RESUMEN
AIMS: Differentiation-inducing factor-1 (DIF-1) is a polyketide produced by Dictyostelium discoideum that inhibits growth and migration, while promoting the differentiation of Dictyostelium stalk cells through unknown mechanisms. DIF-1 localizes in stalk mitochondria. In addition to its effect on Dictyostelium, DIF-1 also inhibits growth and migration, and induces mitochondrial fission followed by mitophagy in mammalian cells, at least in part by activating AMP-activated protein kinase (AMPK). In a previous study, we found that DIF-1 binds to mitochondrial malate dehydrogenase (MDH2) and inhibits its activity in HeLa cells. In the present study, we investigated whether MDH2 serves as a pharmacological target of DIF-1 in mammalian cells. MAIN METHODS: To examine the enzymatic activity of MDH, mitochondrial morphology, and molecular mechanisms of DIF-1 action, we conducted an MDH reverse reaction assay, immunofluorescence staining, western blotting, and RNA interference using mammalian cells such as human umbilical vein endothelial cells, human cervical cancer cells, mouse endothelial cells, and mouse breast cancer cells. KEY FINDINGS: DIF-1 inhibited mitochondrial but not cytoplasmic MDH activity. Similar to DIF-1, LW6, an authentic MDH2 inhibitor, induced phosphorylation of AMPK, resulting in the phosphorylation of acetyl-CoA carboxylase (ACC) and the dephosphorylation of p70 S6 kinase with approximately the same potency. DIF-1 and LW6 induced mitochondrial fission. Furthermore, MDH2 knockdown using siRNA reproduced the DIF-1 action on the AMPK signaling and mitochondrial morphology. Conversely, an AMPK inhibitor prevented DIF-1-induced mitochondrial fission. SIGNIFICANCE: We propose that MDH2 is a mammalian target of DIF-1 for the activation of AMPK and induction of mitochondrial fission.
Asunto(s)
Proteínas Quinasas Activadas por AMP , Malato Deshidrogenasa , Mitocondrias , Dinámicas Mitocondriales , Humanos , Proteínas Quinasas Activadas por AMP/metabolismo , Dinámicas Mitocondriales/efectos de los fármacos , Dinámicas Mitocondriales/fisiología , Malato Deshidrogenasa/metabolismo , Mitocondrias/metabolismo , Células HeLa , Animales , Hexanonas/farmacología , Hexanonas/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Activación Enzimática , Hidrocarburos CloradosRESUMEN
The spread of malarial parasites resistant to first-line treatments such as artemisinin combination therapies is a global health concern. Differentiation-inducing factor 1 (DIF-1) is a chlorinated alkylphenone (1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl) hexan-1-one) originally found in the cellular slime mould Dictyostelium discoideum. We previously showed that some derivatives of DIF-1, particularly DIF-1(+2) (1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl) octan-1-one), exert potent antimalarial activities. In this study, we synthesised DIF-1(+3) (1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl) nonan-1-one). We then evaluated the effects of DIF-1(+3) in vitro on Plasmodium falciparum and in vivo over 7 days (50-100 mg/kg/day) in a mouse model of Plasmodium berghei. DIF-1(+3) exhibited a half-maximal inhibitory concentration of approximately 20-30 % of DIF-1(+2) in three laboratory strains with a selectivity index > 263, including in strains resistant to chloroquine and artemisinin. Parasite growth and multiplication were almost completely suppressed by treatment with 100 mg/kg DIF-1(+3). The survival time of infected mice was significantly increased (P = 0.006) with no apparent adverse effects. In summary, addition of an acyl group to DIF-1(+2) to prepare DIF-1(+3) substantially enhanced antimalarial activity, even in drug-resistant malaria, indicating the potential of applying DIF-1(+3) for malaria treatment.
Asunto(s)
Antimaláricos , Hexanonas , Plasmodium falciparum , Antimaláricos/farmacología , Animales , Ratones , Hexanonas/farmacología , Hexanonas/química , Plasmodium falciparum/efectos de los fármacos , Plasmodium berghei/efectos de los fármacos , Malaria/tratamiento farmacológico , Malaria/parasitología , Dictyostelium/efectos de los fármacos , Acilación , Femenino , Hidrocarburos CloradosRESUMEN
3-Hydroxyhexan-2-one (3-C6-ketol) has emerged as the most conserved pheromone structure within the beetle family Cerambycidae. In this study, we report the sex-specific production of this compound by males of 12 species of South American cerambycid beetles. Males of Chrysoprasis chalybea Redtenbacher and Mallosoma zonatum (Sahlberg) (Tribe Dichophyiini), and Ambonus lippus (Germar), Eurysthea hirta (Kirby), Pantonyssus nigriceps Bates, Stizocera plicicollis (Germar), and Stizocera tristis (Guérin-Méneville) (Elaphidiini) produced 3R-C6-ketol as a single component, whereas males of Neoclytus pusillus (Laporte & Gory) (Clytini), Aglaoschema concolor (Gounelle), Orthostoma abdominale (Gyllenhal) (Compsocerini), Dorcacerus barbatus (Olivier), and Retrachydes thoracicus thoracicus (Olivier) (Trachyderini) produced 3R-C6-ketol, along with lesser amounts of other compounds. In field trials testing 8 known cerambycid pheromone compounds, C. chalybea, E. hirta, and R. t. thoracicus were attracted in significant numbers to traps baited with 3-C6-ketol. A second field experiment provided support for the strategy of using the attraction of cerambycid species to test lures as a method of providing leads to their likely pheromone components. Because both sexes are attracted to these aggregation-sex pheromones, live beetles can be obtained from baited traps to verify they produce the compound(s) to which they were attracted, that is, that the compounds are indeed pheromone components.
Asunto(s)
Escarabajos , Animales , Masculino , Femenino , Hexanonas/farmacología , Feromonas/farmacología , Atractivos Sexuales/farmacología , Especificidad de la Especie , América del SurRESUMEN
Differentiation-inducing factor 1 (DIF-1) is a morphogen produced by Dictyostelium discoideum that inhibits the proliferation and migration of both D. discoideum and most mammalian cells. Herein, we assessed the effect of DIF-1 on mitochondria, because DIF-3, which is similar to DIF-1, reportedly localizes in the mitochondria when added exogenously, however the significance of this localization remains unclear. Cofilin is an actin depolymerization factor that is activated by dephosphorylation at Ser-3. By regulating the actin cytoskeleton, cofilin induces mitochondrial fission, the first step in mitophagy. Here, we report that DIF-1 activates cofilin and induces mitochondrial fission and mitophagy mainly using human umbilical vein endothelial cells (HUVECs). AMP-activated kinase (AMPK), a downstream molecule of DIF-1 signaling, is required for cofilin activation. Pyridoxal phosphatase (PDXP)-known to directly dephosphorylate cofilin-is also required for the effect of DIF-1 on cofilin, indicating that DIF-1 activates cofilin through AMPK and PDXP. Cofilin knockdown inhibits mitochondrial fission and decreases mitofusin 2 (Mfn2) protein levels, a hallmark of mitophagy. Taken together, these results indicate that cofilin is required for DIF-1- induced mitochondrial fission and mitophagy.
Asunto(s)
Dictyostelium , Hexanonas , Animales , Humanos , Proteínas Quinasas Activadas por AMP , Factores Despolimerizantes de la Actina/metabolismo , Factores Despolimerizantes de la Actina/farmacología , Dinámicas Mitocondriales , Dictyostelium/metabolismo , Células Endoteliales/metabolismo , Diferenciación Celular , Monoéster Fosfórico Hidrolasas , Piridoxal/farmacología , Hexanonas/farmacología , Mamíferos/metabolismoRESUMEN
Malaria, which is caused by protozoa of the genus Plasmodium, remains a major endemic public health problem worldwide. Since artemisinin combination therapies are used as a first-line treatment in all endemic regions, the emergence of parasites resistant to these regimens has become a serious problem. Differentiation-inducing factor 1 (DIF-1) is a chlorinated alkylphenone originally found in the cellular slime mold Dictyostelium discoideum. DIF-1 and its derivatives exhibit a range of biological activities. In the present study, we investigated the effects of 41 DIF derivatives on the growth of Plasmodium falciparum in vitro using four laboratory strains and 12 field isolates. Micromolar concentrations of several DIF derivatives strongly suppressed the growth of the four laboratory strains, including strains that exhibited resistance to chloroquine and artemisinin, as well as strains that were susceptible to these drugs. In addition, DIF-1(+2), the most potent derivative, strongly suppressed the growth of 12 field isolates. We also examined the effects of DIF-1(+2) on the activity of the rodent malarial parasite Plasmodium berghei in mice. Intraperitoneal administration of DIF-1(+2) over 4 days (50 or 70 mg/kg/day) significantly suppressed the growth of the parasite in the blood with no apparent adverse effects, and a dose of 70 mg/kg/day significantly prolonged animal survival. These results suggest that DIF derivatives, such as DIF-1(+2), could serve as new lead compounds for the development of antimalarial agents.
Asunto(s)
Antimaláricos/farmacología , Dictyostelium , Hexanonas/farmacología , Parásitos/crecimiento & desarrollo , Plasmodium berghei/crecimiento & desarrollo , Plasmodium falciparum/crecimiento & desarrollo , Células 3T3-L1 , Animales , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Parásitos/efectos de los fármacos , Plasmodium berghei/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacosRESUMEN
Here, we study the pheromone chemistry of two South American cerambycid beetle species, and their behavioral responses to candidate pheromone components. Adult males of Stizocera phtisica Gounelle (subfamily Cerambycinae: tribe Elaphidiini) produced a sex-specific blend of (R)-3-hydroxyhexan-2-one with lesser amounts of 3-methylthiopropan-1-ol. In field bioassays, traps baited with racemic 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol did not catch conspecific beetles, but did catch both sexes of a sympatric species, Chydarteres dimidiatus dimidiatus (F.) (Cerambycinae: Trachyderini). We found that males of this species also produce (R)-3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol, and small amounts of 2-phenylethanol. Subsequent bioassays with these compounds showed that a blend of 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol constitutes the aggregation-sex pheromone of C. d. dimidiatus, with 2-phenylethanol not influencing the attraction of conspecifics. During the field bioassays, six other species in the Cerambycinae also were caught in significant numbers, including Aglaoschema ventrale (Germar) (tribe Compsocerini), congeners Chrysoprasis aurigena (Germar), Chrysoprasis linearis Bates, and an unidentified Chrysoprasis species (Dichophyiini), and Cotyclytus curvatus (Germar) and Itaclytus olivaceus (Laporte & Gory) (both Clytini), suggesting that one or more of the compounds tested are also pheromone components for these species.
Asunto(s)
Escarabajos/efectos de los fármacos , Feromonas/farmacología , Animales , Escarabajos/fisiología , Hexanonas/farmacología , Masculino , América del Sur , Especificidad de la EspecieRESUMEN
Increasing evidence suggests that n-hexane induces nerve injury via neuronal apoptosis induced by its active metabolite 2,5-hexanedione (HD). However, the underlying mechanism remains unknown. Studies have confirmed that pro-nerve growth factor (proNGF), a precursor of mature nerve growth factor (mNGF), might activate apoptotic signaling by binding to p75 neurotrophin receptor (p75NTR) in neurons. Therefore, we studied the mechanism of the proNGF/p75NTR pathway in HD-induced neuronal apoptosis. Sprague-Dawley (SD) rats were injected with 400 mg/kg HD once a day for 5 weeks, and VSC4.1 cells were treated with 10, 20, and 40 mM HD in vitro. Results showed that HD effectively induced neuronal apoptosis. Moreover, it up-regulated proNGF and p75NTR levels, activated c-Jun N-terminal kinase (JNK) and c-Jun, and disrupted the balance between B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax). Our findings revealed that the proNGF/p75NTR signaling pathway was involved in HD-induced neuronal apoptosis; it can serve as a theoretical basis for further exploration of the neurotoxic mechanisms of HD.
Asunto(s)
Apoptosis , Hexanonas/farmacología , Factores de Crecimiento Nervioso/metabolismo , Neuronas/metabolismo , Neurotoxinas/farmacología , Precursores de Proteínas/metabolismo , Médula Espinal/efectos de los fármacos , Animales , Línea Celular Tumoral , Hexanonas/toxicidad , MAP Quinasa Quinasa 4/metabolismo , Masculino , Proteínas del Tejido Nervioso/metabolismo , Neuronas/efectos de los fármacos , Neurotoxinas/toxicidad , Ratas , Ratas Sprague-Dawley , Receptores de Factores de Crecimiento/metabolismo , Transducción de Señal , Médula Espinal/citología , Médula Espinal/metabolismoRESUMEN
Differentiation-inducing factor-1 (DIF-1) is a chlorinated alkylphenone (a polyketide) found in the cellular slime mold Dictyostelium discoideum. DIF-1 and its derivative, DIF-1(3M) promote glucose consumption in vitro in mammalian cells and in vivo in diabetic rats; they are expected to be the leading antiobesity and antidiabetes compounds. In this study, we investigated the mechanisms underlying the actions of DIF-1 and DIF-1(3M). In isolated mouse liver mitochondria, these compounds at 2-20 µM promoted oxygen consumption in a dose-dependent manner, suggesting that they act as mitochondrial uncouplers, whereas CP-DIF-1 (another derivative of DIF-1) at 10-20 µM had no effect. In confluent mouse 3T3-L1 fibroblasts, DIF-1 and DIF-1(3M) but not CP-DIF-1 induced phosphorylation (and therefore activation) of AMP kinase (AMPK) and promoted glucose consumption and metabolism. The DIF-induced glucose consumption was reduced by compound C (an AMPK inhibitor) or AMPK knock down. These data suggest that DIF-1 and DIF-1(3M) promote glucose uptake, at least in part, via an AMPK-dependent pathway in 3T3-L1 cells, whereas cellular metabolome analysis revealed that DIF-1 and DIF-1(3M) may act differently at least in part.
Asunto(s)
Adenilato Quinasa/metabolismo , Dictyostelium/metabolismo , Glucosa/metabolismo , Hexanonas/farmacología , Hidrocarburos Clorados/farmacología , Metaboloma/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Pirazoles/farmacología , Pirimidinas/farmacología , Células 3T3 , Adenilato Quinasa/antagonistas & inhibidores , Animales , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Técnicas de Silenciamiento del Gen , Ratones , Mitocondrias/metabolismo , Consumo de Oxígeno/efectos de los fármacos , Fosforilación , ARN Interferente Pequeño , Transducción de Señal/efectos de los fármacosRESUMEN
In maize, nitrate regulates root development thanks to the coordinated action of many players. In this study, the involvement of strigolactones (SLs) and auxin as putative components of the nitrate regulation of lateral root (LR) was investigated. To this aim, the endogenous SL content of maize root in response to nitrate was assessed by liquid chromatography with tandem mass Spectrometry (LC-MS/MS) and measurements of LR density in the presence of analogues or inhibitors of auxin and SLs were performed. Furthermore, an untargeted RNA-sequencing (RNA-seq)-based approach was used to better characterize the participation of auxin and SLs to the transcriptional signature of maize root response to nitrate. Our results suggested that N deprivation induces zealactone and carlactonoic acid biosynthesis in root, to a higher extent if compared to P-deprived roots. Moreover, data on LR density led to hypothesize that the induction of LR development early occurring upon nitrate supply involves the inhibition of SL biosynthesis, but that the downstream target of SL shutdown, besides auxin, also includes additional unknown players. Furthermore, RNA-seq results provided a set of putative markers for the auxin- or SL-dependent action of nitrate, meanwhile also allowing to identify novel components of the molecular regulation of maize root response to nitrate. Globally, the existence of at least four different pathways was hypothesized: one dependent on auxin, a second one mediated by SLs, a third deriving from the SL-auxin interplay, and a last one attributable to nitrate itself through further downstream signals. Further work will be necessary to better assess the reliability of the model proposed.
Asunto(s)
Compuestos Heterocíclicos con 3 Anillos/metabolismo , Ácidos Indolacéticos/metabolismo , Lactonas/metabolismo , Nitratos/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Zea mays/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Germinación , Hexanonas/farmacología , Nitratos/farmacología , Nitrógeno/metabolismo , Orobanchaceae/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Análisis de Secuencia de ARN , Espectrometría de Masas en Tándem , Triazoles/farmacología , Zea mays/efectos de los fármacos , Zea mays/metabolismoRESUMEN
Mesenchymal stem cells (MSCs) are an attractive cell source for tissue regeneration and repair. However, their low differentiation efficacy currently impedes the development of MSC therapy. Therefore, in this study, we investigated the effects of differentiation-inducing factor-1 (DIF-1) on the differentiation efficacy of bone marrow-derived MSCs (BM-MSCs) into adipogenic or osteogenic lineages. BM-MSCs, which were obtained from Sprague-Dawley rats, were positive for the MSC markers (CD29, CD73, and CD90). DIF-1 alone neither affected cell surface antigen expression nor induced adipogenic or osteogenic differentiation. However, DIF-1 significantly enhanced the effects of adipogenic differentiation stimuli, which were evaluated as the number of oil red-O positive cells and the expression of adipocyte differentiation markers (peroxisome proliferator-activated receptor gamma, adipocyte fatty acid-binding protein, and adiponectin). In contrast, DIF-1 significantly attenuated the effects of osteogenic differentiation stimuli, which were evaluated as alizarin red-S positive calcium deposition, and the expression of osteoblast differentiation markers alkaline phosphatase, runt-related transcription factor 2, and osteopontin. We further investigated the mechanism by which DIF-1 affects MSC differentiation efficacy and found that glycogen synthase kinase-3 was the main factor mediating the action of DIF-1 on the adipogenic differentiation of BM-MSCs, whereas it was only partially involved in osteogenic differentiation. These results suggest that DIF-1 supports MSC differentiation toward the desired cell fate by enhancing the differentiation efficacy.
Asunto(s)
Adipogénesis/efectos de los fármacos , Hexanonas/farmacología , Hidrocarburos Clorados/farmacología , Células Madre Mesenquimatosas/metabolismo , Osteogénesis/efectos de los fármacos , Adipocitos/metabolismo , Animales , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Glucógeno Sintasa Quinasa 3/metabolismo , Masculino , Osteoblastos/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacosRESUMEN
The mechanisms by which 2,5-hexanedione (2,5-HD) exposure adversely affects reproduction are unclear. In the present study, whole neonatal mouse ovaries were exposed to 2,5-HD in vitro and then assessed for progesterone levels to determine the effects of this compound on ovary function. Ovarian histomorphological analyses were performed to assess the effects of 2,5-HD on follicular development, and PI3K signaling pathway was evaluated to elucidate the molecular mechanisms of 2,5-HD-mediated toxicity on follicular development. The results showed that after ovarian exposure to 2,5-HD in vitro, the percentage of secondary follicles decreased, while the progesterone levels and the percentage of unhealthy follicles increased, with oocytes identified as the target of damage. The 2,5-HD treatment significantly decreased the of the gene encoding the apoptosis-related protein caspase-8, and PI3K/AKT/FOXO3 pathway signaling was also altered. Furthermore, the effects of 2,5-HD on the gene expression of the PI3K/AKT/FOXO3 and follicular development were blocked by 740Y-P (a PI3K activator), miR-214-3p was abnormally expressed, and luciferase reporter assay results demonstrated that the 3' untranslated region of PI3K was a direct target of miR-214-3p. Overall, the results of the present study indicate that 2,5-HD exposure inhibits follicular development, and the underlying mechanism may involve interference with miR-214-3p-mediated regulation of the PI3K signaling pathway.
Asunto(s)
Hexanonas/farmacología , MicroARNs/metabolismo , Folículo Ovárico/efectos de los fármacos , Ovario/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Animales Recién Nacidos , Apoptosis/efectos de los fármacos , Femenino , Ratones , Oocitos/efectos de los fármacos , Oocitos/metabolismo , Folículo Ovárico/metabolismo , Ovario/metabolismoRESUMEN
Starting from the compound (3,4-dimethoxyphenyl)(2-(3,4-dimethoxyphenyl)cyclopent-1-en-1-yl)methanone (4), two diols and three tetrol derivatives were synthesised. Morover, from the reactions of 1,3-dimethoxybenzene and 1,4-dimethoxybenzene with adipoyl chloride, fifteen new along with nine known compounds were obtained. For the characterizations of compounds, spectroscopic methods such as NMR including DEPT, COSY, HMQC and HMBC experiments and X-ray diffraction were used. The antioxidant activities of novel synthesized seventeen molecules were investigated by analytical methods like ABTSâ¢+ and DPPH⢠scavenging. Also, reducing power these molecules were investigated by Fe3+, Cu2+, and [Fe3+-(TPTZ)2]3+. Some of the molecules record powerful antioxidant profile when compared to putative standards. The inhibition effects of the phenols compounds against AChE and BChE activities were analysed. Also, these phenols were found as effective inhibitors for AChE, hCA I, hCA II, and BChE with Kis in the range of 122.95 ± 18.41-351.31 ± 69.12 nM for hCA I, 62.35 ± 9.03-363.17 ± 180.1 nM for hCA II, 134.57 ± 3.99-457.43 ± 220.10 nM for AChE, and 27.06 ± 9.12-72.98 ± 9.53 nM for BChE, respectively.
Asunto(s)
Antioxidantes/química , Inhibidores de Anhidrasa Carbónica/química , Inhibidores de la Colinesterasa/química , Hexanonas/química , Fenoles/química , Antioxidantes/síntesis química , Antioxidantes/farmacología , Anhidrasa Carbónica I/antagonistas & inhibidores , Anhidrasa Carbónica II/antagonistas & inhibidores , Inhibidores de Anhidrasa Carbónica/síntesis química , Inhibidores de Anhidrasa Carbónica/farmacología , Técnicas de Química Sintética , Inhibidores de la Colinesterasa/síntesis química , Inhibidores de la Colinesterasa/farmacología , Cristalografía por Rayos X , Hexanonas/síntesis química , Hexanonas/farmacología , Humanos , Modelos Moleculares , Fenoles/síntesis química , Fenoles/farmacologíaRESUMEN
Exposure to environmental toxins, including hydrocarbon solvents, increases the risk of developing Parkinson's disease. An emergent hypothesis considers microtubule dysfunction as one of the crucial events in triggering neuronal degeneration in Parkinson's disease. Here, we used 2,5-hexanedione (2,5-HD), the toxic metabolite of n-hexane, to analyse the early effects of toxin-induced neurodegeneration on the cytoskeleton in multiple model systems. In PC12 cells differentiated with nerve growth factor for 5â¯days, we found that 2,5-HD treatment affected all the cytoskeletal components. Moreover, we observed alterations in microtubule distribution and stability, in addition to the imbalance of post-translational modifications of α-tubulin. Similar defects were also found in vivo in 2,5-HD-intoxicated mice. Interestingly, we also found that 2,5-HD exposure induced significant changes in microtubule stability in human skin fibroblasts obtained from Parkinson's disease patients harbouring mutations in PRKN gene, whereas it was ineffective in healthy donor fibroblasts, suggesting that the genetic background may really make the difference in microtubule susceptibility to this environmental Parkinson's disease-related toxin. In conclusion, by showing the imbalance between dynamic and stable microtubules in hydrocarbon-induced parkinsonism, our data support the crucial role of microtubule defects in triggering neurodegeneration.
Asunto(s)
Hexanonas/farmacología , Microtúbulos/efectos de los fármacos , Enfermedad de Parkinson/etiología , Enfermedad de Parkinson/metabolismo , Animales , Línea Celular , Citoesqueleto/efectos de los fármacos , Citoesqueleto/metabolismo , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Ratones , Microtúbulos/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Células PC12 , Trastornos Parkinsonianos/inducido químicamente , Trastornos Parkinsonianos/metabolismo , Ratas , Tubulina (Proteína)/metabolismo , Ubiquitina-Proteína Ligasas/metabolismoRESUMEN
Differentiation-inducing factor-1 (DIF-1), a morphogen produced by the cellular slime mold Dictyostelium discoideum, is a natural product that has attracted considerable attention for its antitumor properties. Here, we report a novel inhibitory effect of DIF-1 on the activation of hepatic stellate cells (HSCs) responsible for liver fibrosis. DIF-1 drastically inhibited transdifferentiation of quiescent HSCs into myofibroblastic activated HSCs in a concentration-dependent manner, thus conferring an antifibrotic effect against in the liver. Neither SQ22536, an adenylate cyclase inhibitor, nor ODQ, a guanylate cyclase inhibitor, showed any effect on the inhibition of HSC activation by DIF-1. In contrast, TWS119, a glycogen synthase kinase 3ß (GSK3ß) inhibitor, attenuated the inhibitory effect of DIF-1. Moreover, the level of inactive GSK3ß (phosphorylated at Ser9) was significantly reduced by DIF-1. DIF-1 also inhibited nuclear translocation of ß-catenin and reduced the level of non-phospho (active) ß-catenin. These results suggest that DIF-1 inhibits HSC activation by disrupting the Wnt/ß-catenin signaling pathway through dephosphorylation of GSK3ß. We propose that DIF-1 is a possible candidate as a therapeutic agent for preventing liver fibrosis.
Asunto(s)
Glucógeno Sintasa Quinasa 3 beta/antagonistas & inhibidores , Células Estrelladas Hepáticas/efectos de los fármacos , Hexanonas/farmacología , Transporte Activo de Núcleo Celular , Adenina/análogos & derivados , Adenina/farmacología , Animales , Antineoplásicos/farmacología , Diferenciación Celular , Transdiferenciación Celular , Dictyostelium , Relación Dosis-Respuesta a Droga , Células Estrelladas Hepáticas/metabolismo , Cirrosis Hepática/metabolismo , Ratones , Oxadiazoles/farmacología , Fosforilación , Pirimidinas/farmacología , Pirroles/farmacología , Quinoxalinas/farmacología , Transducción de Señal , beta Catenina/metabolismoRESUMEN
Differentiation-inducing factor-1 (DIF-1) has been reported to inhibit the proliferation of various mammalian cells by unknown means, although some possible mechanisms of its action have been proposed, including the activation of glycogen synthase kinase-3 (GSK-3). Here, we report an alternative mechanism underlying the action of DIF-1 in human breast cancer cell line MCF-7, on which the effects of DIF-1 have not been examined previously. Intragastric administration of DIF-1 reduced the tumor growth from MCF-7 cells injected into a mammary fat pad of nude mice, without causing adverse effects. In cultured MCF-7, DIF-1 arrested the cell cycle in G0 /G1 phase and suppressed cyclin D1 expression, consistent with our previous results obtained in other cell species. However, DIF-1 did not inhibit the phosphorylation of GSK-3. Investigating an alternative mechanism for the reduction of cyclin D1, we found that DIF-1 reduced the protein levels of signal transducer and activator of transcription 3 (STAT3). The STAT3 inhibitor S3I-201 suppressed cyclin D1 expression and cell proliferation and the overexpression of STAT3 enhanced cyclin D1 expression and accelerated proliferation. Differentiation-inducing factor-1 did not reduce STAT3 mRNA or reduce STAT3 protein in the presence of cycloheximide, suggesting that DIF-1 inhibited STAT3 protein synthesis. Seeking its mechanism, we revealed that DIF-1 inhibited the activation of 70 kDa and/or 85 kDa ribosomal protein S6 kinase (p70S6K /p85S6K ). Inhibition of p70S6K /p85S6K by rapamycin also reduced the expressions of STAT3 and cyclin D1. Therefore, DIF-1 suppresses MCF-7 proliferation by inhibiting p70S6K /p85S6K activity and STAT3 protein synthesis followed by reduction of cyclin D1 expression.
Asunto(s)
Ciclina D1/antagonistas & inhibidores , Hexanonas/farmacología , Hidrocarburos Clorados/farmacología , Proteínas Quinasas S6 Ribosómicas/fisiología , Factor de Transcripción STAT3/antagonistas & inhibidores , Animales , Proliferación Celular/efectos de los fármacos , Ciclina D1/análisis , Femenino , Glucógeno Sintasa Quinasa 3/metabolismo , Humanos , Células MCF-7 , Ratones , Ratones Endogámicos BALB C , Fosforilación , Proteínas Quinasas S6 Ribosómicas 70-kDa , Factor de Transcripción STAT3/biosíntesisRESUMEN
Triple-negative breast cancer (TNBC) is highly proliferative and metastatic, and because it lacks three major molecular targets for chemotherapy (estrogen receptor, progesterone receptor, and human epidermal receptor 2), it is extremely refractory. Differentiation-inducing factor 1 (DIF-1) and DIF-3, which are chlorinated alkylphenones, are lead anticancer compounds found in the cellular slime mold Dictyostelium discoideum. Here, we examined the in vitro effects of DIF-1, DIF-3, and 25 DIF derivatives on cell proliferation and serum-induced cell migration in human MDA-MB-231 cells, a model TNBC cell line. We found that Br-DIF-1, a chlorine-to-bromine-substituted derivative of DIF-1, strongly suppressed cell migration (IC50, 3.8 ïM) with negligible effects on cell proliferation (IC50, >20 ïM). We then synthesized 18 derivatives of Br-DIF-1 and examined the in vitro effects of these derivatives on cell proliferation and serum-induced cell migration in MDA-MB-231 cells. Among the derivatives, Br-DIF-1(+1), Br-DIF-1(+2), and Br-DIF-3(+2) exhibited strong anti-cell migration activities with IC50 values of 1.5, 1.0, and 3.1 ïM, respectively, without affecting cell proliferation (IC50, >20 ïM). These results suggest that these Br-DIF derivatives are good lead compounds for the development of anti-metastatic drugs against TNBC.
Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Dictyostelium/química , Halógenos/farmacología , Hexanonas/farmacología , Hidrocarburos Clorados/farmacología , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Halógenos/química , Hexanonas/síntesis química , Hexanonas/química , Humanos , Hidrocarburos Clorados/síntesis química , Hidrocarburos Clorados/química , Relación Estructura-Actividad , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patologíaRESUMEN
Metabolic profiling and genome mining revealed that anaerobic bacteria have the potential to produce acyloin natural products. In addition to sattazolin A and B, three new sattazolin congeners and a novel acyloin named clostrocyloin were isolated from three strains of Clostridium beijerinckii, a bacterium used for industrial solvent production. Bioactivity profiling showed that the sattazolin derivatives possess antimicrobial activities against mycobacteria and pseudomonads with only low cytotoxicity. Clostrocyloin was found to be mainly active against fungi. The thiamine diphosphate (ThDP)-dependent sattazolin-producing synthase was identified in silico and characterized both in vivo and in in vitro enzyme assays. A related acyloin synthase from the clostrocyloin producer was shown to be responsible for the production of the acyloin core of clostrocyloin. The biotransformation experiments provided first insights into the substrate scope of the clostrocyloin synthase and revealed biosynthetic intermediates.
Asunto(s)
Antibacterianos/química , Antibacterianos/farmacología , Antifúngicos/química , Antifúngicos/farmacología , Alcoholes Grasos/química , Alcoholes Grasos/farmacología , Bacterias Anaerobias/química , Vías Biosintéticas , Clostridium/química , Hexanonas/química , Hexanonas/farmacología , Humanos , Indoles/química , Indoles/farmacología , Mycobacterium/efectos de los fármacos , Infecciones por Mycobacterium/tratamiento farmacológico , Micosis/tratamiento farmacológico , Pseudomonas/efectos de los fármacos , Infecciones por Pseudomonas/tratamiento farmacológicoRESUMEN
Longhorn beetles are ecologically important insects in forest ecosystems as decomposers of woody substrates, microhabitat engineers, and as components of forest food webs. These species can be greatly affected both positively and negatively by modern forestry management practices, and should be monitored accordingly. Through headspace sampling, coupled gas chromatography-electroantennography, gas chromatography-mass spectrometry, and field bioassays, we identified two compounds, 2-methyl-1-butanol and 3-hydroxy-2-hexanone, that constitute aggregation-sex pheromone attractants of three cerambycid species which breed primarily in different types of fresh, recently dead oak wood in Northern Europe: Pyrrhidium sanguineum (L.), Phymatodes alni ssp. alni (L.), and Phymatodes testaceus (L.) (Cerambycinae: Callidiini). Analyses of headspace volatiles collected from live insects indicated that the male-produced aggregation-sex pheromone of P. sanguineum is a 1-15:100 blend of (R)-2-methyl-1-butanol and (R)-3-hydroxy-2-hexanone, whereas the corresponding ratios for P. alni were 70-110:100. In field bioassays, adult P. sanguineum and P. alni were significantly attracted to multiple blends with varying ratios of the two compounds. When tested individually, the compounds were minimally attractive. In contrast, adult P. testaceus exhibited nonspecific attraction to both of the individual compounds and to different blends, despite the hydroxyketone not being part of its pheromone, which consists of (R)-2-methyl-1-butanol alone. Overall, our results suggest that a blend of 50:100 of racemic 2-methyl-1-butanol and 3-hydroxy-2-hexanone is appropriate for parallel, cost-efficient pheromone-based monitoring of all three species. In particular, these species could serve as useful indicators of how modern forestry practices affect a whole guild of saproxylic insects that require ephemeral deadwood substrates for successful breeding.
Asunto(s)
Escarabajos/fisiología , Atractivos Sexuales/química , Animales , Europa (Continente) , Femenino , Cromatografía de Gases y Espectrometría de Masas , Hexanonas/análisis , Hexanonas/farmacología , Masculino , Pentanoles/análisis , Pentanoles/farmacología , Atractivos Sexuales/farmacología , Conducta Sexual Animal/efectos de los fármacos , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/farmacologíaRESUMEN
At the end of its life cycle, the cellular slime mold Dictyostelium discoideum forms a fruiting body consisting of spores and a multicellular stalk. Originally, the chlorinated alkylphenone differentiation-inducing factors (DIFs) -1 and -3 were isolated as stalk cell inducers in D. discoideum. Later, DIFs and their derivatives were shown to possess several biologic activities including antitumor and anti-Trypanosoma properties. In this study, we examined the antibacterial activities of approximately 30 DIF derivatives by using several bacterial species. Several of the DIF derivatives strongly suppressed the growth of the Gram-positive bacteria Staphylococcus aureus, Bacillus subtilis, and Enterococcus faecalis and Enterococcus faecium, at minimum inhibitory concentrations (MICs) in the sub-micromolar to low-micromolar range. In contrast, none of the DIF derivatives evaluated had any noteworthy effect on the growth of the Gram-negative bacterium Escherichia coli (MIC, >100 µM). Most importantly, several of the DIF derivatives strongly inhibited the growth of methicillin-resistant S. aureus and vancomycin-resistant E. faecalis and E. faecium. Transmission electron microscopy revealed that treatment with DIF derivatives led to the formation of distinct multilayered structures consisting of cell wall or plasma membrane in S. aureus. The present results suggest that DIF derivatives are good lead compounds for developing novel antimicrobials.
Asunto(s)
Antibacterianos/farmacología , Diferenciación Celular/efectos de los fármacos , Dictyostelium/citología , Hexanonas/farmacología , Antibacterianos/química , Bacterias/efectos de los fármacos , Bacterias/ultraestructura , Dibenzofuranos/química , Dibenzofuranos/farmacología , Dictyostelium/efectos de los fármacos , Hexanonas/química , Pruebas de Sensibilidad MicrobianaRESUMEN
Strigolactones (SLs) are carotenoid-derived plant hormones that also act in the rhizosphere to stimulate germination of root-parasitic plants and enhance plant symbiosis with beneficial microbes. Here, the role of SLs was investigated in the interaction of rice (Oryza sativa) roots with the root-knot nematode Meloidogyne graminicola. Genetic approaches and chemical sprays were used to manipulate SL signaling in rice before infection with M. graminicola. Then, nematode performance was evaluated and plant defense hormones were quantified. Meloidogyne graminicola infection induced SL biosynthesis and signaling and suppressed jasmonic acid (JA)-based defense in rice roots, suggesting a potential role of SLs during nematode infection. Whereas the application of a low dose of the SL analogue GR24 increased nematode infection and decreased jasmonate accumulation, the SL biosynthesis and signaling d mutants were less susceptible to M. graminicola, and constitutively accumulated JA and JA-isoleucine compared with wild-type plants. Spraying with 0.1 µM GR24 restored nematode susceptibility in SL-biosynthesis mutants but not in the signaling mutant. Furthermore, foliar application of the SL biosynthesis inhibitor TIS108 impeded nematode infection and increased jasmonate levels in rice roots. In conclusion, SL signaling in rice suppresses jasmonate accumulation and promotes root-knot nematode infection.