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1.
Med Sci Monit ; 26: e920557, 2020 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-32186283

RESUMO

BACKGROUND Doxorubicin-induced myocardial toxicity is associated with oxidative stress, cardiomyocyte, apoptosis, and loss of contractile function. Previous studies showed that microRNA-375 (miR-375) expression was increased in mouse models of heart failure and clinically, and that inhibition of miR-375 reduced inflammation and increased survival of cardiomyocytes. This study aimed to investigate the effects and mechanisms of inhibition of miR-375 in a mouse model of doxorubicin-induced cardiac toxicity in vivo and in doxorubicin-treated rat and mouse cardiomyocytes in vitro. MATERIAL AND METHODS The mouse model of doxorubicin-induced cardiac toxicity was developed using an intraperitoneal injection of doxorubicin (15 mg/kg diluted in 0.9% saline) for eight days. Treatment was followed by a single subcutaneous injection of miR-375 inhibitor. H9c2 rat cardiac myocytes and adult murine cardiomyocytes (AMCs) were cultured in vitro and treated with doxorubicin, with and without pretreatment with miR-375 inhibitor. RESULTS Doxorubicin significantly upregulated miR-375 expression in vitro and in vivo, and inhibition of miR-375 re-established myocardial redox homeostasis, prevented doxorubicin-induced oxidative stress and cardiomyocyte apoptosis, and activated the PDK1/AKT axis by reducing the direct binding of miR-375 to 3' UTR of the PDK1 gene. Inhibition of PDK1 and AKT abolished the protective role of miR-375 inhibition on doxorubicin-induced oxidative damage. CONCLUSIONS Inhibition of miR-375 prevented oxidative damage in a mouse model of doxorubicin-induced cardiac toxicity in vivo and in doxorubicin-treated rat and mouse cardiomyocytes in vitro through the PDK1/AKT signaling pathway.


Assuntos
Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , MicroRNAs/metabolismo , Miócitos Cardíacos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Animais , Apoptose , Cardiotoxicidade , Células Cultivadas , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/efeitos dos fármacos , MicroRNAs/genética , Ratos
2.
Adv Clin Exp Med ; 29(1): 135-145, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32011832

RESUMO

BACKGROUND: Anesthetics, such as isoflurane, sevoflurane, ketamine, and desflurane, are commonly used in clinics. Specifically, isoflurane is one of the most commonly used inhalational anesthetics, which can be used in surgery patients of all ages, including children. OBJECTIVES: The aim of the study was to investigate the mechanisms of vitexin against isoflurane-induced neurotoxicity. MATERIAL AND METHODS: Reference memory testing was performed for 5 days (4 trials, 2 per day) before anesthesia. Reversal testing was performed on the 3rd day after anesthesia. The cell viability and apoptosis of PC-12 cells were detected using MTT and TUNEL assays, respectively. Enzyme-linked immunosorbent assay (ELISA) kits were used to measure serum tumor necrosis factor α (TNF­α), interleukin 6 (IL­6), glutathione (GSH), and superoxide dismutase (SOD) concentrations. The concentration of reactive oxygen species (ROS) was detected using ROS measurement. Expression of miR-409 was determined using quantitative reverse-transcription polymerase chain reaction (qPT-PCR). Protein expression levels were detected using western blotting. RESULTS: Rats treated with isoflurane showed significant increases in the escape latency periods (ELP) and the apoptosis of hippocampus neuron cells; this effect was reversed by 3 mg/kg or 10 mg/kg of vitexin (p < 0.05). Further testing showed that isoflurane could significantly decrease the cell viability and increase the apoptosis of PC-12, the expression of inflammatory cytokines (TNF­α and IL­6) and ROS (p < 0.05). However, these results were reversed by 10/100 µM of vitexin. In addition, vitexin could significantly increase the expression of miR-409 (p < 0.05). Further studies showed that overexpression of miR-409 could significantly promote the effect of vitexin on isoflurane-induced neurotoxicity (p < 0.05). Finally, overexpression miR-409 could significantly increase the expression of p-AMPK/t-AMPK and p-GSK3ß/t-GSK3ß. CONCLUSIONS: Vitexin has protective effects against isoflurane-induced neurotoxicity by targeting miR-409 and the AMPK/GSK3ß pathway.


Assuntos
Anestésicos Inalatórios , Apigenina/farmacologia , Isoflurano/farmacologia , MicroRNAs/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fármacos Neuroprotetores/farmacologia , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP , Animais , Apoptose/efeitos dos fármacos , Criança , Ensaio de Imunoadsorção Enzimática , Glutationa/sangue , Glicogênio Sintase Quinase 3 beta , Humanos , Interleucina-6/sangue , MicroRNAs/genética , Ratos , Espécies Reativas de Oxigênio , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Superóxido Dismutase/sangue , Fator de Necrose Tumoral alfa/sangue
3.
J Appl Oral Sci ; 28: e20190382, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049136

RESUMO

OBJECTIVE: This study aimed to investigate the effects of Maras powder (a type of smokeless tobacco obtained from Nicotiana rustica Linn and mixed with the ashes of wood, especially from oak, walnut or grapevine) on the microRNA (miRNA) deregulation of oral mucosa, and it compares these effects with those of smoking. METHODOLOGY: Oral mucosal samples were collected from 74 patients, consisting of 16 nonusers, 26 smokers, and 32 Maras powder users. Genes associated with oral cancer were selected and 90 microRNAs targeting these genes were identified. MicroRNA were isolated and purified using the microRNA isolation kit. MicroRNA were expressed using Fluidigm RT-PCR. RESULTS: A positive correlation between the duration of Maras powder use with miR-31 expression levels, and a negative correlation between the Maras powder chewing time and miR-372 expression levels was found. In addition, there is a negative correlation between the amount of Maras powder consumed and expression levels of miR-375, miR-378a, miR-145, and miR-10b; moreover, another negative correlation is observed between the number of cigarettes consumed and the expression levels of miR-23a, miR-23b, miR-203a, miR-200b, and miR-375. However, miR-200b and miR-92a levels were downregulated significantly more in Maras powder users when compared with smokers and nonusers (p<0.05). CONCLUSION: The results show both chewing Maras powder and smoking have an effect on deregulation of miR-200b and miR-92a expressions. This leads to the belief that assessing the expression of these two miRNAs is a promising noninvasive method of analysis, especially in mutagen exposures. Finally, large-scale and high-throughput studies may help to identify an extensive miRNA expression profile associated with tobacco use and improve the understanding of oral malignancies.


Assuntos
MicroRNAs/efeitos dos fármacos , Mucosa Bucal/efeitos dos fármacos , Tabaco sem Fumaça/efeitos adversos , Adolescente , Adulto , Idoso , Análise de Variância , Estudos Transversais , Regulação para Baixo , Feminino , Expressão Gênica , Humanos , Masculino , MicroRNAs/análise , Pessoa de Meia-Idade , Neoplasias Bucais/genética , Pós , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Fatores de Tempo , Adulto Jovem
4.
Arch Oral Biol ; 110: 104630, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31837588

RESUMO

OBJECTIVE: This study is to investigate the effect and underlying mechanisms of berberine (BBR) on the proliferation and inflammatory levels of lipopolysaccharide induced human dental pulp fibroblast (LPS-HDPF). METHODS: Different concentrations of LPS were used to induce inflammatory response of HDPF. Cell proliferation was observed in BBR treated HDPF after transfection of miR-21 mimic, miR-21 inhibitor or sh-kBTBD7 plasmids and their negative controls. ELISA was employed to detect the expressions of inflammation related cytokines. Real-time quantitative RT-PCR was applied to estimate the expressions of miR-21 and KBTBD7. KBTBD7 expression was detected by Immunocytochemistry. Western blot analysis showed the protein levels of KBTBD7, Phospho-IKKα/ß, IKKß, Phospho-NF-κB p65, NF-κB p65, and IκBα. Luciferase reporter gene assay was used to determine the interaction between miR-21 and KBTBD7. RESULTS: LPS could promote inflammatory response in HDPF. Down-regulated miR-21 and up-regulated KBTBD7 were found in LPS-HDPF. BBR (25 uM) treatment in LPS-HDPF could ameliorate cell inflammatory response, presented by reduced expressions of IL-1ß, IL-6 and TNF-α, as well as enhanced cell proliferation and miR-21 expression. Moreover, miR-21 negatively targets KBTBD7. Over-expression of miR-21 or silencing of KBTBD7 could enhance the protective role of BBR on LPS-HDPF by inhibiting inflammatory response and promoting cell proliferation. Transfection of miR-21 overexpression or KBTBD7 silencing in BBR treated LPS-HDPF could inhibit activation of NF-κB signal pathway. CONCLUSION: Evidence in this study suggested that BBR mediates LPS induced inflammation in HDPF via miR-21/KBTBD7 axis to regulate NF-κB signal pathway, which may provide theoretical basis for BBR in prevention of pulpitis.


Assuntos
Berberina , Polpa Dentária , MicroRNAs , Berberina/farmacologia , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/metabolismo , Fibroblastos , Humanos , Inflamação , Peptídeos e Proteínas de Sinalização Intracelular , Lipopolissacarídeos , MicroRNAs/efeitos dos fármacos , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais , Transativadores
5.
Int J Mol Sci ; 20(18)2019 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-31540128

RESUMO

Polyphenols are potent micronutrients that can be found in large quantities in various food sources and spices. These compounds, also known as phenolics due to their phenolic structure, play a vital nutrient-based role in the prevention of various diseases such as diabetes, cardiovascular diseases, neurodegenerative diseases, liver disease, and cancers. However, the function of polyphenols in disease prevention and therapy depends on their dietary consumption and biological properties. According to American Cancer Society statistics, there will be an expected rise of 23.6 million new cancer cases by 2030. Due to the severity of the increased risk, it is important to evaluate various preventive measures associated with cancer. Relatively recently, numerous studies have indicated that various dietary polyphenols and phytochemicals possess properties of modifying epigenetic mechanisms that modulate gene expression resulting in regulation of cancer. These polyphenols and phytochemicals, when administrated in a dose-dependent and combinatorial-based manner, can have an enhanced effect on epigenetic changes, which play a crucial role in cancer prevention and therapy. Hence, this review will focus on the mechanisms of combined polyphenols and phytochemicals that can impact various epigenetic modifications such as DNA methylation and histone modifications as well as regulation of non-coding miRNAs expression for treatment and prevention of various types of cancer.


Assuntos
Antineoplásicos/uso terapêutico , Epigênese Genética/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Compostos Fitoquímicos/uso terapêutico , Polifenóis/uso terapêutico , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Antineoplásicos/farmacologia , Metilação de DNA/efeitos dos fármacos , Dieta , Histonas/metabolismo , Humanos , MicroRNAs/efeitos dos fármacos , MicroRNAs/metabolismo , Micronutrientes , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/prevenção & controle , Compostos Fitoquímicos/administração & dosagem , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Polifenóis/administração & dosagem , Polifenóis/química , Polifenóis/farmacologia , Processamento de Proteína Pós-Traducional/efeitos dos fármacos
6.
Medicina (Kaunas) ; 55(9)2019 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-31540331

RESUMO

Background and Objectives: Atropine is a nonselective muscarinic antagonist which has been used to prevent worsening of myopia in children. Different concentrations of atropine were used for myopia, ranging from 0.01% to 1.0%. However, there are still potential toxicity of different doses of atropine to the cornea. Here, we present a study of investigating novel genes potentially involved in the effects of very low dose atropine treatment (0.003%) on corneal epithelial cells using next-generation sequencing (NGS) and bioinformatics approaches. Materials and Methods: Human corneal epithelial cells were treated with 0.003% atropine, cultured until confluence, and RNA extracted for differential expression profiling of mRNA and microRNA (miRNA) between control and atropine-treated corneal epithelial cells. The functional enrichment analysis for differentially expressed genes was performed using two bioinformatics databases, including Database for Annotation, Visualization and Integrated Discovery (DAVID) and Ingenuity® Pathway Analysis (IPA). In addition, potential miRNA-mRNA interactions involved in atropine-treated corneal epithelial cells were predicted and validated using different miRNA target prediction databases. Results: Our results showed 0.003% atropine might suppress the apoptosis of corneal epithelial cells, potentially through Ras and protein kinase A signaling pathways. We also validated the possible miRNA regulations by using TargetScan and miRDB databases. Hsa-miR-651-3p-EPHA7, hsa-miR-3148-TMEM108 and hsa-miR-874-5p-TBX6 were validated as possible miRNA regulations involved in corneal epithelial cells treated with 0.003% atropine. Conclusions: These findings may contribute novel insights into therapeutic strategies for treating cornea with 0.003% atropine.


Assuntos
Atropina/farmacologia , Córnea/citologia , Células Epiteliais/efeitos dos fármacos , MicroRNAs/genética , Antagonistas Muscarínicos/farmacologia , Atropina/uso terapêutico , Biologia Computacional , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , MicroRNAs/efeitos dos fármacos , Antagonistas Muscarínicos/uso terapêutico , Miopia/tratamento farmacológico
7.
Anim Reprod Sci ; 207: 21-35, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31266599

RESUMO

Organotypic culture of testicular fragments from 7-day-old male pigs (Polish White Large) was used. Tissues were treated with an antagonist of G-protein coupled estrogen receptor (GPER) (G-15; 10 nM), and bisphenol A (BPA), and its analogs (TBBPA, TCBPA; 10 nM) alone or in combination and analyzed using electron and light (stainings for collagen fibers, lipid droplet and autophagy markers) microscopes. In addition, mRNA and protein abundances and localization of molecules required for miRNA biogenesis and function (Drosha, Exportin 5; EXPO5, Dicer, and Argonaute 2; AGO2) were assessed together with calcium ion (Ca2+) and estradiol concentrations. Regardless of GPER blockade and/or treatment with BPA, TBBPA and TCBPA, there were no changes in Leydig cell morphology. Also, there were no changes in lipid droplet content and distribution but there were changes in lipid and autophagy protein abundance. In the interstitial tissue, there was an increase of collagen content, especially after treatment with BPA analogs and G-15 + BPA. Independent of the treatment, there was downregulation of EXPO5 and Dicer genes but the Drosha and AGO2 genes were markedly upregulated as a result of treatment with G-15 + BPA and TCBPA, respectively. There was always a lesser abundance of EXPO5 and AGO2 proteins regardless of treatment. There was markedly greater abundances of Drosha after G-15 + BPA treatment, and this also occurred for Dicer after treatment with G-15 + TCBPA. Immunolocalization of miRNA proteins indicated there was a cytoplasmic-nuclear pattern in control and treated cells. There was an increase of Ca2+ concentrations after treatment with G-15 and BPA analogs. Estradiol secretion decreased after antagonist and chemical treatments when these were administered alone, however, there was an increase in estradiol secretion after treatment with combinations of these compounds.


Assuntos
Compostos Benzidrílicos/farmacologia , Epigênese Genética/efeitos dos fármacos , Células Intersticiais do Testículo/efeitos dos fármacos , Fenóis/farmacologia , Receptores Estrogênicos/fisiologia , Receptores Acoplados a Proteínas-G/fisiologia , Testículo/efeitos dos fármacos , Animais , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Interação Gene-Ambiente , Células Intersticiais do Testículo/metabolismo , Masculino , MicroRNAs/efeitos dos fármacos , MicroRNAs/genética , MicroRNAs/metabolismo , Receptores Estrogênicos/genética , Receptores Acoplados a Proteínas-G/genética , Maturidade Sexual/efeitos dos fármacos , Maturidade Sexual/genética , Suínos , Testículo/metabolismo
8.
Artigo em Inglês | MEDLINE | ID: mdl-31263290

RESUMO

Quercetin, a flavonoid with multiple proven health benefits to both man and animals, displays a plethora of biological activities, collectively referred to as pleiotropic. The most studied of these are antioxidant and anti-inflammatory but modulation of signalling pathways is important as well. One of the lesser-known and recently discovered roles of quercetin, is modulation of microRNA (miRNA) expression. miRNAs are important posttranscriptional modulators that play a critical role in health and disease and many of these non-coding oligonucleotides are recognized as oncogenic or tumor suppressor miRNAs. This review is an evaluation of the recent relevant literature on the subject, with focus on the ability of quercetin to modulate miRNA expression. It includes a summary of recent knowledge on miRNAs deregulated by quercetin, an overview of quercetin pharmacokinetics and miRNA biogenesis, for the interested reader.


Assuntos
Expressão Gênica/efeitos dos fármacos , MicroRNAs/efeitos dos fármacos , Quercetina/farmacologia , Animais , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo
9.
Pathol Res Pract ; 215(8): 152494, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31229277

RESUMO

T-cell acute lymphoblastic leukemia (T-ALL) has a relatively improved remission rate, but the poor outcomes are primarily due to resistance and relapse. Moreover, organs infiltration trends to occur during remission. Rapamycin was applied to treat malignancies for decades. In this investigation, we aimed to explore the molecular mechanisms and pathway changes during the T-ALL therapeutic process. T-ALL cell line Molt-4 cells were treated with rapamycin and performed microarray analysis to identify the deregulated miRNAs and mRNAs (log2 fold change>2 or <-2). To obtain regulatory miRNA/mRNA network, miRNA target prediction softwares and Cytoscape were used to plot and modularize the rapamycin treatment-related network. Surprisingly, the enriched pathways were not involved in mediating either cell death or apoptosis but were responsible for angiogenesis, cell survival, and anti-apoptosis, which is consistent with the Gene Ontology analysis and PPI network based on all deregulated mRNAs, indicating that these elements likely play a role in promoting Molt-4 cell survival or escaping from rapamycin. The expression of 3 miRNAs (miR-149-3p, miR-361-3p, and miR-944) and their putative targets, which play central roles in their module, were validated by qRT-PCR. These results provide novel insight into potentially relevant biological pathways for T-ALL cells escaping from chemotherapy or developing central nervous system infiltration.


Assuntos
Linhagem da Célula/efeitos dos fármacos , MicroRNAs/metabolismo , Sirolimo/farmacologia , Linfócitos T/efeitos dos fármacos , Linhagem Celular Tumoral , Perfilação da Expressão Gênica/métodos , Humanos , MicroRNAs/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Leucemia-Linfoma Linfoblástico de Células T Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patologia , RNA Mensageiro/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Linfócitos T/metabolismo
10.
Life Sci ; 231: 116528, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31176784

RESUMO

AIMS: Lycorine is a kind of natural alkaloid with anti-cancer potential. It has been demonstrated that lycorine processes high activity and specificity against the progression of cancers. However, the underlying molecular mechanisms by which lycorine regulates the formation and development of non-small cell lung cancer (NSCLC) remain largely unknown. MAIN METHODS: The effects of lycorine on the growth of NSCLC cells were determined by the cell counting kit-8 (CCK-8) assay, colony formation and flow cytometry analysis. RT-qPCR was performed to detect the expression of microRNA with lycorine treatment. The binding of miRNA and target genes was confirmed by luciferase reporter assay. KEY FINDINGS: Lycorine significantly inhibited the proliferation and induced apoptosis of NSCLC cells. Mechanistically, lycorine up-regulated the expression of microRNA-186 in NSCLC cells. Depletion of miR-186 significantly reversed the suppressive effect of lycorine on the proliferation of NSCLC cells. Furthermore, the cyclin dependent kinase 1 (CDK1) was identified as one of the binding candidates of miR-186. Experimental analysis showed that miR-186 bound the 3'-untranslated region (3'-UTR) of CDK1 and suppressed the level of CDK1 in NSCLC cells. Consistently, exposure of lycorine significantly decreased the expression of CDK1. Restoration of CDK1 remarkably attenuated the inhibition of lycorine on the proliferation of NSCLC cells. SIGNIFICANCE: Our results uncovered the novel molecular mechanism of lycorine in suppressing the progression of NSCLC partially via regulating the miR-186/CDK1 axis.


Assuntos
Alcaloides de Amaryllidaceae/farmacologia , Proteína Quinase CDC2/metabolismo , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , MicroRNAs/metabolismo , Fenantridinas/farmacologia , Regiões 3' não Traduzidas , Apoptose/efeitos dos fármacos , Proteína Quinase CDC2/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , MicroRNAs/efeitos dos fármacos , MicroRNAs/genética , Regulação para Cima/efeitos dos fármacos
11.
Gene ; 709: 1-7, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31108165

RESUMO

Diabetes mellitus (DM) is a chronic, multifactorial metabolic disease whereby insulin deficiency or resistance results in hyperglycemia. A sustained high glucose environment results in inflammation and endothelial cell dysfunction. However, the underlying mechanisms are still not entirely clear. Circular RNAs (circRNAs) are recognized as functional non-coding RNAs involved in diverse biological processes, including DM. Previous studies have found that hsa_circ_0068087 is increased in DM patients. In order to identify whether hsa_circ_0068087 plays a role in high glucose (HG)-induced inflammation and endothelial cell dysfunction Human Umbilical Vein Endothelial Cell (HUVECs), quantitative reverse transcription PCR (qRTPCR), tube formation assay, enzyme-linked immunosorbent assay (ELISA) and bifluorescein reporter experiments were employed in this study. The results showed that the expression of hsa_circ_0068087 was upregulated in HUVECs following increases in glucose. Knockdown of hsa_circ_0068087 suppressed HG-induced HUVEC dysfunction and inflammation by suppression of the TLR4/NF-κB/NLRP3 inflammasome signaling pathway. Downregulation of miR-197 reversed hsa_circ_0068087 silence-induced HUVEC dysfunction and inflammation in the HG condition. It was found that TLR4 was the target of miR-197 and that overexpression of TLR4 ameliorated miR-197-induced HUVEC dysfunction and inhibited inflammation in the HG condition. Bifluorescein report experiments confirmed that miR-197 is a potential target of hsa_circ_0068087 and that TLR4 is a potential miR-197 target. Taken together, these results suggest that downregulation of hsa_circ_0068087 ameliorates TLR4/NF-κB/NLRP3 inflammasome-mediated inflammation and endothelial cell dysfunction in the high glucose condition by sponging miR-197.


Assuntos
Glucose/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Inflamassomos/fisiologia , Inflamação/genética , MicroRNAs/metabolismo , RNA/genética , Células Cultivadas , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiopatologia , Técnicas de Silenciamento de Genes , Humanos , Inflamassomos/efeitos dos fármacos , Inflamassomos/genética , Inflamassomos/metabolismo , Inflamação/metabolismo , MicroRNAs/efeitos dos fármacos , MicroRNAs/genética , RNA/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética
12.
eNeuro ; 6(2)2019.
Artigo em Inglês | MEDLINE | ID: mdl-31119189

RESUMO

Neuronal cholinergic circuits have been implicated in cognitive function and neurological disease, but the role of cholinergic signaling in other cellular populations within the brain has not been as fully defined. Here, we show that cholinergic signaling mechanisms are involved in mediating the function of the choroid plexus, the brain structure responsible for generating CSF and releasing various factors into the brain. The choroid plexus was found to express markers of endogenous cholinergic signaling, including multiple nicotinic acetylcholine receptor (nAChR) subtypes in a region-specific manner, and application of nicotine was found to induce cellular activation, as evidenced by calcium influx in primary tissue. During intravenous nicotine self-administration in male rats, nicotine increased expression of transthyretin, a protein selectively produced and released by the choroid plexus, and microRNA-204 (mir-204), a transcript found in high levels in the choroid plexus and CSF. Finally, human choroid plexus tissue from both sexes was found to exhibit similar nAChR, transthyretin and mir-204 expression profiles, supporting the translational relevance of the findings. Together, these studies demonstrate functionally active cholinergic signaling mechanisms in the choroid plexus, the resulting effects on transthyretin and mir-204 expression, and reveal the direct mechanism by which nicotine modulates function of this tissue.


Assuntos
Plexo Corióideo , MicroRNAs , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Pré-Albumina , Receptores Nicotínicos , Transdução de Sinais/efeitos dos fármacos , Animais , Plexo Corióideo/efeitos dos fármacos , Plexo Corióideo/metabolismo , Feminino , Humanos , Masculino , MicroRNAs/efeitos dos fármacos , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Pré-Albumina/efeitos dos fármacos , Pré-Albumina/metabolismo , Ratos , Ratos Wistar , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/metabolismo
13.
Methods Mol Biol ; 1974: 141-150, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31099000

RESUMO

Elevated levels of microRNAs in cancer cells are often associated with oncogenic effects and thus provide potential therapeutic targets. However, the lack of efficient delivery methods for synthetic miRNA inhibitors, antagomiR, or anti-miR oligonucleotides hindered clinical translation of such strategies. We recently developed an approach for targeted delivery of synthetic, 2'-O-methyl-modified antagomiR molecules to normal and malignant myeloid cells and B cells by tethering to the single-stranded, phosphorothioate oligodeoxynucleotides (PSO). The PSO-antagomiR are rapidly internalized through scavenger receptor-mediated endocytosis by human monocytes, dendritic cells, B cells, as well as myeloid leukemia and B-cell lymphoma cells, but not by T cells. Following internalization, the unformulated PSO-antagomiR potently reduces levels of target miRNA and modulates expression of downstream protein targets, both in vitro and in vivo. The simple design of PSO-antagomiR conjugates enable adaptation of this strategy for targeting oncogenic miRNAs in nonmalignant and malignant myeloid cells and B cells.


Assuntos
Antagomirs/farmacologia , Sistemas de Liberação de Medicamentos/métodos , Leucemia Mieloide/genética , Linfoma de Células B/genética , Animais , Linfócitos B , Humanos , Leucemia Mieloide/terapia , Linfoma de Células B/terapia , Camundongos , MicroRNAs/efeitos dos fármacos , MicroRNAs/genética , Células Mieloides/efeitos dos fármacos , Oligonucleotídeos Fosforotioatos/genética , Oligonucleotídeos Fosforotioatos/farmacologia , Linfócitos T/efeitos dos fármacos
14.
Acta Trop ; 195: 97-102, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31051116

RESUMO

Albendazole, as the main anti-echinococcal benzimidazole, has demonstrated safe and effective therapeutic outcomes in the treatment of echinococcosis. The emergence of resistance or reduced response to albendazole sulfoxide (ABZ_SOX) and other benzimidazoles have been demonstrated in several parasitic helminths of medical and veterinary importance. As the genetic makeup and miRNA profile of helminths affects their response to albendazole sulfoxide, the present study was conducted to investigate the expression of miRNAs in different developmental stages of Echinococcus granulosus exposed to albendazole sulfoxide in vitro. Different developmental stages of the helminth were obtained from in vitro cultured E. granulosus in monophasic and diphasic media. In both ABZ-SOX-treated and control parasites miRNAs were extracted from microcysts, intact protoscoleces and strobilated worms with one and three segments. Expression of two miRNAs, let-7 and miR-61 was evaluated using RT-qPCR for each stage. Results of the present study revealed significant differential expression of both let-7 and miR-61 at different drug concentrations. A significant difference of let-7 expression was observed between the strobilated and metacestode stages of E. granulosus exposed to ABZ-SOX. In the treated protoscoleces, let-7 expression was significantly reduced in the presence of ABZ-SOX at 1000 µg/ml concentration. In contrast higher expression levels were documented in the segmented worms. In the microcysts exposed to different drug concentrations a significant decline of miR-61 expression was demonstrated. Also, a significant increase in expression of miR-61 was observed in one proglottid worms as well as the protoscoleces. Under high drug concentration or long-term exposure of the protoscoleces to ABZ-SOX significantly higher miR-61 expression was observed compared to the controls. Our findings suggested that under in vitro benzimidazole exposure the expression of two E. granulosus miRNAs were significantly affected in the microcyst stage. This study presents the first evidence of the nature of benzimidazole effects on miRNA expression in platyhelminths.


Assuntos
Albendazol/uso terapêutico , Anti-Helmínticos/uso terapêutico , Equinococose/tratamento farmacológico , Echinococcus granulosus/efeitos dos fármacos , MicroRNAs/efeitos dos fármacos , Animais , Técnicas de Cultura de Células
15.
Genes (Basel) ; 10(4)2019 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-31022985

RESUMO

Nucleoside analog, cytarabine (ara-C) is the mainstay of acute myeloid leukemia (AML) chemotherapy. Cytarabine and other nucleoside analogs require activation to the triphosphate form (ara-CTP). Intracellular ara-CTP levels demonstrate significant inter-patient variation and have been related to therapeutic response in AML patients. Inter-patient variation in expression levels of drug transporters or enzymes involved in their activation or inactivation of cytarabine and other analogs is a prime mechanism contributing to development of drug resistance. Since microRNAs (miRNAs) are known to regulate gene-expression, the aim of this study was to identify miRNAs involved in regulation of messenger RNA expression levels of cytarabine pathway genes. We evaluated miRNA and gene-expression levels of cytarabine metabolic pathway genes in 8 AML cell lines and The Cancer Genome Atlas (TCGA) data base. Using correlation analysis and functional validation experiments, our data demonstrates that miR-34a-5p and miR-24-3p regulate DCK, an enzyme involved in activation of cytarabine and DCDT, an enzyme involved in metabolic inactivation of cytarabine expression, respectively. Further our results from gel shift assays confirmed binding of these mRNA-miRNA pairs. Our results show miRNA mediated regulation of gene expression levels of nucleoside metabolic pathway genes can impact interindividual variation in expression levels which in turn may influence treatment outcomes.


Assuntos
Antineoplásicos/farmacologia , Redes Reguladoras de Genes/efeitos dos fármacos , Leucemia Mieloide Aguda/tratamento farmacológico , MicroRNAs/genética , Nucleosídeos/análogos & derivados , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Citarabina/farmacologia , Citarabina/uso terapêutico , DCMP Desaminase/genética , Desoxicitidina Quinase/genética , Feminino , Perfilação da Expressão Gênica , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Células HL-60 , Humanos , Leucemia Mieloide Aguda/genética , Masculino , Redes e Vias Metabólicas , MicroRNAs/efeitos dos fármacos , Células THP-1
16.
Med Sci Monit ; 25: 3146-3153, 2019 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-31031401

RESUMO

BACKGROUND Estrogen levels regulate changes in osteoarthritis (OA) by inhibiting degradation of the extracellular matrix. Recent in vitro studies have also shown the role of microRNA-140-5p (miR-140-5p). This study aimed to investigate the role of estrogen deficiency, selective modulation of expression of the estrogen receptor (ER), and expression of miR-140-5p in cartilage and subchondral bone remodeling in an ovariectomized rat model of postmenopausal OA. MATERIAL AND METHODS Female Sprague-Dawley rats included two model groups, ovariectomized (OVX) rats and rats with destabilization of the medial meniscus (DMM) rats. Two months after surgery, estrogen levels were measured by the enzyme-linked immunosorbent assay (ELISA). Three-dimensional (3D) micro-computed tomography (micro-CT) was used to image the knee joints. Rats were treated with subcutaneous injection of estrogen (E2) or the selective estrogen receptor modulator (SERM), raloxifene (RAL), for one month. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect miR-140-5p in serum, and histology of the knee joint cartilage and bone was performed. RESULTS In the ovariectomized rat model of OA, estrogen therapy reduced the degree of cartilaginous degeneration, while treatment with raloxifene showed no significant effect. Expression levels of miR-140-5p in the OA model group were significantly lower than the control group. Micro-CT showed that in the model group, anterior cruciate ligament dislocation and subchondral bone density were significantly reduced. CONCLUSIONS In an ovariectomized rat model of postmenopausal OA, estrogen deficiency resulted in resorption of subchondral bone and degeneration of articular cartilage.


Assuntos
Estrogênios/administração & dosagem , Estrogênios/deficiência , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Animais , Ligamento Cruzado Anterior/cirurgia , Densidade Óssea , Remodelação Óssea , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Osso e Ossos/patologia , Cartilagem/metabolismo , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Modelos Animais de Doenças , Estrogênios/metabolismo , Matriz Extracelular/metabolismo , Feminino , Articulação do Joelho/cirurgia , MicroRNAs/sangue , MicroRNAs/efeitos dos fármacos , MicroRNAs/genética , Ovariectomia , Pós-Menopausa , Cloridrato de Raloxifeno/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores Estrogênicos/biossíntese
17.
Obes Facts ; 12(2): 211-225, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30999294

RESUMO

BACKGROUND: Increasing prevalence of obesity requires the investigation of respective comorbidities, including tumor diseases like colorectal, renal, post-menopausal breast, prostate cancer, and leukemia. To date, molecular mechanisms of the malignant transformation of these peripheral tissues induced by obesity remain unclear. Adipose tissue secretes factors with hormone-like functions, the adipokines, and is therefore categorized as an endocrine organ. Current research demonstrates the ability of adipose tissue to alter DNA methylation and gene expression in peripheral tissues, probably affecting microRNA (miR) expression. METHODS: Literature was analyzed for adipokine-regulated miRs. Many of these adipokine upregulated or downregulated miRs exert either oncogenic or anti-tumoral potential. RESULTS: The three selected and analyzed adipokines, adiponectin, leptin, and resistin, induce more strongly oncogenic miRs and simultaneously reduce anti-tumoral miRs than vice versa. This effect is not only true for the pure number of regulated miRs, it is also the case by consideration of the abundance of the respective miR expression based on actual data sets derived from next-generation sequencing. CONCLUSION: The link of obesity and cancer is analyzed under the aspect of adipokine-regulated miRs. At the same time the impact of miR abundance is considered as a regulatory variable. This context offers new strategies for tumor therapy and diagnostics.


Assuntos
Adipocinas/farmacologia , Genes Supressores de Tumor/efeitos dos fármacos , MicroRNAs/genética , Neoplasias/genética , Oncogenes/efeitos dos fármacos , Tecido Adiposo/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Leptina/metabolismo , Masculino , MicroRNAs/efeitos dos fármacos , Neoplasias/metabolismo , Obesidade/complicações , Obesidade/metabolismo , Oncogenes/genética
18.
Oncol Res ; 27(8): 889-899, 2019 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-30940289

RESUMO

The thorns of Gleditsia sinensis have been historically used in Chinese medicine and are considered one of the fundamental therapeutic herbs. Its anticancer effects are currently being explored. Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and still requires the development of new drugs with higher efficiency. By using a rat HCC model implanted with cancerous Walker-256 cells, the therapeutic effects of G. sinensis extract (GSE) were assessed, as well as its regulatory effects on miRNAs. GSE significantly restored liver morphology and dramatically induced cell apoptosis in HCC rats. In addition, miR-21/181b/183 was upregulated in the HCC liver, and the elevation of these miRNAs could be alleviated by both GSE and sorafenib. PTEN/TIMP3/PDCD4 downregulation was consistent with the targets of miR-21/181b/183 in the HCC liver, and the alteration of these target genes was restored by both GSE and sorafenib. TIMP3 effects on MMP-2/9 expression were also determined. Our present findings indicate the potential of GSE in HCC treatment, and expand the understanding of miRNA-related mechanisms in the anticancer effects of GSE.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Mamárias Animais/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/genética , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Gleditsia/química , Humanos , Neoplasias Hepáticas/patologia , Neoplasias Mamárias Animais/genética , Neoplasias Mamárias Animais/patologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , MicroRNAs/efeitos dos fármacos , PTEN Fosfo-Hidrolase/genética , Extratos Vegetais/farmacologia , Ratos , Inibidor Tecidual de Metaloproteinase-3/genética
19.
J Affect Disord ; 252: 32-38, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-30974330

RESUMO

BACKGROUND: MiRNAs are considered to be significant contributors to the pathogenesis of psychiatric diseases, but little is known about the potential roles of miRNAs in the treatment effect of panic disorder (PD). Therefore, we aimed to identify the miRNAs association with PD over the course of sertraline treatment. METHODS: Sixty-seven patients were collected for a 6-week period of sertraline treatment, and evaluated using HAMD-17, HAMA-14 and PDSS both at the baseline and 6 weeks later. Blood samples were collected before and after treatment, respectively. Ten pairs of samples were analyzed using miRNA array, and the differentially expressed miRNAs were further validated using RT-PCR in the whole sample. RESULTS: miR-451a, miR-144-5p, miR-25-3p and miR-660-5p were found to be significantly up-regulated, while miR-1 and miR-148-5p significantly down-regulated after sertraline treatment. The change of miR-25-3p before and after treatment (△miR-25-3p) was positively related to both the changes of PDSS3 scores (△PDSS3) (p = 0.017, 31.5% contribution) and △ PDSS7 (p = 0.016, 32.3% contribution). The △miR-660-5p was positively related to both the △HAMA5 (p = 0.03, 26% contribution) and △PDSS7 (p = 0.032). The △miR-148-5p was positively related to the △PDSS4 (p = 0.046, 21.5% contribution), but negatively related to the △HAMA13 (p = 0.005, 41.9% contribution). The △miR-144-5p was negatively related to the △HAMA9 (p = 0.032, 25.3% contribution). CONCLUSIONS: These findings might provide some evidences to the involvement of miRNA in the effect of anti-anxiety agents, which contributed to the better understanding the disease and developing new therapeutic genetic targets.


Assuntos
MicroRNAs/efeitos dos fármacos , Transtorno de Pânico/genética , Sertralina/farmacologia , Adolescente , Adulto , Antidepressivos/farmacologia , Antidepressivos/uso terapêutico , Regulação para Baixo/efeitos dos fármacos , Feminino , Humanos , Masculino , MicroRNAs/genética , Análise em Microsséries , Pessoa de Meia-Idade , Transtorno de Pânico/tratamento farmacológico , Sertralina/uso terapêutico , Regulação para Cima/efeitos dos fármacos , Adulto Jovem
20.
Exp Mol Pathol ; 108: 80-88, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30978333

RESUMO

BACKGROUND: Triptolide (TPL) is a potential anti-tumor natural compound. However, its role in nephroblastoma is poorly studied. Herein, we aimed to reveal the regulatory effects of TPL on human nephroblastoma cells (G-401 and WiT49) as well as the regulatory mechanism in G-401 cells. METHODS: Effects of TPL on cell viability, migration and apoptosis of G-401 and WiT49 cells were measured by CCK-8 assay, Boyden Chamber, and flow cytometry/Western blot analysis, respectively. Expression of miR-193b-3p in TPL-treated G-401 and WiT49 cells was determined by RT-qPCR. Then, whether miR-193b-3p was the downstream factor of TPL was studied. Alteration of KLF4 expression in TPL-treated cells and the relationship between miR-193b-3p and KLF4 were assessed by Western blot analysis and luciferase reporter assay. Effects of abnormally expressed KLF4 on G-401 cells were also assessed. Finally, the involvements of PI3K/AKT and ERK pathways were measured by Western blot analysis. RESULTS: TPL reduced cell viability and migration while promoted apoptosis in G-401 and WiT49 cells. miR-193b-3p level was up-regulated by TPL stimulation, and TPL might function through regulation of miR-193b-3p. KLF4 expression was down-regulated by TPL, and KLF4 was proven to be a target gene of miR-193b-3p. TPL mediated the repressive roles in growth of nephroblastoma cells via KLF4 knockdown. Finally, we found phosphorylation of PI3K, AKT and ERK was inhibited by TPL, possibly through miR-193b-3p-mediated regulation of KLF4. CONCLUSION: TPL showed a tumor suppressive role in nephroblastoma cells via miR-193b-3p-mediated down-regulation of KLF4, along with inhibition of the PI3K/AKT and ERK pathways.


Assuntos
Diterpenos/farmacologia , MicroRNAs/efeitos dos fármacos , Fenantrenos/farmacologia , Tumor de Wilms/genética , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Diterpenos/metabolismo , Compostos de Epóxi/metabolismo , Compostos de Epóxi/farmacologia , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Sistema de Sinalização das MAP Quinases/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Fenantrenos/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Tumor de Wilms/metabolismo
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