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MicroRNAs (miRNAs) are a family of non-coding RNAs that play crucial roles in regulating various normal cellular responses. Recent studies revealed that the canonical miRNA biogenesis pathway is subject to sophisticated regulation. Hormonal control of miRNA biogenesis by androgen and estrogen has been demonstrated, but the direct effects of the glucocorticoid receptor (GR) on miRNA biogenesis are unknown. This study revealed the role of GR in miRNA maturation. We showed that two GR agonists, dexamethasone and ginsenoside-Rg1 rapidly suppressed the expression of mature miR-15b, miR-23a, and miR-214 in human endothelial cells. RNA pulldown coupled with proteomic analysis identified GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) as one of the RNA-binding proteins mediating GR-regulated miRNA maturation. Activated GR induced phosphorylation of v-AKT Murine Thymoma Viral Oncogene Homologue (AKT) kinase, which in turn phosphorylated and promoted nuclear translocation of G3BP1. The nuclear G3BP1 bound to the G3BP1 consensus sequence located on primary miR-15b~16-2 and miR-23a~27a~24-2 to inhibit their maturation. The findings from this study have advanced our understanding of the non-genomic effects of GR in the vascular system.
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ADN Helicasas/metabolismo , MicroARNs/genética , Proteínas de Unión a Poli-ADP-Ribosa/metabolismo , ARN Helicasas/metabolismo , Proteínas con Motivos de Reconocimiento de ARN/metabolismo , Receptores de Glucocorticoides/metabolismo , Transporte Activo de Núcleo Celular , Secuencia de Bases , Sitios de Unión , ADN Helicasas/química , Células Endoteliales , Regulación de la Expresión Génica/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana , Humanos , MicroARNs/química , MicroARNs/metabolismo , Proteínas de Unión a Poli-ADP-Ribosa/química , Unión Proteica/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Helicasas/química , Proteínas con Motivos de Reconocimiento de ARN/química , Receptores de Glucocorticoides/agonistasRESUMEN
Therapeutic angiogenesis has been implicated in ischemic diseases and wound healing. Ginsenoside-Rg1 (Rg1), one of the most abundant active components of ginseng, has been demonstrated as an angiogenesis-stimulating compound in different models. There is increasing evidence implicating microRNAs (miRNAs), a group of non-coding RNAs, as important regulators of angiogenesis, but the role of microRNAs in Rg1-induced angiogenesis has not been fully explored. In this report, we found that stimulating endothelial cells with Rg1 could reduce miR-23a expression. In silico experiments predicted hepatocyte growth factor receptor (MET), a well-established mediator of angiogenesis, as the target of miR-23a. Transfection of the miR-23a precursor or inhibitor oligonucleotides validated the inverse relationship of miR-23a and MET expression. Luciferase reporter assays further confirmed the interaction between miR-23a and the MET mRNA 3'-UTR. Intriguingly, ginsenoside-Rg1 was found to increase MET protein expression in a time-dependent manner. We further demonstrated that ginsenoside-Rg1-induced angiogenic activities were indeed mediated through the down-regulation of miR-23a and subsequent up-regulation of MET protein expression, as confirmed by gain- and loss-of-function angiogenic experiments. In summary, our results demonstrated that ginsenoside-Rg1 could induce angiogenesis by the inverse regulation of MET tyrosine kinase receptor expression through miR-23a. This study has broadened our understanding of the non-genomic effects of ginsenoside-Rg1, and provided molecular evidence that warrant further development of natural compound as novel angiogenesis-promoting therapy.
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Inductores de la Angiogénesis/farmacología , Ginsenósidos/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , MicroARNs/metabolismo , Neovascularización Fisiológica/efectos de los fármacos , Proteínas Proto-Oncogénicas c-met/metabolismo , Regiones no Traducidas 3' , Sitios de Unión , Células Cultivadas , Relación Dosis-Respuesta a Droga , Regulación de la Expresión Génica , Células Endoteliales de la Vena Umbilical Humana/enzimología , Humanos , MicroARNs/genética , Proteínas Proto-Oncogénicas c-met/genética , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , TransfecciónRESUMEN
Ginsenoside Rb1 (Rb1) is the most predominant ginsenoside isolated from the roots of ginseng (Panax ginseng C. A. Meyer). This compound is active in various human biological pathways that are involved in human collagen synthesis and inhibition of cell apoptosis. In this study, the skin-whitening effects of Rb1 were investigated in B16 melanoma cells. Our results showed that Rb1 inhibited melanogenesis in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16 cells in a dose-dependent manner, which collectively indicated that Rb1 may have skin-whitening effects and may be formulated into skin-whitening products for skin care. Accordingly, a ginsenoside collagen transdermal patch was developed as a vehicle to topically deliver Rb1 into pig skin. The percutaneous permeation, retention within skin, and release in vitro of Rb1 from seven transdermal patch formulas were studied. It was determined that the best formula for ginsenoside collagen transdermal patch is made of protein collagen hydrolysate powder (PCHP) 2.0% (w/w), methyl cellulose (MC) 0.5% (w/w), polyethyleneglycol 6000 (PEG6000) 0.5% (w/w), ginsenoside 0.036% (w/w), azone 0.4% (v/w), menthol 0.20% (w/w), and water.
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Ginsenósidos/uso terapéutico , Hiperpigmentación/tratamiento farmacológico , Melaninas/antagonistas & inhibidores , Melaninas/biosíntesis , Administración Cutánea , Animales , Línea Celular Tumoral , Supervivencia Celular , Colágeno , Preparaciones de Acción Retardada , Ginsenósidos/administración & dosificación , Humanos , Técnicas In Vitro , Melanoma Experimental/tratamiento farmacológico , Melanoma Experimental/metabolismo , Monofenol Monooxigenasa/metabolismo , Permeabilidad , Porcinos , Parche TransdérmicoRESUMEN
Introduction: Pathological angiogenesis, the abnormal or excessive generation of blood vessels, plays an important role in many diseases including cancer, diabetic retinopathy, psoriasis, and arthritis. Additionally, increasing evidence supports the close linkage between angiogenesis and inflammation. Snake venoms are a rich natural source of biologically active molecules and carry rich potential for the discovery of anti-angiogenic and anti-inflammatory modulators. Methods: Here, we isolated and purified a novel protein, ZK002, from the venom of the snake Deinagkistrodon acutus, and investigated its anti-angiogenic and anti-inflammatory activities and mechanisms. Results: ZK002 was identified as a 30 kDa heterodimeric protein of α and ß chains, which exhibited anti-angiogenic activity in various in vitro assays. Mechanistically, ZK002 inhibited activation of VEGF signaling and related mediators including eNOS, p38, LIMK, and HSP27. ZK002 also upregulated the metalloproteinase inhibitor TIMP3 and inhibited components of the VEGF-induced signaling cascade, PPP3R2 and SH2D2A. The anti-angiogenic activity of ZK002 was confirmed in multiple in vivo models. ZK002 could also inhibit the in vitro expression of pro-inflammatory cytokines, as well as in vivo inflammation in the carrageenin-induced edema rat model. Conclusion: Our findings highlight the potential for further development of ZK002 as a dual function therapeutic against diseases with involvement of pathogenic angiogenesis and chronic inflammation.
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Antrodia camphorata is an extremely rare fungus native to the forested regions of Taiwan. It is also a traditional Chinese medicine, and Taiwanese aborigines applied it for treating liver diseases and protecting from food and drug intoxication. Scientific studies have demonstrated that A. camphorata crude extracts and pure compounds possess a variety of beneficial functions, such as anti-hypertensive, anti-hyperlipidemic, anti-inflammatory, anti-oxidant, anti-tumor, and immuno-modulatory activities. Recent studies have shown that many of these biological and pharmacological activities can be attributed to various active constituents, including polysaccharides, terpenoids, steroids, lignans, benzoquinone derivatives, benzenoids, and maleic and succinic acid derivatives. A. camphorata has been considered as a novel phytotherapeutic agent. However, detailed mechanistic studies or even clinical trials on A. camphorata are still rare. With the help of modern analytical techniques, it is not surprising that many novel constituents are being identified or fractionated from A. camphorata mycelium and fruiting bodies. This review summarizes the latest published results from A. camphorata research, focusing on the biological and pharmacological activities of the crude extract and known constituents of A. camphorata.
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Antrodia/metabolismo , Factores Biológicos/farmacología , Medicina Tradicional China , Antiinflamatorios/química , Antiinflamatorios/farmacología , Antrodia/química , Factores Biológicos/química , Factores Inmunológicos/química , Factores Inmunológicos/farmacología , TaiwánRESUMEN
The objective of the work was to study behavioural change of Cyclope neritea (sea snail) and Nassarius mutabilis (land snail) upon exposure to different levels of PAHs. Snail's behaviour was translated and expressed in Behavioural State Score (BSS) where the score ranged from "0" to "5" points refers to the ascending level of locomotion of a snail. A significant difference was found in snail's behaviour in 25.0 mg/L than in 0.5 mg/L with p value smaller than 0.01. BSS scores appear most frequent on the treatment and control group were 5 (61.5-64.5%) and 2 (41.0-45.0%), respectively. Intersex behaviour was found in all species (i.e. the same sex was grouped together) regardless of PAH concentrations. This is the first reported to study the behavioural change of snail sampled in Hong Kong area when exposed to PAHs. Further studies should be carried on the impact of snail's behaviour exposure on each congener in the family of PAHs.
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Conducta Animal/efectos de los fármacos , Contaminantes Ambientales/toxicidad , Hidrocarburos Policíclicos Aromáticos/toxicidad , Caracoles/fisiología , Animales , Monitoreo del Ambiente , Hong KongRESUMEN
The objective of this study is to investigate the contributions of Mikania micrantha (chinese creeper) to remediate heavy metal pollutants present in an e-waste-contaminated soil. Different proportions of e-waste soil (0%, 25%, 50%, 75%, and 100% w/w) planted with Cynodon dactylon together with the test species were prepared for testing relative seed germination (RSG), relative root growth (RRG), and germination index (GI) tests. A significant higher value of GI (77.8%) was found in M. micrantha than that of the other species when planting in 100% e-waste-contaminated soil. A significant correlation (< 0.05) was found between heavy metal concentration and germination assays in M. micrantha. A significant decrease in heavy metal concentration of the polluted soil after the experiment indicated that biomolecule development studies to determine the aggregate benefit of M. micrantha for phytoremediation remain to be studied in future.
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Residuos Electrónicos , Metales Pesados/metabolismo , Mikania/metabolismo , Contaminantes del Suelo/metabolismo , Biodegradación Ambiental , Germinación , Metales Pesados/análisis , Suelo , Contaminantes del Suelo/análisisRESUMEN
The cell migration/wounding assay is a commonly used method to study cell migration and other biological processes, such as angiogenesis and tumor metastasis. In this assay, cells are grown to form a confluent monolayer and a mechanical wound is created by scratching with a device. Then the migration rate of the cells towards the denuded area can be monitored by imaging. Our 8-channel mechanical wounder is designed to tackle most of the problems associated with the cell migration assay. Firstly, our wounder can be easily sterilized by autoclaving or with common disinfectants. Secondly, the individual adjustable pins allow even contact with the cell culture plate so that sharp and reproducible wounds can be created. Thirdly, the guiding bars on both sides of the wounder ensure consistent wounding position in each well. The use of disposable plastic pipette tips for wounding can further provide better handling of the wounder as well as to minimize cross-contamination. In conclusion, our cell wounder can provide researchers with a user friendly and reproducible device for performing the cell migration assay using the standard 96-well culture plate.
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Ensayos de Migración Celular/instrumentación , Movimiento Celular/fisiología , Cicatrización de Heridas , Células Cultivadas , HumanosRESUMEN
BACKGROUND: Our previous studies have demonstrated that ginsenoside-Rg1 can promote angiogenesis in vitro and in vivo through activation of the glucocorticoid receptor (GR). Furthermore, microRNA (miRNA) expression profiling has shown that Rg1 can modulate the expression of a subset of miRNAs to induce angiogenesis. Moreover, Rb1 was shown to be antiangiogenic through activation of a different pathway. These studies highlight the important functions of miRNAs on ginseng-regulated physiological processes. The aim of this study was to determine the angiogenic properties of Korean Red Ginseng extract (KGE). METHODS AND RESULTS: Combining in vitro and in vivo data, KGE at 500 µg/mL was found to induce angiogenesis. According to the miRNA sequencing, 484 differentially expressed miRNAs were found to be affected by KGE. Among them, angiogenic-related miRNAs; miR-15b, -23a, -214, and -377 were suppressed by KGE. Meanwhile, their corresponding angiogenic proteins were stimulated, including vascular endothelial growth factor, vascular endothelial growth factor receptor-2, endothelial nitric oxide synthase, and MET transmembrane tyrosine kinase. The miRNAs-regulated signaling pathways of KGE were then found by Cignal 45-Pathway Reporter Array, proving that KGE could activate GR. CONCLUSION: KGE was found capable of inducing angiogenesis both in vivo and in vitro models through activating GR. This study provides a valuable insight into the angiogenic mechanisms depicted by KGE in relation to specific miRNAs.
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MicroRNAs (miRNAs) are small non-coding RNAs that play a crucial role in pathogenesis of human cancers. Several miRNAs have been shown to involve in nasopharyngeal carcinoma (NPC) pathogenesis through alteration of gene networks. A global view of the miRNA expression profile of clinical specimens would be the best way to screen out the possible miRNA candidates that may be involved in disease pathogenesis. In this study, we investigated the expression profiles of miRNA in formalin-fixed paraffin-embedded tissues from patients with undifferentiated NPC versus non-NPC controls using a miRNA real-time PCR platform, which covered a total of 95 cancer-related miRNAs. Hierarchical cluster analysis revealed that NPC and non-NPC controls were clearly segregated. Promisingly, 10 miRNA candidates were differentially expressed. Among them, 9 miRNAs were significantly up-regulated of which miR-205 and miR-196a showed the most up-regulated in NPC with the highest incidence percentage of 94.1% and 88.2%, respectively, while the unique down-regulated miR-150 was further validated in patient sera. Finally, the in vitro gain-of-function and loss-of-function assays revealed that miR-150 can modulate the epithelial-mesenchymal-transition property in NPC/HK-1 cells and led to the cell motility and invasion. miR-150 may be a potential biomarker for NPC and plays a critical role in NPC tumourigenesis.
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Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Carcinoma Nasofaríngeo/genética , Neoplasias Nasofaríngeas/genética , Adulto , Anciano , Línea Celular Tumoral , Movimiento Celular/genética , Transición Epitelial-Mesenquimal/genética , Femenino , Humanos , Masculino , Persona de Mediana Edad , Carcinoma Nasofaríngeo/patología , Neoplasias Nasofaríngeas/patología , Metástasis de la NeoplasiaRESUMEN
Euxanthone, a neuritogenic agent isolated from the medicinal herb Polygala caudata, has been shown to induce morphological differentiation and neurite outgrowth in murine neuroblastoma Neuro 2a cells (BU-1 subclone). In order to elucidate the underlying mechanisms of euxanthone-induced neurite outgrowth, a proteomic approach was employed. In the present study, two dimensional (2-D) gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight (MALDI-ToF) mass spectrometry were performed to investigate the alterations in protein expression profile of euxanthone-treated BU-1 cells. Fourteen identified proteins were changed in expression levels after induction of neurite growth. These proteins included participants in transcription and cell cycle regulation, calcium influx and calcium signaling, fatty acid metabolism, cytoskeleton reorganization, casein kinase signal transduction, putative transbilayer amphipath transport and protein biosynthesis. Among the 14 identified proteins, E2F transcription factor 5 (E2F-5) was significantly up-regulated after euxanthone treatment. Go6976, a protein kinase C (PKC) alpha/betaI inhibitor, was found to inhibit neuritogenesis and expression of E2F-5 in the euxanthone-treated BU-1 cells, while SH-6, the Akt/PKB inhibitor, had no inhibitory effect. The gene silencing of E2F-5 by small interfering RNA (siRNA) was found to abolish the euxanthone-induced neurite outgrowth. In conclusion, these results indicated that the transcription factor E2F-5 was actively involved in the regulation of euxanthone-induced neurite outgrowth via PKC pathway.
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Diferenciación Celular/efectos de los fármacos , Factores de Transcripción E2F/metabolismo , Neuritas/metabolismo , Proteína Quinasa C/metabolismo , Xantonas/farmacología , Animales , Carbazoles/farmacología , Línea Celular Tumoral , Factores de Transcripción E2F/genética , Electroforesis en Gel Bidimensional , Silenciador del Gen , Indoles/farmacología , Neuritas/efectos de los fármacos , Neuroblastoma/metabolismo , Neuroblastoma/patología , Proteína Quinasa C/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Proteómica/métodos , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
BACKGROUND: Ginsenoside-Rg3, the pharmacologically active component of red ginseng, has been found to inhibit tumor growth, invasion, metastasis, and angiogenesis in various cancer models. Previously, we found that 20(R)-ginsenoside-Rg3 (Rg3) could inhibit angiogenesis. Since microRNAs (miRNAs) have been shown to affect many biological processes, they might play an important role in ginsenoside-mediated angiomodulation. METHODS: In this study, we examined the underlying mechanisms of Rg3-induced angiosuppression through modulating the miRNA expression. In the miRNA-expression profiling analysis, six miRNAs and three miRNAs were found to be up- or down-regulated in vascular-endothelial-growth-factor-induced human-umbilical-vein endothelial cells (HUVECs) after Rg3 treatment, respectively. RESULTS: A computational prediction suggested that mature hsa-miR-520h (miR-520h) targets ephrin receptor (Eph) B2 and EphB4, and hence, affecting angiogenesis. The up-regulation of miR-520h after Rg3 treatment was validated by quantitative real-time polymerase chain reaction, while the protein expressions of EphB2 and EphB4 were found to decrease, respectively. The mimics and inhibitors of miR-520h were transfected into HUVECs and injected into zebra-fish embryos. The results showed that overexpression of miR-520h could significantly suppress the EphB2 and EphB4 protein expression, proliferation, and tubulogenesis of HUVECs, and the subintestinal-vessel formation of the zebra fish. CONCLUSION: These results might provide further information on the mechanism of Rg3-induced angiosuppression and the involvement of miRNAs in angiogenesis.
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Artificial sweeteners have been recognized as emerging contaminants due to their wide application, environmental persistence and ubiquitous occurrence. Among them, acesulfame has attracted much attention. After being discharged into the environment, acesulfame undergoes photolysis naturally. However, acesulfame photodegradation behavior and identity of its transformation products, critical to understanding acesulfame's environmental impact, have not been thoroughly investigated. The present study aimed to fill this knowledge gap by a laboratory simulation study in examining acesulfame transformation products and pathways under UV-C photolysis in the presence of TiO2. Photodegradation products of acesulfame were isolated and analyzed using the LC-IM-QTOF-MS coupled with LC Ion Trap MS in the MS(n) mode. Our results show six new transformation products that have not been previously identified. The molecular structures and transformation pathways were proposed. Further embryotoxicity tests showed that acesulfame transformation products at the low g L(-1) level produced significant adverse effects in tail detachment, heart rate, hatching rate and survival rate during fish embryo development. The identification of additional transformation products with proposed transformation pathways of acesulfame, the increased toxicity of acesulfame after photolysis, and the fact that the accumulation of acesulfame transformation products is increasingly likely make acesulfame contamination even more important. Water resource control agencies need to consider legislation regarding acesulfame and other artificial sweeteners, while further studies are carried out, in order to protect the safety of this most vital resource.
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Embrión no Mamífero , Fotólisis , Tiazinas/química , Tiazinas/toxicidad , Pez Cebra/embriología , Animales , Cromatografía Liquida , Espectrometría de Masas , Edulcorantes/química , Edulcorantes/toxicidad , Titanio/química , Pruebas de Toxicidad Aguda , Rayos Ultravioleta , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/toxicidadRESUMEN
Methyl protodioscin is a furostanol bisglycoside with antitumor properties. The present study investigated its effects on human chronic myelogenous leukemia K562 cells. Cell cycle analysis showed that methyl protodioscin caused distinct G2/M arrest, with the appearance of polyploidy population. The levels of cyclin B1 decreased, whereas Cdc2 kept at a steady level. Subsequent apoptosis after G2/M blockage was demonstrated through DNA fragmentation and the annexin V staining assay. Methyl protodioscin induced a biphasic alteration (i.e. an early hyperpolarization, followed by depolarization) in mitochondrial membrane potential of K562 cells. The transient decline of intracellular Ca2+ concentration was observed at early stage. The generation of reactive oxygen species was also detected. The anti-apoptotic Bcl-x(L) transiently increased and then decreased. And the pro-apoptotic Bax was markedly up-regulated. Taken together, these data demonstrated that methyl protodioscin inhibits K562 cell proliferation via G2/M arrest and apoptosis, with mitochondrial hyperpolarization and the disruption of Ca2+ homeostasis playing important roles.
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Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Diosgenina/análogos & derivados , Diosgenina/farmacología , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Saponinas/farmacología , Calcio/farmacocinética , Proliferación Celular , Daño del ADN , Homeostasis , Humanos , Mitocondrias/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Células Tumorales Cultivadas , Regulación hacia Arriba , Proteína X Asociada a bcl-2RESUMEN
BACKGROUND: Nasopharyngeal carcinoma (NPC) has a high incidence rate in Southern China. Although there are conventional therapies, the side effects and toxicities are not always tolerable for patients. Recently, the tumoricidal effect of ginsenosides on different cancer cells has been studied. This study aims to investigate the anti-cancer effect of ginsenosides on NPC cells and their underlying mechanism. METHODS: The cytotoxicity of ginsenosides on NPC cell line HK-1 was measured by MTT assay. Apoptosis was detected by propidium iodide staining followed by flow cytometry. A xenograft tumor model was established by injecting nude mice with HK-1 cells. The activation of caspases and apoptosis-inducing factor (AIF) were evaluated by Western blot analysis. Nuclear translocation of AIF was also studied by immunofluorescence staining. Mitochondrial membrane potential was measured by JC-1 dye using flow cytometry. RESULTS: Four ginsenosides, 20 (S)-Rh2, compound K (CK), panaxadiol (PD) and protopanaxadiol (PPD), induced apoptotic cell death in HK-1 cells in a concentration-dependent manner. CK inhibited HK-1 xenograft tumor growth most extensively and depleted mitochondrial membrane potential depolarization and induced translocation of AIF from cytoplasm to nucleus in HK-1 cells. In addition, depletion of AIF by siRNA abolished CK-induced HK-1 cell death. CONCLUSION: Ginsenoside CK-induced apoptosis of HK-1 cells was mediated by the mitochondrial pathway and could significantly inhibit tumor growth in vivo.
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Antrodia cinnamomea (AC) is an endemic mushroom species of Taiwan, and has been demonstrated to possess diverse biological and pharmacological activities, such as anti-hypertension, anti-hyperlipidemia, anti-inflammation, anti-oxidation, anti-tumor, and immunomodulation. This review focuses on the inhibitory effects of AC on hepatitis, hepatocarcinoma, and alcohol-induced liver diseases (e.g., fatty liver, fibrosis). The relevant biochemical and molecular mechanisms are addressed. Overall, this review summarizes the hepatoprotective activities in vitro and in vivo. However, there is no doubt that human and clinical trials are still limited, and further studies are required for the development of AC-related products.
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Tumor necrosis factor-alpha (TNF- α ) was reported as anticancer therapy due to its cytotoxic effect against an array of tumor cells. However, its undesirable responses of TNF- α on activating NF- κ B signaling and pro-metastatic property limit its clinical application in treating cancers. Therefore, sensitizing agents capable of overcoming this undesirable effect must be valuable for facilitating the usage of TNF- α -mediated apoptosis therapy for cancer patients. Previously, saikosaponin-d (Ssd), a triterpene saponin derived from the medicinal plant, Bupleurum falcatum L. (Umbelliferae), showed to exhibit a variety of pharmacological activities such as antiinflammation, antibacteria, antivirus and anticancer. Recently, we found that Ssd could inhibit the activated T lymphocytes via suppression of NF- κ B, NF-AT and AP-1 signaling. Here, we showed that Ssd significantly potentiated TNF- α -mediated cell death in HeLa and HepG2 cancer cells via suppression of TNF- α -induced NF- κ B activation and its target genes expression involving cancer cell proliferation, invasion, angiogenesis and survival. Also, Ssd revealed a significant potency of abolishing TNF- α -induced cancer cell invasion and angiogenesis in HUVECs while inducing apoptosis via enhancing the loss of mitochondrial membrane potential in HeLa cells. Collectively, these findings indicate that Ssd has a significant potential to be developed as a combined adjuvant remedy with TNF- α for cancer patients.
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Wrinkle formation is one of the primary characteristics of skin aging, the major cause of wrinkle is the loss of structural protein type I collagen in dermal layer of skin. Topical application of natural substances to reduce wrinkle is gaining attention in recent years. Although a number of polyphenoic compounds are suggested to prevent ultraviolet-induced wrinkle, very few of them are able to increase type I collagen synthesis directly. Ginseng has been known in folk medicine of its beneficial effect to skin. The present study investigate the effect of ginsenoside on type I collagen induction in human dermal fibroblasts. Ginsenoside Rb1 was shown to induce type I collagen expression in dermal fibroblasts in a dose- and time-dependent manner. Recent studies suggest the important post-transcriptional regulatory role of microRNAs; here we demonstrated that miR-25 can directly inhibit type I collagen protein expression, and treatment of fibroblasts with Rb1 can reduce the inhibition by decreasing miR-25 level. Furthermore, we identified that the nuclear receptor, peroxisome proliferator-activated receptor-delta (PPARδ) is the key mediator of Rb1-induced type I collagen expression. Knockdown of PPARδ by small-interference RNA abolished the Rb1-induced type I collagen production and reversed the Rb1-suppressed miR-25 expression. These results demonstrated that ginsenoside Rb1 can increase target gene expression through transcriptional pathway, at the same time, inhibit the corresponding miRNA expression to minimize the translation repression. Furthermore, this study provide solid support of ginsenoside Rb1-induced type I collagen expression, which warrant further study in the dermatological application of ginsenosides in skin disorders.
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Colágeno Tipo I/biosíntesis , Dermis/citología , Fibroblastos/efectos de los fármacos , Ginsenósidos/farmacología , PPAR delta/metabolismo , Células Cultivadas , Colágeno Tipo I/genética , Relación Dosis-Respuesta a Droga , Fibroblastos/metabolismo , Técnicas de Silenciamiento del Gen , Humanos , MicroARNs/genética , MicroARNs/metabolismo , PPAR delta/genética , ARN Interferente Pequeño/genéticaRESUMEN
Angiogenesis, the development of neovessels from pre-existing vessels, is obligatory for solid tumors survival, growth, invasion, and metastasis. Many anti-angiogenic agents are small molecules originated from natural sources. Recently, angiosuppressive effects of indirubin and its derivatives, the active components in indigo-producing herbs, have been shown to possess anti-viral and anti-inflammatory potentials. In this study, we identified another indirubin derivative, indirubin-3'-(2,3 dihydroxypropyl)-oximether (E804), could exhibit potent angiosuppressive effects. In vitro study showed that E804 could significantly inhibit human umbilical vein endothelial cells proliferation, migration, and tube formation in a concentration-dependent manner (0.4-40 µM); at the concentration of 1 µmol or above, angiosuppressive potency of E804 was found to be more significant than indirubin-3'-oxime. Using in vivo Matrigel plug model and directed-in vivo-angiogenesis-assay (DIVAA), E804 was shown more effective to attenuate the VEGF/bFGF-induced neovessel formation. The hemoglobin content and the invaded endothelial cells in the implants were also greatly reduced. Results from the aortic ring assay indicated E804 (4 µM) could completely suppress ex vivo sprouting of endothelial cells from the rat aorta fragments; with concomitant reduction of gelatinolytic activities of matrix metalloproteinase-2 and -9. E804 also concentration-dependently (0.04-10 µM) inhibited the subintestinal vessels formation in zebrafish embryos. This study provides the first evidence that E804, a novel indirubin derivative, could more effectively inhibit angiogenesis. With the improved anti-angiogenic potency when compared with indirubin-3'oxime, E804 would be a new potential candidate in the treatment of angiogenesis-dependent diseases.
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Indoles/farmacología , Neovascularización Patológica/tratamiento farmacológico , Animales , Aorta/citología , Células Cultivadas , Embrión no Mamífero/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Células Endoteliales/enzimología , Humanos , Indoles/química , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Estructura Molecular , Oximas , Ratas , Pez CebraRESUMEN
Ginsenosides are considered the major constituents that are responsible for most of the pharmacological actions of ginseng. However, some ginsenosides exist as stereoisomeric pairs, detailed and molecular exposition based on the structural differences of ginsenoside stereoisomers has not been emphasized in most studies. Here we explore the functional differences of ginsenoside Rg3 stereoisomers on angiogenesis. In this study, we demonstrated the distinctive differential angiogenic activities of 20(S)-Rg3 and 20(R)-Rg3 stereoisomers. 20(S)-Rg3 at micromolar concentration promotes human endothelial cells proliferation, migration and tube formation in vitro, as well as ex vivo endothelial sprouting. The effects induced by 20(S)-Rg3 are significantly more potent than 20(R)-Rg3. These effects are partially mediated through the activation of AKT/ERK-eNOS signaling pathways. Moreover, knockdown of peroxisome proliferator-activated receptor-gamma (PPARγ) by specific small interference RNA abolished the 20(S)-Rg3-induced angiogenesis, indicating that PPARγ is responsible for mediating the angiogenic activity of Rg3. Using reporter gene assay, the PPARγ agonist activity of 20(S)-Rg3 has been found 10-fold higher than that of 20(R)-Rg3. Computer modeling also revealed the differential binding is due to the chiral center of 20(S)-Rg3 can form a critical hydrogen bond with Tyr473 of PPARγ ligand binding domain. The present study elucidated the differential angiogenic effects of Rg3 stereoisomers by acting as agonist of PPARγ. The results shed light on the structural difference between two ginsenoside stereoisomers that can lead to significant differential physiological outcomes which should be carefully considered in the future development of ginsenoside-based therapeutics.