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1.
Anal Chem ; 93(4): 2152-2159, 2021 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-33406831

RESUMO

The macrophage migration inhibitory factor (MIF), a vital cytokine and biomarker, has been suggested to closely associate with the pathogenesis of liver cancer. However, a simple and effective approach for monitoring the change and distribution of cellular MIF is currently lacking and urgently needed, which could be helpful for a better understanding of its role in the progression of cancer. Herein, we report a novel activity-based probe, TPP2, which allows for direct labeling and imaging of endogenous MIF activity within live cells, clinical tissues, and in vivo in a mouse model of liver cancer. With this probe, we have intuitively observed the dynamic change of intracellular MIF activity by both flow cytometry and confocal imaging. We further found that TPP2 permits the identification and distinguishing of liver cancer in vitro and in vivo with high sensitivity and selectivity toward MIF. Our observations indicate that TPP2 could provide a promising new imaging approach for elucidating the MIF-related biological functions in liver cancer.


Assuntos
Fatores Inibidores da Migração de Macrófagos/metabolismo , Naftalenos/farmacologia , Animais , Linhagem Celular Tumoral , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Hepáticas/metabolismo , Masculino , Camundongos , Camundongos Nus , Naftalenos/química , Neoplasias Experimentais , Ligação Proteica , Conformação Proteica , Proteoma , Análise de Célula Única
2.
Int J Mol Sci ; 22(2)2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33430361

RESUMO

Anaplastic thyroid cancer (ATC) is an undifferentiated and advanced form of thyroid cancer, accompanied with a high ratio of epigenetic adjustment, which occurs more than genetic mutations. In this study, we aimed to evaluate the synergistic anticancer effect (in vitro and in vivo) of the new combination of N-hydroxy-7-(2-naphthylthio) heptanomide (HNHA) and sorafenib with radiation therapy in pre-clinical models of ATC. The ATC cell lines, YUMC-A1 and YUMC-A2, were isolated from the current patients who were treated with HNHA and sorafenib, either as monotherapy or combination therapy. Synergistic anticancer effect of the combination therapy on the intracellular signaling pathways and cell cycle was assessed via flow cytometry and immunoblot analysis. To examine tumor shrinkage activity in vivo, an ATC cell line-derived mouse xenograft model was used. Results showed that the combination therapy of HNHA and sorafenib with radiation promoted tumor suppression via caspase cleavage and cell cycle arrest in patient-derived ATC. In addition, the combination therapy of HNHA and sorafenib with radiation was more effective against ATC than therapy with HNHA or sorafenib with radiation. Thus, the combination of HNHA and sorafenib with radiation may be used as a novel curative approach for the treatment of ATC.


Assuntos
Proliferação de Células/efeitos dos fármacos , Ácidos Hidroxâmicos/farmacologia , Naftalenos/farmacologia , Sorafenibe/farmacologia , Carcinoma Anaplásico da Tireoide/tratamento farmacológico , Carcinoma Anaplásico da Tireoide/radioterapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos da radiação , Terapia Combinada , Sinergismo Farmacológico , Feminino , Xenoenxertos , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Radioterapia , Carcinoma Anaplásico da Tireoide/patologia
3.
J Ethnopharmacol ; 264: 113246, 2021 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-32781257

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Cyperus rotundus L. (Cyperaceae) is a widespread herbal in China and widely used in Traditional Chinese Medicine for multiple effects such as anti-arthritic, anti-genotoxic, anti-mutagenic, anti-bacterial effects, and analgesic. α-Cyperone is an active compound in Cyperus rotundus and has analgesic effects, but the exact molecular mechanisms require further investigations. MATERIALS AND METHODS: Tumor-derived DNA isolated from Lewis cell lines was transfected into microglia, and analyzed for stimulator of interferon genes (STING) effects. The downstream protein, such as interferon regulatory factor 3 (IRF3) and p65 nuclear factor-κB (NF-κB) were treated with STING siRNA and 5,6-dimethyllxanthenone-4-acetic acid (DMXAA) in microglia. The α-Cyperone effect on microglia was also investigated. RESULTS: Tumor-derived DNA activate microglia by upregulation of STING and downstream proteins. STING siRNA was reduced to its downstream expression and neuroinflammation inhibition was caused by tumor-derived DNA. However, DMXAA reversed the STING siRNA effect and increased neuroinflammation. α-Cyperone takes inhibitory effects on tumor-derived DNA that trigger microglia by STING pathway. CONCLUSIONS: α-Cyperone inhibition by tumor-derived DNA activated microglial to neuroinflammation in STING signaling pathway.


Assuntos
DNA de Neoplasias/antagonistas & inibidores , DNA de Neoplasias/genética , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , Microglia/efeitos dos fármacos , Naftalenos/farmacologia , Animais , Carcinoma Pulmonar de Lewis/tratamento farmacológico , Carcinoma Pulmonar de Lewis/genética , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Camundongos , Microglia/fisiologia , Naftalenos/uso terapêutico
4.
Gene ; 759: 145001, 2020 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-32738420

RESUMO

BACKROUND: CSCs having the common features of high telomerase activity and high migration and invasion capabilities play a vital role as the initiators of metastasis. Small molecule BIBR1532 has been shown to target cancer cells by inhibiting telomerase. Recent studies have suggested that telomerase activity is associated with epithelial mesenchymal transition (EMT). EMT program, which causes epithelial cells to acquire a mesenchymal morphology, is known to play a significant role in cancer metastasis. METHODS: The hypothesis of our study was that suppression of telomerase in breast cancer and cancer stem cells would interrupt EMT mechanism. Cytotoxicity of BIBR1532 was evaluated using WST-1 assay in all cell lines and the effects of BIBR1532 on apoptosis were investigated with Annexin V. Migration rate of the cells was examined by wound healing assay and sphere forming capacities were observed by hanging drop test. Finally, the expression of 84 EMT-related genes was analyzed by real-time qPCR. RESULTS: The IC50 values for the MDA-MB-231 and breast epithelial stem cells of BIBR1532 were analyzed as 18.04 and 38.71 µl at 72 h, respectively. Interestingly, apoptosis was only induced in stem cells. In hanging drop test, sphere areas were reduced in stem cells treated with BIBR1532. In wound healing assay, BIBR1532 decreased the migration rate of stem cells. Together with this, expression of EMT-related genes were regulated in stem cells towards a epithelial phenotype. CONCLUSION: Our obtained results indicated that telomerase inhibition affects the EMT mechanism. The targeted elimination of breast cancer stem cells by a telomerase inhibitor in cancer treatment may limit the mobility and stemness of cancer cells interrupting the EMT mechanism, thus may prevent metastasis.


Assuntos
Aminobenzoatos/farmacologia , Neoplasias da Mama/metabolismo , Inibidores Enzimáticos/farmacologia , Transição Epitelial-Mesenquimal , Naftalenos/farmacologia , Células-Tronco Neoplásicas/efeitos dos fármacos , Movimento Celular , Proliferação de Células , Feminino , Humanos , Células MCF-7 , Células-Tronco Neoplásicas/fisiologia , Telomerase/antagonistas & inibidores
5.
ACS Infect Dis ; 6(8): 2099-2109, 2020 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-32428392

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent for COVID-19, is a novel human betacoronavirus that is rapidly spreading worldwide. The outbreak currently includes over 3.7 million cases and 260,000 fatalities. As a betacoronavirus, SARS-CoV-2 encodes for a papain-like protease (PLpro) that is likely responsible for cleavage of the coronavirus (CoV) viral polypeptide. The PLpro is also responsible for suppression of host innate immune responses by virtue of its ability to reverse host ubiquitination and ISGylation events. Here, the biochemical activity of SARS-CoV-2 PLpro against ubiquitin (Ub) and interferon-stimulated gene product 15 (ISG15) substrates is evaluated, revealing that the protease has a marked reduction in its ability to process K48 linked Ub substrates compared to its counterpart in SARS-CoV. Additionally, its substrate activity more closely mirrors that of the PLpro from the Middle East respiratory syndrome coronavirus and prefers ISG15s from certain species including humans. Additionally, naphthalene based PLpro inhibitors are shown to be effective at halting SARS-CoV-2 PLpro activity as well as SARS-CoV-2 replication.


Assuntos
Betacoronavirus/enzimologia , Infecções por Coronavirus/virologia , Cisteína Endopeptidases/metabolismo , Citocinas/metabolismo , Enzimas Desubiquitinantes/antagonistas & inibidores , Pneumonia Viral/virologia , Ubiquitina/metabolismo , Ubiquitinas/metabolismo , Proteínas não Estruturais Virais/antagonistas & inibidores , Proteínas não Estruturais Virais/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Chlorocebus aethiops , Cisteína Endopeptidases/química , Citocinas/antagonistas & inibidores , Citocinas/química , Humanos , Naftalenos/farmacologia , Pandemias , Ligação Proteica , Conformação Proteica , Especificidade por Substrato , Ubiquitinas/antagonistas & inibidores , Ubiquitinas/química , Células Vero , Proteínas não Estruturais Virais/química , Replicação Viral/efeitos dos fármacos
6.
Sci Rep ; 10(1): 8535, 2020 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-32444682

RESUMO

Glaucoma is a group of optic neuropathies associated with aging and sensitivity to intraocular pressure (IOP). Early progression involves retinal ganglion cell (RGC) axon dysfunction that precedes frank degeneration. Previously we demonstrated that p38 MAPK inhibition abates axonal dysfunction and slows degeneration in the inducible microbead occlusion model of glaucoma in rat. Here, we assessed the neuroprotective effect of topical eye delivery of the p38 MAPK inhibitor BIRB 796 in three models of glaucoma (microbead occlusion in rat and squirrel monkey and the genetic DBA/2 J mouse model) with distinct durations of IOP elevation. While BIRB 796 did not influence IOP, treatment over four weeks in rats prevented degradation of anterograde axonal transport to the superior colliculus and degeneration in the optic nerve. Treatment over months in the chronic DBA/2 J model and in the squirrel monkey model reduced expression and activation of p38 downstream targets in the retina and brain but did not rescue RGC axon transport or degeneration, suggesting the efficacy of BIRB 796 in preventing associated degeneration of the RGC projection depends on the duration of the experimental model. These results emphasize the importance of evaluating potential therapeutic compounds for neuroprotection in multiple models using elongated treatment paradigms for an accurate assessment of efficacy.


Assuntos
Glaucoma/tratamento farmacológico , Naftalenos/farmacologia , Fármacos Neuroprotetores/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Pirazóis/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Transporte Axonal/efeitos dos fármacos , Modelos Animais de Doenças , Pressão Intraocular/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos DBA , Nervo Óptico/efeitos dos fármacos , Nervo Óptico/metabolismo , Ratos , Células Ganglionares da Retina/efeitos dos fármacos , Células Ganglionares da Retina/metabolismo , Saimiri
7.
Proc Natl Acad Sci U S A ; 117(18): 9991-10002, 2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32312805

RESUMO

The initial response to an addictive substance can facilitate repeated use: That is, individuals experiencing more positive effects are more likely to use that drug again. Increasing evidence suggests that psychoactive cannabinoid use in adolescence enhances the behavioral effects of cocaine. However, despite the behavioral data, there is no neurobiological evidence demonstrating that cannabinoids can also alter the brain's initial molecular and epigenetic response to cocaine. Here, we utilized a multiomics approach (epigenomics, transcriptomics, proteomics, and phosphoproteomics) to characterize how the rat brain responds to its first encounter with cocaine, with or without preexposure to the synthetic cannabinoid WIN 55,212-2 (WIN). We find that in adolescent (but not in adult) rats, preexposure to WIN results in cross-sensitization to cocaine, which correlates with histone hyperacetylation and decreased levels of HDAC6 in the prefrontal cortex (PFC). In the PFC, we also find that WIN preexposure blunts the typical mRNA response to cocaine and instead results in alternative splicing and chromatin accessibility events, involving genes such as Npas2 Moreover, preexposure to WIN enhances the effects of cocaine on protein phosphorylation, including ERK/MAPK-targets like gephyrin, and modulates the synaptic AMPAR/GluR composition both in the PFC and the nucleus accumbens (NAcc). PFC-NAcc gene network topological analyses, following cocaine exposure, reveal distinct top nodes in the WIN preexposed group, which include PACAP/ADCYAP1. These preclinical data demonstrate that adolescent cannabinoid exposure reprograms the initial behavioral, molecular, and epigenetic response to cocaine.


Assuntos
Comportamento Aditivo/genética , Comportamento Animal/efeitos dos fármacos , Canabinoides/efeitos adversos , Cocaína/efeitos adversos , Adolescente , Animais , Comportamento Aditivo/induzido quimicamente , Comportamento Aditivo/patologia , Benzoxazinas/efeitos adversos , Benzoxazinas/farmacologia , Canabinoides/farmacologia , Peptídeos e Proteínas de Sinalização do Ritmo Circadiano/genética , Cocaína/farmacologia , Epigênese Genética/efeitos dos fármacos , Epigênese Genética/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Desacetilase 6 de Histona/genética , Humanos , Proteínas de Membrana/farmacologia , Morfolinas/efeitos adversos , Morfolinas/farmacologia , Naftalenos/efeitos adversos , Naftalenos/farmacologia , Fosfoproteínas/efeitos dos fármacos , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/genética , Córtex Pré-Frontal/efeitos dos fármacos , Proteoma/efeitos dos fármacos , Ratos , Transcriptoma/efeitos dos fármacos
8.
J Med Chem ; 63(12): 6513-6522, 2020 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-32223238

RESUMO

Orteronel (TAK-700) is a substituted imidazole that was developed for the treatment of castration-resistant prostate cancer but was dropped in phase III clinical trials. Both enantiomers of this inhibitor of cytochrome P450 (P450) 17A1 show some selectivity in differentially blocking the 17α-hydroxylation and lyase activities of the enzyme. Although both enantiomers of this compound have sub-micromolar IC50 values and bind to the enzyme with a type II spectral change (indicative of nitrogen-iron bonding) and reported Kd values of 56 and 40 nM (R and S, respectively), the rates of binding to P450 17A1 were relatively slow. We considered the possibility that the drug is a slow, tight-binding inhibitor. Analysis of the kinetics of binding revealed rapid formation of an initial complex, presumably in the substrate binding site, followed by a slower change to the spectrum of a final iron complex. Similar kinetics were observed in the interaction of another inhibitor, the triazole (S)-seviteronel (VT-464), with P450 17A1. Kinetic tests and modeling indicate that the further change to the iron-complexed form of the orteronel- or seviteronel-P450 complex is not a prerequisite for enzyme inhibition. Accordingly, the inclusion of heme-binding heterocyclic nitrogen moieties in P450 17A1 inhibitors may not be necessary to achieve inhibition but may nevertheless augment the process.


Assuntos
Inibidores Enzimáticos/farmacologia , Imidazóis/farmacologia , Naftalenos/farmacologia , Esteroide 17-alfa-Hidroxilase/antagonistas & inibidores , Triazóis/farmacologia , Antagonistas de Receptores de Andrógenos/farmacologia , Sítios de Ligação , Humanos , Hidroxilação , Cinética , Oxirredução , Conformação Proteica , Esteroide 17-alfa-Hidroxilase/metabolismo
9.
J Bone Miner Metab ; 38(5): 687-694, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32274572

RESUMO

INTRODUCTION: Primary hyperparathyroidism (PHPT) is caused by parathyroid adenoma, primary parathyroid hyperplasia, or parathyroid carcinoma. For some patients with PHPT controlling serum calcium levels is critical. MATERIALS AND METHODS: We conducted an open-label, single-arm, 52-week, phase III study in Japanese patients with hypercalcemia due to PHPT to demonstrate efficacy and safety of evocalcet, a new calcimimetic. Patients with intractable PHPT (n = 13), postsurgical recurrence (n = 2), and parathyroid carcinoma (n = 3) were enrolled. Evocalcet administration started at a dose of 2 mg once or twice daily and was titrated to achieve the target serum corrected calcium (cCa) concentration (≤ 10.3 mg/dL) for two consecutive weeks (maximal dose 24 mg/day). RESULTS: Fourteen patients achieved the target (77.8%; 95% confidence interval [CI] 52.4-93.6). The lower limit of 95% CI exceeded the predetermined reference limit (11%), and thus, efficacy was confirmed. Of 18 patients, 12 (66.7%; 95% CI 41.0-86.7) showed decreased serum cCa of ≥ 1.0 mg/dL from the baseline for two consecutive weeks during the titration phase. Sixteen patients entered the maintenance phase, and 15 patients completed the study. Treatment-emergent adverse events (TEAEs) were recorded in 18/18 patients (100%) and drug-related TEAEs in 8/18 (44.4%). The most commonly observed drug-related TEAE was nausea (2/18 patients). No unexpected drug-related TEAEs were observed. All drug-related TEAEs were mild in severity. No patient discontinued the study because of drug-related TEAEs. CONCLUSION: Evocalcet demonstrated long-term effectiveness in reducing serum cCa concentrations and safety without any unexpected drug-related TEAEs in PHPT patients.


Assuntos
Hiperparatireoidismo Primário/tratamento farmacológico , Naftalenos/uso terapêutico , Pirrolidinas/uso terapêutico , Cálcio/sangue , Creatinina/sangue , Relação Dose-Resposta a Droga , Feminino , Humanos , Hiperparatireoidismo Primário/sangue , Masculino , Pessoa de Meia-Idade , Naftalenos/administração & dosagem , Naftalenos/efeitos adversos , Naftalenos/farmacologia , Hormônio Paratireóideo/sangue , Fosfatos/sangue , Pirrolidinas/administração & dosagem , Pirrolidinas/efeitos adversos , Pirrolidinas/farmacologia
10.
Nat Commun ; 11(1): 1830, 2020 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-32286350

RESUMO

A synthetic biology method based on heterologous biosynthesis coupled with genome mining is a promising approach for increasing the opportunities to rationally access natural product with novel structures and biological activities through total biosynthesis and combinatorial biosynthesis. Here, we demonstrate the advantage of the synthetic biology method to explore biological activity-related chemical space through the comprehensive heterologous biosynthesis of fungal decalin-containing diterpenoid pyrones (DDPs). Genome mining reveals putative DDP biosynthetic gene clusters distributed in five fungal genera. In addition, we design extended DDP pathways by combinatorial biosynthesis. In total, ten DDP pathways, including five native pathways, four extended pathways and one shunt pathway, are heterologously reconstituted in a genetically tractable heterologous host, Aspergillus oryzae, resulting in the production of 22 DDPs, including 15 new analogues. We also demonstrate the advantage of expanding the diversity of DDPs to probe various bioactive molecules through a wide range of biological evaluations.


Assuntos
Diterpenos/farmacologia , Fungos/química , Naftalenos/farmacologia , Pironas/farmacologia , Biologia Sintética , Peptídeos beta-Amiloides/metabolismo , Animais , Fármacos Anti-HIV/farmacologia , Aspergillus/química , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Proliferação de Células/efeitos dos fármacos , Diterpenos/química , Drosophila/efeitos dos fármacos , Fungos/genética , Genoma Fúngico , HIV-1/efeitos dos fármacos , Humanos , Células MCF-7 , Naftalenos/química , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Agregados Proteicos , Pironas/química , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/metabolismo , Esferoides Celulares/patologia , Estereoisomerismo
11.
Biochem Biophys Res Commun ; 525(3): 639-645, 2020 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-32122652

RESUMO

Hypoxia training (HT) can reduce body weight and improve fatty liver. However, the mechanism is not clear. A previous study indicated that HT-induced weight loss might be associated with the endocannabinoid system (ECS), which has also been reported recently to be involved in the persistent lipid mediators after weight loss. The present study investigated the effects of HT, a new prospective weight-loss method, on nutritionally obese mice and demonstrated that HT significantly reduced body weight, fat mass, transcriptional expression of liver endocannabinoid receptor 1 (CB1), biosynthetic enzyme diacylglycerol lipase α (DAGLα) and improved the transcriptional expression of degrading enzyme monoacylglycerol lipase (MAGL). Liver endocannabinoids 2-arachidonoylglycerol (2-AG) but not anandamide (AEA) was evidently decreased in response to HT. Simultaneously, HT significantly reduced liver index, serum alanine aminotransferase (ALT) and liver fat contents. Western blot showed decreased expression of liver CB1, sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor γ (PPARγ) and increased expression of adipose triglyceride lipase (ATGL) and carnitine palmitoyltransferase-1 (CPT-1) levels after HT. However, intraperitoneal injection of CB1 receptor agonist WIN55212-2 offset the benefits by which HT reduced hepatic fat synthesis, with significant increased protein expression of SREBP-1 and PPARγ. Taken together, these findings reported the alleviation of obesity and hepatic steatosis through HT and provided a putative molecular mechanism by inhibiting the CB1-mediated fat synthesis.


Assuntos
Regulação para Baixo , Fígado Gorduroso/genética , Fígado Gorduroso/patologia , Hipóxia/patologia , Receptor CB1 de Canabinoide/metabolismo , Animais , Benzoxazinas/farmacologia , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Endocanabinoides/metabolismo , Hipóxia/genética , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Obesos , Morfolinas/farmacologia , Naftalenos/farmacologia , Fenótipo , Receptor CB1 de Canabinoide/agonistas
12.
Int J Mol Sci ; 21(6)2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32183038

RESUMO

A focused library of newly designed monomeric and dimeric naphthalene diimides (NDIs) was analyzed in its ability to recognize specific G-quadruplex (G4) structures discriminating duplex DNA. The best G4 ligands-according to an affinity chromatography-based screening method named G4-CPG-were tested on human cancer and healthy cells, inducing DNA damage at telomeres, and in parallel, showing selective antiproliferative activity on HeLa cancer cells with IC50 values in the low nanomolar range. CD and fluorescence spectroscopy studies allowed detailed investigation of the interaction in solution with different G4 and duplex DNA models of the most promising NDI of the series, as determined by combining the biophysical and biological assays' data.


Assuntos
Antineoplásicos/química , Quadruplex G/efeitos dos fármacos , Iminas/química , Naftalenos/química , Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Dano ao DNA , Células HeLa , Humanos , Iminas/farmacologia , Ligantes , Naftalenos/farmacologia , Telômero/efeitos dos fármacos
13.
PLoS One ; 15(3): e0230753, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32218595

RESUMO

Rabbits (Oryctolagus cuniculi) are very popular as pets. However, problems of otitits caused by Psoroptes cuniculi are one of the main reasons to visit the veterinarian. Isoxazolines are an alternative treatment to treat this mite, and therefore, an evaluation of the effectiveness of oral afoxalaner with milbemycin oxime in rabbits infected with P. cuniculi was carried out. Nineteen rabbits, of New Zealand breed, with otitis due to an infection with P. cuniculi, were treated, whereas six rabbits were left untreated and formed the control group. The ear canals of each individual were examined, through the collection of otic exudate samples with cotton swabs. These were visualized under the microscope to identify the ectoparasite. Each animal was treated with a single oral dose of 2.50 mg / kg of afoxolaner, and 0.50 mg / kg of milbemycin oxime. Clinical signs and lesions associated with the infection, such as the presence of detritus, cerumen and / or scabs, and erythema, were evaluated. After receiving the treatment, all the lesions were classified as: mild, moderate and intense, with a visual analog scale. A week after providing medication, there was a decrease in the lesions of the group treated with Nexgard Spectra®, without further topical or systemic treatment. The decrease was gradual in the treated group and no recurrence was detected of P. cuniculi infection in both ears. Thus, the administration of a single oral dose of afoxolaner with milbemycin oxime was effective for the treatment of P. cuniculi infection in rabbits.


Assuntos
Isoxazóis/farmacologia , Macrolídeos/farmacologia , Infestações por Ácaros/tratamento farmacológico , Naftalenos/farmacologia , Psoroptidae/fisiologia , Animais , Relação Dose-Resposta a Droga , Interações Medicamentosas , Isoxazóis/uso terapêutico , Macrolídeos/uso terapêutico , Naftalenos/uso terapêutico , Coelhos
14.
Invest Ophthalmol Vis Sci ; 61(3): 3, 2020 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-32150246

RESUMO

Purpose: In the mammalian retina, cannabinoid type 1 receptors (CB1Rs) are well-positioned to alter inhibitory synaptic function from amacrine cells and, thus, might influence visual signal processing in the inner retina. However, it is not known if CB1R modulates amacrine cells feedback inhibition at retinal bipolar cell (BC) terminals. Methods: Using whole-cell voltage-clamp recordings, we examined the pharmacological effect of CB1R activation and inhibition on spontaneous inhibitory postsynaptic currents (sIPSCs) and glutamate-evoked IPSCs (gIPSCs) from identified OFF BCs in light-adapted rat retinal slices. Results: Activation of CB1R with WIN55212-2 selectively increased the frequency of GABAergic, but not glycinergic sIPSC in types 2, 3a, and 3b OFF BCs, and had no effect on inhibitory activity in type 4 OFF BCs. The increase in GABAergic activity was eliminated in axotomized BCs and can be suppressed by blocking CB1R with AM251 or GABAA and GABAρ receptors with SR-95531 and TPMPA, respectively. In all OFF BC types tested, a brief application of glutamate to the outer plexiform layer elicited gIPSCs comprising GABAergic and glycinergic components that were unaffected by CB1R activation. However, blocking CB1R selectively increased GABAergic gIPSCs, supporting a role for endocannabinoid signaling in the regulation of glutamate-evoked GABAergic inhibitory feedback to OFF BCs. Conclusions: CB1R activation shape types 2, 3a, and 3b OFF BC responses by selectively regulate GABAergic feedback inhibition at their axon terminals, thus cannabinoid signaling might play an important role in the fine-tuning of visual signal processing in the mammalian inner retina.


Assuntos
Receptor CB1 de Canabinoide/fisiologia , Células Bipolares da Retina/fisiologia , Células Amácrinas/metabolismo , Células Amácrinas/fisiologia , Animais , Benzoxazinas/farmacologia , Polaridade Celular/efeitos dos fármacos , Polaridade Celular/fisiologia , Endocanabinoides/metabolismo , Retroalimentação Fisiológica/efeitos dos fármacos , Retroalimentação Fisiológica/fisiologia , Feminino , Antagonistas de Receptores de GABA-A/farmacologia , Ácido Glutâmico/farmacologia , Potenciais Pós-Sinápticos Inibidores/efeitos dos fármacos , Potenciais Pós-Sinápticos Inibidores/fisiologia , Masculino , Morfolinas/farmacologia , Naftalenos/farmacologia , Técnicas de Patch-Clamp/métodos , Ácidos Fosfínicos/farmacologia , Piperidinas/farmacologia , Pirazóis/farmacologia , Piridinas/farmacologia , Ratos Sprague-Dawley , Receptor CB1 de Canabinoide/antagonistas & inibidores , Receptor CB1 de Canabinoide/efeitos dos fármacos , Retina , Células Bipolares da Retina/efeitos dos fármacos , Transdução de Sinais/fisiologia
15.
Int J Mol Sci ; 21(4)2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32098220

RESUMO

Histone deacetylases have been a target of therapy for organ fibrosis. Here, we report the protective effect of CG200745 (CG), a novel histone deacetylase inhibitor, on tubulointerstitial fibrosis in Col4a3-/- mice, a murine model of Alport syndrome. Morphological analyses revealed CG treatment markedly alleviated kidney fibrosis in Col4a3-/- mice at the age of 7 weeks. CG prevented the activation of transforming growth factor ß (TGFß) and its downstream SMAD signaling in the kidney of Col4a3-/- mice. As critical upstream regulators of TGFß signaling, immunoblotting of whole kidney lysate of Col4a3-/- mice reveled that intra-renal renin-angiotensin system (RAS) was activated with concurrent upregulation of inflammation and apoptosis, which were effectively suppressed by CG treatment. CG suppressed both activation of RAS and up-regulation of TGFß signals in angiotensin II-stimulated HK-2 cells, a human kidney proximal tubular epithelial cell line. CG inhibited activation of TGFß-driven signals and fibrosis in NRK-49F cells, a rat kidney fibroblast cell line, under angiotensin II-rich conditions. Collectively, CG was found to be effective both in proximal tubular epithelial cells by inhibiting local RAS and TGFß signaling activation, as well as in fibroblasts by blocking their transition to myofibroblasts, attenuating renal fibrosis in a murine model of Alport syndrome.


Assuntos
Inibidores de Histona Desacetilases/farmacologia , Ácidos Hidroxâmicos/farmacologia , Túbulos Renais Proximais/metabolismo , Naftalenos/farmacologia , Nefrite Hereditária , Transdução de Sinais , Animais , Autoantígenos/metabolismo , Linhagem Celular , Colágeno Tipo IV/deficiência , Colágeno Tipo IV/metabolismo , Modelos Animais de Doenças , Fibrose , Humanos , Túbulos Renais Proximais/patologia , Masculino , Camundongos , Camundongos Knockout , Nefrite Hereditária/tratamento farmacológico , Nefrite Hereditária/genética , Nefrite Hereditária/metabolismo , Nefrite Hereditária/patologia , Ratos , Fator de Crescimento Transformador beta/metabolismo
16.
Int J Mol Sci ; 21(4)2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-32074976

RESUMO

In the process of neonatal encephalopathy, oxidative stress and neuroinflammation have a prominent role after perinatal asphyxia. With the exception of therapeutic hypothermia, no therapeutic interventions are available in the clinical setting to target either the oxidative stress or inflammation, despite the high prevalence of neurological sequelae of this devastating condition. The endocannabinoid system (ECS), recently recognized as a widespread neuromodulatory system, plays an important role in the development of the central nervous system (CNS). This study aims to evaluate the potential effect of the cannabinoid (CB) agonist WIN 55,212-2 (WIN) on reactive oxygen species (ROS) and early inflammatory cytokine production after hypoxia-ischemia (HI) in fetal lambs. Hypoxic-ischemic animals were subjected to 60 min of HI by partial occlusion of the umbilical cord. A group of lambs received a single dose of 0.01 µg/kg WIN, whereas non-asphyctic animals served as controls. WIN reduced the widespread and notorious increase in inflammatory markers tumor necrosis factor (TNF)-α and interleukin (IL)-1ß and IL-6 induced by HI, a modulatory effect not observed for oxidative stress. Our study suggests that treatment with a low dose of WIN can alter the profile of pro-inflammatory cytokines 3 h after HI.


Assuntos
Canabinoides/farmacologia , Citocinas/metabolismo , Hipóxia-Isquemia Encefálica/patologia , Estresse Oxidativo/efeitos dos fármacos , Animais , Benzoxazinas/farmacologia , Córtex Cerebral/metabolismo , Córtex Cerebral/fisiologia , Modelos Animais de Doenças , Feminino , Feto/metabolismo , Hipóxia-Isquemia Encefálica/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Morfolinas/farmacologia , Naftalenos/farmacologia , Gravidez , Espécies Reativas de Oxigênio/química , Espécies Reativas de Oxigênio/metabolismo , Receptores de Canabinoides/química , Receptores de Canabinoides/metabolismo , Ovinos , Fator de Necrose Tumoral alfa/metabolismo
17.
Molecules ; 25(4)2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32085460

RESUMO

: Multiple drug resistant fungi pose a serious threat to human health, therefore the development of completely new antimycotics is of paramount importance. The in vitro antifungal activity of the original, 1-amino-5-isocyanonaphthalenes (ICANs) was evaluated against reference strains of clinically important Candida species. Structure-activity studies revealed that the naphthalene core and the isocyano- together with the amino moieties are all necessary to exert antifungal activity. 1,1-N-dimethylamino-5-isocyanonaphthalene (DIMICAN), the most promising candidate, was tested further in vitro against clinical isolates of Candida species, yielding a minimum inhibitory concentration (MIC) of 0.04-1.25 µg/mL. DIMICAN was found to be effective against intrinsically fluconazole resistant Candida krusei isolates, too. In vivo experiments were performed in a severly neutropenic murine model inoculated with a clinical strain of Candida albicans. Daily administration of 5 mg/kg DIMICAN intraperitoneally resulted in 80% survival even at day 13, whereas 100% of the control group died within six days. Based on these results, ICANs may become an effective clinical lead compound family against fungal pathogens.


Assuntos
Antifúngicos/farmacologia , Naftalenos/farmacologia , Animais , Antifúngicos/química , Antifúngicos/uso terapêutico , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Candidíase/tratamento farmacológico , Candidíase/microbiologia , Linhagem Celular , Modelos Animais de Doenças , Feminino , Humanos , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Naftalenos/química , Naftalenos/uso terapêutico , Relação Estrutura-Atividade
18.
Molecules ; 25(3)2020 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-32033198

RESUMO

G-quadruplex specific targeting molecules, also termed as G4 ligands, are attracting increasing attention for their ability to recognize and stabilize G-quadruplex and high potentiality for biological regulation. However, G4 ligands recognizing G-quadruplex were generally investigated within a dilute condition, which might be interfered with under a cellular crowding environment. Here, we designed and synthesized several new cyclic naphthalene diimide (cNDI) derivatives, and investigated their interaction with G-quadruplex under molecular crowding condition (40% v/v polyethylene glycol (PEG)200) to mimic the cellular condition. The results indicated that, under molecular crowding conditions, cNDI derivatives were still able to recognize and stabilize G-quadruplex structures based on circular dichroism measurement. The binding affinities were slightly decreased but still comparatively high upon determination by isothermal titration calorimetry and UV-vis absorbance spectroscopy. More interestingly, cNDI derivatives were observed with preference to induce a telomere sequence to form a hybrid G-quadruplex under cation-deficient molecular crowding conditions.


Assuntos
DNA/química , DNA/metabolismo , Imidas/síntese química , Imidas/farmacologia , Naftalenos/síntese química , Naftalenos/farmacologia , Calorimetria , Dicroísmo Circular , Quadruplex G , Humanos , Imidas/química , Estrutura Molecular , Naftalenos/química , Polietilenoglicóis/química , Potássio , Proteínas Proto-Oncogênicas c-myc/química , Proteínas Proto-Oncogênicas c-myc/metabolismo , Telômero/química , Telômero/metabolismo
19.
Exp Cell Res ; 389(1): 111881, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32006556

RESUMO

Human adipose tissue includes large quantities of mesenchymal stromal cells (atMSCs), which represent an abundant cell source for therapeutic applications in the field of regenerative medicine. Adipose tissue secrets various soluble factors including endocannabinoids, and atMSCs express the cannabinoid receptors CB1 and CB2. This indicates that adipose tissue possesses an endocannabinoid system (ECS). The ECS is also ascribed great significance for wound repair, e.g. by modulating inflammation. However, the exact effects of CB1/CB2 activation in human atMSCs have not been investigated, yet. In the present study, we stimulated human atMSCs with increasing concentrations (1-30 µM) of the unspecific cannabinoid receptor ligand WIN55,212-2 and the specific CB2 agonist JWH-133, either alone or co-applied with the receptor antagonist Rimonabant (CB1) or AM 630 (CB2). We investigated the effects on metabolic activity, cell number, differentiation and cytokine release, which are important processes during tissue regeneration. WIN decreased metabolic activity and cell number, which was reversed by Rimonabant. This suggests a CB1 dependent mechanism, whereas the number of atMSCs was increased after CB2 ligation. WIN and JWH increased the release of VEGF, TGF-ß1 and HGF. Adipogenesis was enhanced by WIN, which could be reversed by blocking CB1. There was no effect on osteogenesis, and only WIN increased chondrogenic differentiation. Our results indicate that definite activation of the cannabinoid receptors exerted different effects in atMSCs, which could be of specific value in cell-based therapy for wound regeneration.


Assuntos
Tecido Adiposo/citologia , Autorrenovação Celular , Células-Tronco Mesenquimais/fisiologia , Receptor CB1 de Canabinoide/fisiologia , Receptor CB2 de Canabinoide/fisiologia , Regeneração , Benzoxazinas/farmacologia , Canabinoides/farmacologia , Diferenciação Celular/efeitos dos fármacos , Autorrenovação Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Endocanabinoides/agonistas , Endocanabinoides/antagonistas & inibidores , Endocanabinoides/farmacologia , Humanos , Indóis/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Morfolinas/farmacologia , Naftalenos/farmacologia , Cultura Primária de Células , Receptor CB1 de Canabinoide/agonistas , Receptor CB1 de Canabinoide/antagonistas & inibidores , Receptor CB2 de Canabinoide/agonistas , Receptor CB2 de Canabinoide/antagonistas & inibidores , Regeneração/efeitos dos fármacos , Regeneração/fisiologia , Rimonabanto/farmacologia
20.
Pharmacol Rep ; 72(1): 115-125, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32016832

RESUMO

BACKGROUND: Although several studies had addressed the anti-inflammatory effects of derivatives of 4H-chromene and chromeno[2,3-b]pyridine in the different types of cells, whether these derivatives would exert beneficial anti-fibrotic effects during corneal fibrotic scar formation was unclear. METHODS: We examined the cyclooxygenase-2 (COX-2) expression of 2,4-diamino-5-(1-hydroxynaphthalen-2-yl)-5H-chromeno[2,3-b]pyridine-3-carbonitrile (N1) in the human corneal fibroblasts (HCFs) under the treatment TGF-ß1. Signaling pathways underlying the mechanism of the N1 effect on the HCFs were determined. RESULTS: Application of N1 significantly decreased COX-2 expression after 2 h and 4 h in the HCFs stimulated with TGF-ß1. Notably, reduced production of extracellular matrix proteins under N1 treatment was found, including fibronectin, collagen I, and matrix metallopeptidase 9. Immunoblot analysis showed that treatment with N1 significantly attenuated phosphorylation of both STAT3 and Smad 2 in the TGF-ß1-stimulated HCFs. Upregulated mRNA of Smad2 and downregulated mRNA of Smad3 were observed using the quantitative real-time polymerase chain reaction. In addition, N1 induced significant increases in HO-1 and Nrf2 expression, but inhibited phosphorylation of NF-κB in the HCFs treated with TGF-ß1. CONCLUSIONS: Our findings show for the first time that N1 exerts anti-fibrotic and anti-inflammatory effects through suppression of COX-2, Smad2, STAT3, iNOS and NF-κB expressions as well as upregulation of Nrf2 and HO-1 expressions, which suggests they are potential therapeutic targets in the treatment of corneal fibrosis.


Assuntos
Anti-Inflamatórios/farmacologia , Córnea/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Naftalenos/farmacologia , Células Cultivadas , Córnea/citologia , Regulação para Baixo/efeitos dos fármacos , Fibroblastos/citologia , Fibrose/patologia , Humanos , Inflamação/tratamento farmacológico , Inflamação/patologia , Transdução de Sinais/efeitos dos fármacos , Fator de Crescimento Transformador beta1/administração & dosagem , Regulação para Cima/efeitos dos fármacos
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