecan/administração & dosagem , Resultado do TratamentoRESUMO
Angiogenesis is a complex process involving coordinated steps of endothelial cell activation, proliferation, migration, tube formation and capillary sprouting with participation of intracellular signaling pathways. Regulation of angiogenesis carries tremendous potential for cancer therapy. Our earlier studies showed that vitamin D-binding protein-macrophage activating factor (DBP-maf) acts as a potent anti-angiogenic factor and inhibits tumor growth in vivo. The goal of this investigation was to understand the effect of DBP-maf on human endothelial cell (HEC) and the mechanism of angiogenesis inhibition. DBP-maf inhibited human endothelial cell (HEC) proliferation by inhibiting DNA synthesis (IC(50) = 7.8 +/- 0.15 microg/ml). DBP-maf significantly induced S- and G0/G1-phase arrest in HEC in 72 h. DBP-maf potently blocked VEGF-induced migration, tube-formation of HEC in a dose dependent manner. In addition, DBP-maf inhibited growth factor-induced microvessel sprouting in rat aortic ring assay. Moreover, DBP-maf inhibited VEGF signaling by decreasing VEGF-mediated phosphorylation of VEGFR-2 and ERK1/2, a downstream target of VEGF signaling cascade. However, Akt activation was not affected. These studies collectively demonstrate that DBP-maf inhibits angiogenesis by blocking critical steps such as HEC proliferation, migration, tube formation and microvessel sprouting. DBP-maf exerts its effect by inhibiting VEGR-2 and ERK1/2 signaling cascades. Understanding the cellular and molecular mechanisms of anti-endothelial activity of DBP-maf will allow us to develop it as an angiogenesis targeting novel drug for tumor therapy.