RESUMEN
Freesia refracta (FR), a perennial flower of the Iris family (Iridaceae), is widely used in cosmetics despite limited scientific evidence of its skin benefits and chemical composition, particularly of FR callus extract (FCE). This study identified biologically active compounds in FCE and assessed their skin benefits, focusing on anti-aging. FR calli were cultured, extracted with water at 40 °C, and analyzed using Centrifugal Partition Chromatography (CPC), Nuclear Magnetic Resonance (NMR), and HCA, revealing key compounds, namely nicotinamide and pyroglutamic acid. FCE significantly increased collagen I production by 52% in normal and aged fibroblasts and enhanced fibroblast-collagen interaction by 37%. An in vivo study of 43 female volunteers demonstrated an 11.1% reduction in skin roughness and a 2.3-fold increase in collagen density after 28 days of cream application containing 3% FCE. Additionally, the preservation tests of cosmetics containing FCE confirmed their stability over 12 weeks. These results suggest that FCE offers substantial anti-aging benefits by enhancing collagen production and fibroblast-collagen interactions. These findings highlighted the potential of FCE in cosmetic applications, providing significant improvements in skin smoothness and overall appearance. This study fills a gap in the scientific literature regarding the skin benefits and chemical composition of FR callus extract, supporting its use in the development of effective cosmeceuticals.
Asunto(s)
Fibroblastos , Estrés Oxidativo , Extractos Vegetales , Envejecimiento de la Piel , Piel , Envejecimiento de la Piel/efectos de los fármacos , Extractos Vegetales/farmacología , Extractos Vegetales/química , Humanos , Femenino , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Piel/metabolismo , Piel/efectos de los fármacos , Transcriptoma/efectos de los fármacos , Adulto , Colágeno/metabolismo , Cosméticos/farmacología , Persona de Mediana Edad , Niacinamida/farmacología , Ácido Pirrolidona Carboxílico/análogos & derivados , Ácido Pirrolidona Carboxílico/farmacología , Ácido Pirrolidona Carboxílico/metabolismoRESUMEN
Recombinant adeno-associated virus (rAAV)-based gene therapies offer an immense opportunity for rare diseases, such as amyotrophic lateral sclerosis (ALS), which is defined by the loss of the upper and the lower motor neurons. Here, we describe generation, characterization, and utilization of a novel vector system, which enables expression of the active form of hepatocyte growth factor (HGF) under EF-1α promoter with bovine growth hormone (bGH) poly(A) sequence and is effective with intrathecal injections. HGF's role in promoting motor neuron survival had been vastly reported. Therefore, we investigated whether intrathecal delivery of HGF would have an impact on one of the most common pathologies of ALS: the TDP-43 pathology. Increased astrogliosis, microgliosis and progressive upper motor neuron loss are important consequences of ALS in the motor cortex with TDP-43 pathology. We find that cortex can be modulated via intrathecal injection, and that expression of HGF reduces astrogliosis, microgliosis in the motor cortex, and help restore ongoing UMN degeneration. Our findings not only introduce a novel viral vector for the treatment of ALS, but also demonstrate modulation of motor cortex by intrathecal viral delivery, and that HGF treatment is effective in reducing astrogliosis and microgliosis in the motor cortex of ALS with TDP-43 pathology.
Asunto(s)
Esclerosis Amiotrófica Lateral , Corteza Motora , Animales , Bovinos , Humanos , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/terapia , Proteínas de Unión al ADN/genética , Gliosis , Factor de Crecimiento de Hepatocito/genética , Corteza Motora/patologíaRESUMEN
Hepatocyte growth factor (HGF) is a multifunctional protein that plays a critical role in the angiogenic, neurotrophic, antifibrotic, and antiapoptotic activities of various cell types. It has been previously reported that intramuscular injection of pCK-HGF-X7 (or VM202), a plasmid DNA designed to express both native isoforms of human HGF (Pyun et al., 2010), significantly reduced the level of neuropathic pain in clinical studies as well as in a variety of animal models. In clinical studies, it has been observed that pCK-HGF-X7 appeared to give much higher pain-relieving effects in subjects not taking pregabalin or gabapentin, α2δ1 calcium channel blockers frequently prescribed for reducing pain in patients with diabetic peripheral neuropathy. In this study, we tested the effects of gabapentin on HGF-mediated pain reduction and nerve regeneration in vivo. Consistent with the data from clinical studies, gabapentin administration inhibited the pain reduction and axon regeneration effects mediated by HGF expression from pCK-HGF-X7. In the context of nerve regenerative effects, treatment with gabapentin or EGTA, a Ca2+ chelator, inhibited HGF-mediated axon outgrowth of injured sciatic nerves in vivo. Taken together, i.m. injection of HGF-encoding plasmid DNA ameliorated pain symptoms and enhanced the regeneration of injured nerves, and these therapeutic effects of HGF were significantly hindered by gabapentin treatment, suggesting the possible involvement of Ca2+ in the pro-regenerative activities of native HGF derived from treatment with pCK-HGF-X7.
Asunto(s)
Neuralgia , Traumatismos de los Nervios Periféricos , Analgésicos/farmacología , Analgésicos/uso terapéutico , Animales , Axones/metabolismo , ADN/genética , ADN/farmacología , Modelos Animales de Enfermedad , Gabapentina/farmacología , Gabapentina/uso terapéutico , Terapia Genética , Factor de Crecimiento de Hepatocito/genética , Factor de Crecimiento de Hepatocito/farmacología , Factor de Crecimiento de Hepatocito/uso terapéutico , Humanos , Regeneración Nerviosa , Neuralgia/tratamiento farmacológico , Traumatismos de los Nervios Periféricos/tratamiento farmacológicoRESUMEN
Hepatocyte growth factor (HGF) is a secretory protein that is involved in various biological activities such as angiogenesis, neuroprotection, and anti-inflammatory effects. Intramuscular injection of an HGF-encoding plasmid DNA (pCK-HGF-X7) has been shown to produce pain-relieving effects in a rodent model and patients with neuropathic pain.To further investigate the underlying mechanism, we investigated the anti-inflammatory effects of HGF in the context of neuropathic pain. Consistent with previous data, intramuscular injection of pCK-HGF-X7 showed pain relieving effects up to 8 weeks and pharmacological blockade of the c-Met receptor hindered this effect, which suggest that the analgesic effect was c-Met receptor-dependent. At the histological level, macrophage infiltration in the dorsal root ganglion (DRG) was significantly decreased in the pCK-HGF-X7 injected group. Moreover, HGF treatment significantly downregulated the LPS-mediated induction of pro-inflammatory cytokines in primary cultured DRG neurons. Taken together, these data suggest that HGF-encoding plasmid DNA attenuates neuropathic pain via controlling the expression of pro-inflammatory cytokines.
Asunto(s)
Factor de Crecimiento de Hepatocito , Neuralgia , Animales , Antiinflamatorios/metabolismo , ADN/metabolismo , Modelos Animales de Enfermedad , Ganglios Espinales/metabolismo , Terapia Genética , Factor de Crecimiento de Hepatocito/metabolismo , Humanos , Inyecciones Intramusculares , Neuralgia/genética , Neuralgia/metabolismo , Neuralgia/terapia , Plásmidos/genéticaRESUMEN
We aimed to investigate the role of cMet agonistic antibody (cMet Ab) in preventing kidney fibrosis during acute kidney injury (AKI) to chronic kidney disease (CKD) transition. Additionally, we explored the effect of cMet Ab on TGF-ß1/Smad pathway during the pathogenesis of kidney fibrosis. A unilateral ischemia-reperfusion injury (UIRI) mouse model was established to induce AKI-to-CKD transition. Furthermore, we incubated human proximal tubular epithelial cells (hPTECs) under hypoxic conditions as in vitro model of kidney fibrosis. We analyzed the soluble plasma cMet level in patients with AKI requiring dialysis. Patients who did not recover kidney function and progressed to CKD presented a higher increase in the cMet level. The kidneys of mice treated with cMet Ab showed fewer contractions and weighed more than the controls. The mice in the cMet Ab-treated group showed reduced fibrosis and significantly decreased expression of fibronectin and α-smooth muscle actin. cMet Ab treatment decreased inflammatory markers (MCP-1, TNF-α, and IL-1ß) expression, reduced Smurf1 and Smad2/3 level, and increased Smad7 expressions. cMet Ab treatment increased cMet expression and reduced the hypoxia-induced increase in collagen-1 and ICAM-1 expression, thereby reducing apoptosis in the in vitro cell model. After cMet Ab treatment, hypoxia-induced expression of Smurf1, Smad2/3, and TGF-ß1 was reduced, and suppressed Smad7 was activated. Down-regulation of Smurf1 resulted in suppression of hypoxia-induced fibronectin expression, whereas treatment with cMet Ab showed synergistic effects. cMet Ab can successfully prevent fibrosis response in UIRI models of kidney fibrosis by decreasing inflammatory response and inhibiting the TGF-ß1/Smad pathway.
Asunto(s)
Lesión Renal Aguda/patología , Insuficiencia Renal Crónica/metabolismo , Proteína smad7/metabolismo , Lesión Renal Aguda/tratamiento farmacológico , Lesión Renal Aguda/metabolismo , Animales , Fibrosis/patología , Humanos , Riñón/metabolismo , Ratones Endogámicos C57BL , Insuficiencia Renal Crónica/tratamiento farmacológico , Insuficiencia Renal Crónica/patología , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Acute kidney injury (AKI) is a very common complication with high morbidity and mortality rates and no fundamental treatment. In this study, we investigated whether the hepatocyte growth factor (HGF)/cMet pathway is associated with the development of AKI and how the administration of a cMet agonistic antibody (Ab) affects an AKI model. In the analysis using human blood samples, cMet and HGF levels were found to be significantly increased in the AKI group, regardless of underlying renal function. The administration of a cMet agonistic Ab improved the functional and histological changes after bilateral ischaemia-reperfusion injury. TUNEL-positive cells and Bax/Bcl-2 ratio were also reduced by cMet agonistic Ab treatment. In addition, cMet agonistic Ab treatment significantly increased the levels of PI3K, Akt and mTOR. Furthermore, after 24 hours of hypoxia induction in human proximal tubular epithelial cells, treatment with the cMet agonistic Ab also showed dose-dependent antiapoptotic effects similar to those of the recombinant HGF treatment. Even when the HGF axis was blocked with a HGF-blocking Ab, the cMet agonistic Ab showed an independent dose-dependent antiapoptotic effect. In conclusion, cMet expression is associated with the occurrence of AKI. cMet agonistic Ab treatment attenuates the severity of AKI through the PI3K/Akt/mTOR pathway and improves apoptosis. cMet agonistic Ab may have important significance for the treatment of AKI.
Asunto(s)
Lesión Renal Aguda/metabolismo , Anticuerpos Monoclonales/farmacología , Apoptosis/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-met/antagonistas & inhibidores , Daño por Reperfusión/metabolismo , Lesión Renal Aguda/diagnóstico , Lesión Renal Aguda/etiología , Anciano , Animales , Ciclo Celular/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Daño por Reperfusión/diagnóstico , Daño por Reperfusión/etiología , Índice de Severidad de la Enfermedad , Transducción de Señal/efectos de los fármacos , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
The prediction of prognosis in patients with immunoglobulin A nephropathy (IgAN) is challenging. We investigated the correlation between urinary cMet (ucMet) levels and clinical parameters and examined the effects of cMet agonistic antibody (cMet Ab) in an in vitro IgAN model. Patients diagnosed with IgAN (n = 194) were divided into three groups representing undetectable (Group 1), below-median (Group 2) and above-median (Group 3) levels of ucMet/creatinine (ucMet/Cr). Stained kidney biopsy samples were graded according to cMet intensity. Primary-cultured human mesangial cells were stimulated with recombinant tumour necrosis factor (TNF)-α and treated with cMet Ab. Our results showed that ucMet/Cr levels positively correlated with proteinuria (P < .001). During the follow-up, patients in Group 3 showed a significantly lower probability of complete remission (CR; uPCr < 300 mg/g) than those in groups 1 and 2, after adjusting for blood pressure, estimated glomerular filtration rate, and proteinuria, which influence clinical prognosis (HR 0.60, P = .038); moreover, ucMet/Cr levels were also associated with glomerular cMet expression. After TNF-α treatment, the proliferation of mesangial cells and increased interleukin-8 and intercellular adhesion molecule-1 expression were markedly reduced by cMet Ab in vitro. In conclusion, ucMet/Cr levels significantly correlated with proteinuria, glomerular cMet expression, and the probability of CR. Further, cMet Ab treatment alleviated the inflammation and proliferation of mesangial cells. Hence, ucMet could serve as a clinically significant marker for treating IgAN.
Asunto(s)
Glomerulonefritis por IGA/orina , Proteínas Proto-Oncogénicas c-met/orina , Adulto , Biomarcadores/orina , Creatinina/orina , Femenino , Glomerulonefritis por IGA/complicaciones , Humanos , Riñón/patología , Masculino , Persona de Mediana Edad , Modelos Biológicos , Pronóstico , Proteinuria/complicaciones , Inducción de RemisiónRESUMEN
Hepatocyte growth factor (HGF) is a versatile neurotrophic factor that mediates a variety of cellular activities. In this study, we investigated the effects of intramuscularly injected recombinant AAV vectors expressing HGF in two pathologic conditions: the sciatic nerve crush and the SOD1-G93A transgenic mouse models. AAV serotype 6 (rAAV6) was chosen based on its expression levels in, and capability of moving to, the spinal cord from the injected muscle area. In the nerve crush model, rAAV6-HGF was shown to reduce the degree of mechanical allodynia, increase the cross-sectional area of muscle fibers, promote regrowth of peripheral axons, and improve motor functions. In the SOD1-G93A TG mouse model, rAAV6-HGF increased the mass of the tibialis anterior and gastrocnemius, alleviated disease symptoms, and prolonged survival. Improvements in integrity and functions of muscle in these models seemed to have come from the ability of HGF produced from rAAV6-HGF to regulate the expression of various atrogenes through the control of the FOXO signaling pathway. Our findings suggested that intramuscular injection of rAAV6-HGF might be used to relieve various symptoms associated with muscle atrophy and/or nerve damages observed in a majority of neuromuscular diseases.
Asunto(s)
Dependovirus/genética , Técnicas de Transferencia de Gen , Factor de Crecimiento de Hepatocito/genética , Músculo Esquelético/metabolismo , Unión Neuromuscular/metabolismo , Superóxido Dismutasa-1/genética , Animales , Dependovirus/metabolismo , Proteína Forkhead Box O1/genética , Proteína Forkhead Box O1/metabolismo , Expresión Génica , Vectores Genéticos/administración & dosificación , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Fuerza de la Mano/fisiología , Factor de Crecimiento de Hepatocito/metabolismo , Hiperalgesia/genética , Hiperalgesia/metabolismo , Hiperalgesia/fisiopatología , Hiperalgesia/prevención & control , Inyecciones Intramusculares , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Neuronas Motoras/metabolismo , Neuronas Motoras/patología , Músculo Esquelético/inervación , Músculo Esquelético/patología , Mutación , Compresión Nerviosa/métodos , Unión Neuromuscular/patología , Prueba de Desempeño de Rotación con Aceleración Constante , Nervio Ciático/lesiones , Nervio Ciático/metabolismo , Nervio Ciático/patología , Superóxido Dismutasa-1/deficienciaRESUMEN
Hepatocyte growth factor (HGF) is a multifunctional protein that contains angiogenic and neurotrophic properties. In the current study, we investigated the analgesic effects of HGF by using a plasmid DNA that was designed to express 2 isoforms of human HGF-pCK-HGF-X7 (or VM202)-in a chronic constriction injury (CCI) -induced mouse neuropathic pain model. Intramuscular injection of pCK-HGF-X7 into proximal thigh muscle induced the expression of HGF in the muscle, sciatic nerve, and dorsal root ganglia (DRG). This gene transfer procedure significantly attenuated mechanical allodynia and thermal hyperalgesia after CCI. Injury-induced expression of activating transcription factor 3, calcium channel subunit α2δ1, and CSF1 in the ipsilateral DRG neurons was markedly down-regulated in the pCK-HGF-X7-treated group, which suggested that HGF might exert its analgesic effects by inhibiting pain-mediating genes in the sensory neurons. In addition, suppressed CSF1 expression in DRG neurons by pCK-HGF-X7 treatment was accompanied by a noticeable suppression of the nerve injury-induced glial cell activation in the spinal cord dorsal horn. Taken together, our data show that pCK-HGF-X7 attenuates nerve injury-induced neuropathic pain by inhibiting pain-related factors in DRG neurons and subsequent spinal cord glial activation, which suggests its therapeutic efficacy in the treatment of neuropathic pain.-Nho, B., Lee, J., Lee, J., Ko, K. R., Lee, S. J., Kim, S. Effective control of neuropathic pain by transient expression of hepatocyte growth factor in a mouse chronic constriction injury model.
Asunto(s)
Factor de Crecimiento de Hepatocito/genética , Neuralgia/genética , Neuralgia/terapia , Traumatismos de la Médula Espinal/genética , Animales , Canales de Calcio/genética , Constricción , Modelos Animales de Enfermedad , Regulación hacia Abajo/genética , Ganglios Espinales/metabolismo , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Humanos , Hiperalgesia/genética , Factor Estimulante de Colonias de Macrófagos/genética , Masculino , Ratones , Ratones Endogámicos ICR , Músculo Esquelético/metabolismo , Neuroglía/metabolismo , Nervio Ciático/metabolismo , Células Receptoras Sensoriales/metabolismo , Asta Dorsal de la Médula Espinal/metabolismo , Factor de Transcripción 3/genéticaRESUMEN
Hepatocyte growth factor (HGF) receptor is a member of the receptor tyrosine kinases (RTKs) and has been reported to perform diverse functions in various cell types during both the developmental and adult stages. Among different roles, HGF is best known for its angiogenic effects of inducing the migration of endothelial cells. Because angiogenesis is one of the prerequisite steps for tumor metastasis, HGF-dependent cell migration has to be tightly controlled. However, the underlying mechanisms regulating the optimum level of HGF/c-met signaling have been poorly understood. In this study, we tested whether the migration of endothelial cells is regulated by a negative feedback mechanism under disproportionately large amounts of HGF. Data from endothelial cell migration assays showed that HGF activity increased as its concentration increased, but declined beyond a certain point. Under limiting conditions, amounts of phosphorylated Erk and Akt surged, reaching a plateau in which the enhanced level was more or less maintained. The c-met receptor was degraded when unnecessarily large amounts of HGF were present. Under these conditions, HGF could no longer activate downstream signaling pathways even if cells were re-treated with optimal amounts of HGF. Excessive doses of HGF increased the phosphorylation of tyrosine residue 1003 involved in the ubiquitination of c-met, and phosphorylated c-met was diverted toward the proteasomal degradation pathway. Taken together, HGF/c-met signaling is tightly regulated by a negative feedback loop through an ubiquitin-proteasomal degradation pathway.
Asunto(s)
Factor de Crecimiento de Hepatocito/farmacología , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteínas Proto-Oncogénicas c-met/metabolismo , Movimiento Celular , Células Endoteliales , Retroalimentación Fisiológica , Humanos , Fosforilación , UbiquitinaciónRESUMEN
We previously reported that the expression of hepatocyte growth factor (HGF) was highly induced after peripheral nerve damage, and that c-Fos is one of many cellular genes whose expressions are affected by the increased level of HGF[1]. c-Fos is an important component of AP-1 heterodimer, but its role has not been clearly understood in the context of HGF and Schwann cells (SCs). In this study, we investigated the relationship between HGF and c-Fos. First, it was confirmed that the c-Fos was increased in SCs after nerve injury, while this effect abrogated by PHA-665752, an inhibitor of c-met receptor. When primary SCs were treated with recombinant HGF protein, c-Fos expression was regulated in a typical quick, transient fashion at both RNA and proteins levels. HGF-mediated induction of c-Fos expression was highly suppressed by specific inhibitors of ERK and CREB, respectively. The knock down of c-Fos expression by siRNA almost completely blocked various HGF-mediated effects in SCs, such as induction of gene expression of GDNF, LIF, and c-Myc, and migration of SCs, indicating that c-Fos might play a key role in HGF effects. Taken together, our results suggested that c-Fos plays a key role(s) in HGF-mediated effects on neurotrophic genes and cell migration.
Asunto(s)
Movimiento Celular/genética , Regulación de la Expresión Génica , Factor de Crecimiento de Hepatocito/genética , Proteínas Proto-Oncogénicas c-fos/genética , Células de Schwann/metabolismo , Nervio Ciático/metabolismo , Animales , Butadienos/farmacología , Movimiento Celular/efectos de los fármacos , Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Factor de Crecimiento de Hepatocito/metabolismo , Factor de Crecimiento de Hepatocito/farmacología , Indoles/farmacología , Factor Inhibidor de Leucemia/genética , Factor Inhibidor de Leucemia/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Compresión Nerviosa/métodos , Nitrilos/farmacología , Cultivo Primario de Células , Proteínas Proto-Oncogénicas c-fos/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-fos/metabolismo , Proteínas Proto-Oncogénicas c-met/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-met/genética , Proteínas Proto-Oncogénicas c-met/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Células de Schwann/citología , Células de Schwann/efectos de los fármacos , Nervio Ciático/lesiones , Transducción de Señal , Sulfonas/farmacología , Factor de Transcripción AP-1/genética , Factor de Transcripción AP-1/metabolismoRESUMEN
A series of 4-aryl-thieno[1,4]diazepin-2-one were synthesized and evaluated for their antiproliferative activities against the A375P melanoma and U937 hematopoietic cell lines. Several compounds showed very potent antiproliferative activities toward both cell lines and the activities were better than that of sorafenib, the reference standard. Derivatives were made as amide (8a-8i, 9a-9m) and urea (10a-10d, 11a-11d) with diverse hydrophobic moieties. One of the most potent inhibitor 10d, 1-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-3-(4-(2-oxo-2,3-dihydro-1H-thieno [3,4-b][1,4]diazepin-4-yl)phenyl)urea was found to be very potent inhibitor of multi-protein kinases including FMS kinase (IC50â¯=â¯3.73â¯nM) and is a promising candidate for further development in therapeutics for cancer.
Asunto(s)
Antineoplásicos/farmacología , Azepinas/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Quinasas/metabolismo , Tiofenos/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Azepinas/síntesis química , Azepinas/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Descubrimiento de Drogas , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Relación Estructura-Actividad , Tiofenos/síntesis química , Tiofenos/química , Células U937RESUMEN
The recent success of vemurafenib shows the importance of selective BRAF V600E inhibition in melanoma. However, paradoxical activation by structurally diverse ATP-competitive RAF kinase inhibitors strongly suggests that selective CRAF inhibitors, not BRAF inhibitors, would be ideal for some Ras mutation cancer treatment. In this respect, we approached designing selective CRAF inhibitors starting from in silico fragment screening and synthesized a 3-carboxamido-2H-indazole-6-arylamide scaffold. Most of the compounds showed potent antiproliferative activity against the WM3629 melanoma cell line and the most promising compound, compound 10d, was found to be a potent and selective CRAF inhibitor with an IC50 value of 38.6 nM, which shows greater than 270-fold selectivity over BRAF kinase (9.45 µM).
Asunto(s)
Amidas/química , Diseño de Fármacos , Indazoles/química , Inhibidores de Proteínas Quinasas/síntesis química , Proteínas Proto-Oncogénicas c-raf/antagonistas & inhibidores , Amidas/síntesis química , Amidas/farmacología , Sitios de Unión , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , Simulación del Acoplamiento Molecular , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/toxicidad , Estructura Terciaria de Proteína , Proteínas Proto-Oncogénicas c-raf/metabolismoRESUMEN
The root serves as an essential organ in plant growth by taking up nutrients and water from the soil and supporting the rest of the plant body. Some plant species utilize roots as storage organs. Sweet potatoes (Ipomoea batatas), cassava (Manihot esculenta), and radish (Raphanus sativus), for example, are important root crops. However, how their root growth is regulated remains unknown. In this study, we characterized the relationship between cambium and radial root growth in radish. Through a comparative analysis with Arabidopsis root expression data, we identified putative cambium-enriched transcription factors in radish and analysed their expression in representative inbred lines featuring distinctive radial growth. We found that cell proliferation activities in the cambium positively correlated with radial growth and final yields of radish roots. Expression analysis of candidate transcription factor genes revealed that some genes are differentially expressed between inbred lines and that the difference is due to the distinct cytokinin response. Taken together, we have demonstrated for the first time, to the best of our knowledge, that cytokinin-dependent radial growth plays a key role in the yields of root crops.
Asunto(s)
Citocininas/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Raphanus/crecimiento & desarrollo , Raphanus/genética , Factores de Transcripción/genética , Biomasa , Cámbium/citología , Cámbium/genética , Cámbium/crecimiento & desarrollo , División Celular , Regulación del Desarrollo de la Expresión Génica , Proteínas de Plantas/metabolismo , Raíces de Plantas/citología , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raphanus/metabolismo , Factores de Transcripción/metabolismoRESUMEN
Oxidative stress is induced by the accumulation of free radicals, resulting in an imbalanced cellular redox state, which has been implicated in a variety of human diseases. Dehydrodiconiferyl alcohol (DHCA), a lignan compound isolated from Cucurbita moschata, has previously been reported to contain anti-adipogenic and anti-lipogenic effects on 3T3-L1 cells and primary MEFs (Abraham and Kappas, 2008). In this study, it was tested whether DHCA could affect the expression of HO-1, using Raw264.7 mouse macrophage cell line. DHCA increased the protein and RNA levels of HO-1 and upregulated its promoter activity. Data from transient transfection assays indicated that ARE located in the E1 region of the HO-1 promoter are important in this DHCA-mediated induction of HO-1 expression. DHCA was also shown to enhance the nuclear translocation and binding of Nrf2 to the respective DNA sequences. The upregulation of HO-1 expression by DHCA was also observed in primary macrophages derived from wild type animals, but not in those from Nrf2 KO mice. Effects of DHCA on HO-1 and Nrf2 were reduced when cells were treated with an AMPK inhibitor, Compound C, but not by PI3K/Akt or MAPK inhibitors. Data from an experiment using a specific siRNA or chemical inhibitor for HO-1 suggested that the DHCA-mediated induction of the HO-1 protein could suppress the LPS-stimulated production of NO. Taken together, our data suggest that DHCA induces the expression of HO-1 by controlling its promoter activity through the AMPK-Nrf2 pathway, eventually leading to the reduction of NO production, and may thus have potential as an effective antioxidant.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Regulación Enzimológica de la Expresión Génica , Hemo-Oxigenasa 1/biosíntesis , Proteínas de la Membrana/biosíntesis , Factor 2 Relacionado con NF-E2/metabolismo , Fenoles/toxicidad , Regulación hacia Arriba/efectos de los fármacos , Células 3T3-L1 , Proteínas Quinasas Activadas por AMP/antagonistas & inhibidores , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Hemo-Oxigenasa 1/genética , Masculino , Proteínas de la Membrana/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Factor 2 Relacionado con NF-E2/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Regulación hacia Arriba/fisiologíaRESUMEN
The synthesis of a novel series of (4-aminobenzyl/benzoyl)-1H-imidazol-1-yl pyrimidin-2-yl derivatives 9, 10, 18, 19 and their in vitro antiproliferative activities against the A375P human melanoma cell line and the U937 human leukemic monocyte lymphoma cell line are described. Potent antiproliferative effects were found from 9l, 9s and 10c; 10c was found to be a highly potent and selective BRAF V600E and CRAF inhibitor (IC50=38.3 nM and 8.79 nM).
Asunto(s)
Antineoplásicos/farmacología , Diseño de Fármacos , Compuestos de Fenilurea/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas B-raf/antagonistas & inhibidores , Pirimidinas/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Modelos Moleculares , Estructura Molecular , Compuestos de Fenilurea/síntesis química , Compuestos de Fenilurea/química , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Proteínas Proto-Oncogénicas B-raf/metabolismo , Pirimidinas/síntesis química , Pirimidinas/química , Relación Estructura-Actividad , Células U937RESUMEN
We report a case of a 45-year-old man presenting with tachycardia and palpitation. Echocardiography indicated severe tricuspid regurgitation. We suspected traumatic tricuspid damage due to high energy trauma in a motor vehicle accident 17â¯years earlier. He underwent a sternotomy, and his tricuspid valve was repaired with chordal reconstruction, indentation closure, and ring annuloplasty. The postoperative period was uneventful, and he was discharged 10â¯days after the operation. This report highlights the value of echocardiography for diagnosis of primary tricuspid regurgitation related to trauma, and the importance of early diagnosis to allow surgical intervention before irreversible damage occurs. Learning objective: Traumatic tricuspid regurgitation is a rare cardiovascular complication of blunt chest trauma. The mechanism of the tricuspid valve injury is thought to be secondary to sudden impact causing an anteroposterior compression of the right ventricle by the sternum in end-diastole. This injury is often incidentally identified or can be missed until the patient experiences symptoms of right heart failure resulting from severe tricuspid regurgitation.
RESUMEN
A water-soluble extract from the stems of Cucurbita moschata, code named PG105, was previously found to contain strong anti-obesity activities in a high fat diet-induced obesity mouse model. One of its biological characteristics is that it inhibits 3T3-L1 adipocyte differentiation. To isolate the biologically active compound(s), conventional solvent fractionation was performed, and the various fractions were tested for anti-adipogenic activity using Oil Red O staining method. A single spot on thin layer chromatography of the chloroform fraction showed a potent anti-adipogenic activity. When purified, the structure of its major component was resolved as dehydrodiconiferyl alcohol (DHCA), a lignan, by NMR and mass spectrometry analysis. In 3T3-L1 cells, synthesized DHCA significantly reduced the expression of several adipocyte marker genes, including peroxisome proliferator-activated receptor γ (Pparg), CCAAT/enhancer-binding protein α (Cebpa), fatty acid-binding protein 4 (Fabp4), sterol response element-binding protein-1c (Srebp1c), and stearoyl-coenzyme A desaturase-1 (Scd), and decreased lipid accumulation without affecting cell viability. DHCA also suppressed the mitotic clonal expansion of preadipocytes (an early event of adipogenesis), probably by suppressing the DNA binding activity of C/EBPß, and lowered the production level of cyclinA and cyclin-dependent kinase 2 (Cdk2), coinciding with the decrease in DNA synthesis and cell division. In addition, DHCA directly inhibited the expression of SREBP-1c and SCD-1. Similar observations were made, using primary mouse embryonic fibroblasts. Taken together, our data indicate that DHCA may contain dual activities, affecting both adipogenesis and lipogenesis.
Asunto(s)
Adipogénesis/efectos de los fármacos , Cucurbita/química , Fibroblastos/citología , Lipogénesis/efectos de los fármacos , Obesidad/metabolismo , Fenoles/farmacología , Extractos Vegetales/farmacología , Células 3T3-L1 , Adipocitos/citología , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Animales , Proteína alfa Potenciadora de Unión a CCAAT/genética , Proteína alfa Potenciadora de Unión a CCAAT/metabolismo , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Expresión Génica/efectos de los fármacos , Humanos , Ratones , Obesidad/genética , Obesidad/fisiopatología , PPAR gamma/genética , PPAR gamma/metabolismoRESUMEN
PG201 is an ethanol extract prepared from a specially designed botanical formulation and has previously been shown to contain strong anti-arthritic activities by controlling inflammation and cartilage destruction in two animal models [1,2]. In the present study, we evaluated the effects of PG201 on the expression of heme oxygenase-1 (HO-1). The treatment of Raw264.7 cells (a murine macrophage cell line) and bone marrow-derived macrophages (BMDMs) with PG201 increased the protein and RNA levels of HO-1. The results from a reporter plasmid assay indicated that PG201 induced HO-1 promoter activity through the stress response element present in the two enhancers of the HO-1 promoter. The treatment of cells with PG201 increased the total amount and the nuclear level of NF-E2-related factor 2 (Nrf2). Protein analysis using BMDMs from Nrf2 knockout mice showed that Nrf2 was necessary for the PG201-mediated induction of HO-1 expression. The PG201-mediated induction of these anti-oxidative stress factors was inhibited by a specific inhibitor of phosphatidylinositol 3-kinase (PI3K), but not by inhibitors of p38, ERK and JNK mitogen-activated protein kinases. Furthermore, the results from an experiment involving a specific siRNA and chemical inhibitors for HO-1 showed that the PG201-mediated increase of the HO-1 protein contributed to the suppression of inducible nitric oxide synthase (iNOS) and nitrite production stimulated by lipopolysaccharide. Taken together, these results suggest that PG201 activates Nrf2 through the PI3K signal transduction pathway, increases the expression of HO-1, and subsequently decreases the production of iNOS and nitrite, eventually exerting anti-inflammatory activities.
Asunto(s)
Hemo-Oxigenasa 1/biosíntesis , Factor 2 Relacionado con NF-E2/metabolismo , Nitritos/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Extractos Vegetales/farmacología , Análisis de Varianza , Animales , Línea Celular , Regulación hacia Abajo/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/metabolismo , Ratones , Ratones Noqueados , Factor 2 Relacionado con NF-E2/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Regiones Promotoras Genéticas/efectos de los fármacos , Transducción de Señal/efectos de los fármacosRESUMEN
1-Heteroaryl-2-aryl-1H-benzimidazole derivatives were synthesized as inhibitors of c-Jun N-terminal kinases, JNK3. Their activities were evaluated through measurement of Kd using SPR, JNK3 kinase assay, and cell-viability of human neuroblastoma cells. Most tested compounds showed high affinity (10 µM-46 nM) to JNK3. Among them, compound 16f exhibited potent activities (Kd=46 nM). Especially, 16f was also found to present a potent cell protective effect (IC50=1.09 µM) against toxicity induced by anisomycin, showing a possibility as protective therapeutics in neuronal cell apoptosis.