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1.
J Agric Food Chem ; 67(44): 12219-12227, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31613626

RESUMO

Quantification, using an accurate analytical approach, of capsinoids and capsaicinoids was performed on three chili pepper (Capsicum spp.) genotypes: "Chiltepín", "Tampiqueño 74", and "Bhut Jolokia" at various stages of fruit development. The accumulation of capsinoids, in all these peppers started between 10 to 20 days post-anthesis (dpa), increased and reached the highest capsinoid amount at 40 dpa, and then decreased until 60 dpa. Conversely, capsaicinoids could already be determined at 10 dpa in "Bhut Jolokia" and their accumulation pattern was different from that of the capsinoids in this genotype. The capsiate/dihydrocapsiate ratio presented a higher variation between genotypes and developmental stages than the capsaicin/dihydrocapsaicin ratio. Capsinoid ratios (4-24%) and Pun1/pAMT genotyping were determined. These results provide information on the progress of the accumulation of capsinoids in the aforementioned pungent and superhot cultivars and could support future breeding studies toward the understanding of the factors affecting their accumulation.


Assuntos
Capsaicina/análogos & derivados , Capsaicina/metabolismo , Capsicum/genética , Capsicum/metabolismo , Aromatizantes/metabolismo , Frutas/crescimento & desenvolvimento , Proteínas de Plantas/genética , Alelos , Sequência de Aminoácidos , Capsaicina/análise , Capsicum/química , Capsicum/crescimento & desenvolvimento , Aromatizantes/análise , Frutas/química , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Genótipo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alinhamento de Sequência
2.
Br J Anaesth ; 123(4): 439-449, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31383364

RESUMO

BACKGROUND: Nerve growth factor (NGF) has been implicated in hyperalgesia by sensitising nociceptors. A role for NGF in modulating myocardial injury through ischaemic nociceptive signalling is plausible. We examined whether inhibition of spinal NGF attenuates myocardial ischaemia-reperfusion injury and explored the underlying mechanisms. METHODS: In adult rats, lentivirus-mediated short-hairpin RNA targeted at reducing NGF gene expression (NGF-shRNA) or a transient receptor potential vanilloid 1 (TRPV1) antagonist (capsazepine) was injected intrathecally before myocardial ischaemia-reperfusion. Infarct size (expressed as the ratio of area at risk) and risk of arrhythmias were quantified. Whole-cell clamp patch electrophysiology was used to record capsaicin currents in primary dorsal root ganglion neurones. The co-expression of substance P (SP) and calcitonin gene-related peptide (CGRP), plus activation of TRPV1, protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) were also quantified. RESULTS: NGF levels increased by 2.95 (0.34)-fold in dorsal root ganglion and 2.12 (0.27)-fold in spinal cord after myocardial ischaemia-reperfusion injury. Intrathecal injection of NGF-shRNA reduced infarct area at risk from 0.58 (0.02) to 0.37 (0.02) (P<0.01) and reduced arrhythmia score from 3.67 (0.33) to 1.67 (0.33) (P<0.01). Intrathecal capsazepine was similarly cardioprotective. NGF-shRNA suppressed expression of SP/CGRP and activation of Akt/ERK and TRPV1 in spinal cord. NGF increased capsaicin current amplitude from 144 (42) to 840 (132) pA (P<0.05), which was blocked by the TRPV1 antagonist 5'-iodoresiniferatoxin. Exogenous NGF enhanced capsaicin-induced Akt/ERK and TRPV1 activation in PC12 neuroendocrine tumour cells in culture. CONCLUSIONS: Spinal NGF contributes to myocardial ischaemia-reperfusion injury by mediating nociceptive signal transmission.


Assuntos
Terapia Genética/métodos , Lentivirus/genética , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Fator de Crescimento Neural/genética , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/uso terapêutico , Animais , Arritmias Cardíacas/prevenção & controle , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Cardiotônicos/administração & dosagem , Cardiotônicos/uso terapêutico , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/metabolismo , Injeções Espinhais , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Infarto do Miocárdio/diagnóstico por imagem , Infarto do Miocárdio/prevenção & controle , Fator de Crescimento Neural/biossíntese , Células PC12 , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Canais de Cátion TRPV/antagonistas & inibidores , Canais de Cátion TRPV/metabolismo
3.
Nutrients ; 11(6)2019 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-31167465

RESUMO

Chili peppers are one of the most widely consumed spices worldwide. However, research on the health benefits of chili peppers and some of its constituents has raised controversy as to whether chili pepper compounds possess cancer-promoting or cancer-preventive effects. While ample studies have been carried out to examine the effect of capsaicin in carcinogenesis, the chemopreventive effect of other major components in chili pepper, including dihydrocapsaicin, capsiate, and capsanthin, is relatively unclear. Herein, we investigated the inhibitory effect of chili pepper components on malignant cell transformation. Among the tested chili pepper compounds, dihydrocapsaicin displayed the strongest inhibitory activity against epidermal growth factor (EGF)-induced neoplastic transformation. Dihydrocapsaicin specifically suppressed EGF-induced phosphorylations of the p70S6K1-S6 pathway and the expression of c-Fos. A reduction in c-Fos levels by dihydrocapsaicin led to a concomitant downregulation of AP-1 activation. Further analysis of the molecular mechanism responsible for the dihydrocapsaicin-mediated decrease in phospho-p70S6K1, revealed that dihydrocapsaicin can block amino acid-dependent mechanistic targets of rapamycin complex 1 (mTORC1)-p70S6K1-S6 signal activation. Additionally, dihydrocapsaicin was able to selectively augment amino acid deprivation-induced cell death in mTORC1-hyperactive cells. Collectively, dihydrocapsaicin exerted chemopreventive effects through inhibiting amino acid signaling and c-Fos pathways and, thus, might be a promising cancer preventive natural agent.


Assuntos
Aminoácidos/metabolismo , Capsaicina/análogos & derivados , Células Epiteliais/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Genes fos/fisiologia , Animais , Capsaicina/química , Capsaicina/farmacologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Fator de Crescimento Epidérmico/genética , Fator de Crescimento Epidérmico/metabolismo , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Genes fos/genética , Humanos , Camundongos , Estrutura Molecular , Fosforilação , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Acetato de Tetradecanoilforbol/metabolismo
4.
Bull Exp Biol Med ; 167(1): 43-46, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31177459

RESUMO

We performed a comparative study of the cytotoxic effect of endocannabinoid N-arachidonoyl dopamine (AA-DA) on cultured stromal cells of ectopic and eutopic endometrium. It was found that AA-DA in the concentration range of 1-20 µM produces more selective cytotoxic effect on the stromal cells of the ectopic endometrium due to interaction with cannabinoid type 1 receptor. In concentrations below 1 µM, AA-DA stimulated the proliferation of stromal cells of the eutopic endometrium and did not affect the division of ectopic endometrium cells. This effect was realized due to its interaction with cannabinoid type 2 receptor.


Assuntos
Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Dopamina/metabolismo , Endometriose/metabolismo , Endométrio/metabolismo , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , /farmacologia , Antagonistas de Receptores de Canabinoides/farmacologia , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Endométrio/citologia , Feminino , Humanos , Pirazóis/farmacologia , Receptor CB1 de Canabinoide/antagonistas & inibidores , Receptor CB1 de Canabinoide/metabolismo , Receptor CB2 de Canabinoide/antagonistas & inibidores , Receptor CB2 de Canabinoide/metabolismo , Rimonabanto/farmacologia
5.
J Pharm Biomed Anal ; 173: 126-133, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31129532

RESUMO

A bioanalytical LC-MS/MS method was developed and validated for the simultaneous quantification of capsaicin (CAPS) and dihydrocapsaicin (D-CAPS) in dermal microdialysis samples from rats. Capsaicinoids were separated by using a C18 column, with a mobile phase of water and acetonitrile, both with 0.1% of formic acid, eluted as a gradient. Compounds were detected by using an electrospray ionization source operating in the positive mode (ESI+) to monitor the m/z transitions of 306.1 > 137.0 for CAPS and 308.1 > 137.0 for D-CAPS. The method showed linearity in the concentration range of 0.5-100 ng/ml for CAPS and 0.25-100 ng/ml for D-CAPS, with coefficients of determination of ≥ 0.99. The inter- and intra-day precision, accuracy, and compound stability in different conditions were in accordance with the limits established by the US Food and Drug Administration guidelines. The recovery of the drugs by microdialysis were dependent on the flow rate, but independent of drug concentration. For CAPS, calibration of the in vitro microdialysis probes by dialysis and retrodialysis resulted in statistically similar drug recovery of 68.5% ± 5.9% and 77.8% ± 6.6%, respectively, at a flow rate of 0.5 µl/min. For D-CAPS, the recovery by dialysis was lower than by retrodialysis, at 51.4% ± 6.6% and 92.6% ± 2.4%, respectively. This difference was attributed to the binding of D-CAPS to the plastic tubing, which was experimentally evaluated and mathematically modeled. In vivo recoveries were 75.7% ± 6.3% for CAPS and 81.9% ± 1.5% for D-CAPS at the same flow rate. The analytical method showed high specificity, accuracy, and sensitivity, and suitability for dermatopharmacokinetic studies. These results will allow the determination of the actual free concentration of these drugs in dermatopharmacokinetic experiments, as shown in a pilot experiment with a commercial cream containing capsaicinoids.


Assuntos
Capsaicina/análogos & derivados , Capsaicina/análise , Fármacos do Sistema Sensorial/análise , Creme para a Pele/análise , Animais , Capsaicina/administração & dosagem , Capsaicina/farmacocinética , Cromatografia Líquida de Alta Pressão/métodos , Derme/química , Masculino , Microdiálise/métodos , Modelos Animais , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Fármacos do Sistema Sensorial/administração & dosagem , Fármacos do Sistema Sensorial/farmacocinética , Creme para a Pele/administração & dosagem , Creme para a Pele/farmacocinética , Espectrometria de Massas em Tandem/métodos , Distribuição Tecidual
6.
PLoS One ; 14(4): e0215901, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31039176

RESUMO

ATP-binding cassette (ABC) transporter genes act as transporters for different molecules across biological membranes and are involved in a diverse range of biological processes. In this study, we performed a genome-wide identification and expression analysis of genes encoding ABC transporter proteins in three Capsicum species, i.e., Capsicum annuum, Capsicum baccatum and Capsicum chinense. Capsicum is a valuable horticultural crop worldwide as an important constituent of many foods while containing several medicinal compounds including capsaicin and dihydrocapsaicin. Our results identified the presence of a total of 200, 185 and 187 ABC transporter genes in C. annuum, C. baccatum and C. chinense genomes, respectively. Capsaicin and dihydrocapsaicin content were determined in green pepper fruits (16 dpa). Additionally, we conducted different bioinformatics analyses including ABC genes classification, gene chromosomal location, Cis elements, conserved motifs identification and gene ontology classification, as well as profile expression of selected genes. Based on phylogenetic analysis and domain organization, the Capsicum ABC gene family was grouped into eight subfamilies. Among them, members within the ABCG, ABCB and ABCC subfamilies were the most abundant, while ABCD and ABCE subfamilies were less abundant throughout all species. ABC members within the same subfamily showed similar motif composition. Furthermore, common cis-elements involved in the transcriptional regulation were also identified in the promoter regions of all Capsicum ABC genes. Gene expression data from RNAseq and reverse transcription-semi-quantitative PCR analysis revealed development-specific stage expression profiles in placenta tissues. It suggests that ABC transporters, specifically the ABCC and ABCG subfamilies, may be playing important roles in the transport of secondary metabolites such as capsaicin and dihydrocapsaicin to the placenta vacuoles, effecting on their content in pepper fruits. Our results provide a more comprehensive understanding of ABC transporter gene family in different Capsicum species while allowing the identification of important candidate genes related to capsaicin content for subsequent functional validation.


Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Capsicum/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Proteínas de Plantas/genética , Transportadores de Cassetes de Ligação de ATP/química , Transportadores de Cassetes de Ligação de ATP/metabolismo , Alelos , Motivos de Aminoácidos , Capsaicina/análogos & derivados , Capsaicina/análise , Cromossomos de Plantas/genética , Ontologia Genética , Genes de Plantas , Marcadores Genéticos , Anotação de Sequência Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética
7.
J Agric Food Chem ; 67(22): 6232-6240, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-31075194

RESUMO

This study investigated the effects and molecular mechanism of a combination of capsaicin and capsiate on promoting lipid metabolism and inducing browning in 3T3-L1 white adipocytes. The combination significantly suppressed lipid accumulation in adipocytes ( p = 0.019) and robustly improved lipid metabolic profiles, including decreased triacylglycerol (0.6703 ± 0.0385 versus 0.2849 ± 0.0188 mmol/g of protein; p < 0.001), total cholesterol (0.1282 ± 0.0241 versus 0.0651 ± 0.0178 mmol/g of protein; p = 0.003), and low-density lipoprotein cholesterol (0.0021 ± 0.0017 versus 0.0005 ± 0.0002 mmol/g of protein; p = 0.024) and increased high-density lipoprotein cholesterol (0.0162 ± 0.0141 versus 0.1002 ± 0.0167 mmol/g of protein; p = 0.012). Furthermore, this combination markedly upgraded the protein levels of cluster of differentiation 36 ( p = 0.007) and adipose triglyceride lipase ( p = 0.013) and phosphorylation of hormone-sensitive lipase at Ser660, Ser565, and Ser563 ( p < 0.001, p = 0.027, and p = 0.002, respectively), indicating increases of fatty acid transport and lipolysis. The levels of lipid metabolism regulators, phosphorylation of adenosine-monophosphate-activated protein kinases α and ß ( p = 0.011, and p < 0.001, respectively), sirtuin 1 ( p = 0.004), and vanilloid transient receptor subtype I ( p = 0.014) were also increased by the combination. Moreover, the combination greatly activated the browning program in adipocytes, as demonstrated by increases in beige-specific gene and protein. Further research found that the protein levels of peroxisome proliferator-activated receptor γ (PPARγ; p = 0.001) and ß3-adrenergic receptor (ß3-AR; p = 0.026) were elevated by the combination, and most of the beige-specific markers were abolished by pretreatment of antagonists of PPARγ or ß3-AR. In conclusion, these results indicated that a combination of capsaicin and capsiate could induce browning in white adipocytes via activation of the PPARγ/ß3-AR signaling pathway, and this combination might be worth investigating as a potential cure for obesity.


Assuntos
Adipócitos Marrons/citologia , Adipócitos Brancos/efeitos dos fármacos , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , PPAR gama/metabolismo , Receptores Adrenérgicos beta/metabolismo , Células 3T3-L1 , Adipócitos Marrons/metabolismo , Adipócitos Brancos/citologia , Adipócitos Brancos/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Lipólise/efeitos dos fármacos , Camundongos , PPAR gama/genética , Receptores Adrenérgicos beta/genética , Transdução de Sinais/efeitos dos fármacos
8.
Phytother Res ; 33(7): 1815-1826, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31141276

RESUMO

Capsaicinoid nonivamide (PAVA) and rosuvastatin (RSV) have been shown to exert antioxidant and anti-obesity effects in various animal models, but it is unknown whether their combination would be an effective treatment for obesity-related endothelial dysfunction. This study aimed to investigate the mechanism of PAVA in synergy with RSV. Male Sprague-Dawley rats were given a high-fat diet (HFD) or normal diet during a 20-week period. At 16 weeks, rats in each diet group were divided into subgroups. Normal diet rats were divided into Normal diet control, Normal diet with PAVA, and Normal diet with RSV groups. HFD rats were subdivided into HFD control, HFD with PAVA, HFD with RSV, and HFD with PAVA + RSV groups and evaluated for metabolic parameters, blood pressure, aortic function, and histological change of the aorta in rats. Our results showed the combined therapy had a significantly greater effect than the monotherapy in all measured parameters; this was indicated by improvement in insulin sensitivity and aortic function, decreased blood pressure, lower oxidative stress, and prevention of vascular damage. The synergistic effect of the PAVA and RSV can protect HFD-induced obesity-related endothelial dysfunction, suggesting that the combination of PAVA and RSV could be an effective alternative treatment for obesity-related complications in patients with cardiovascular disease.


Assuntos
Fármacos Antiobesidade/uso terapêutico , Capsaicina/análogos & derivados , Obesidade/tratamento farmacológico , Rosuvastatina Cálcica/uso terapêutico , Animais , Aorta/efeitos dos fármacos , Aorta/fisiologia , Pressão Sanguínea/efeitos dos fármacos , Capsaicina/uso terapêutico , Dieta Hiperlipídica , Sinergismo Farmacológico , Quimioterapia Combinada , Resistência à Insulina , Masculino , Obesidade/fisiopatologia , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley
9.
J Food Sci ; 84(6): 1477-1486, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31132155

RESUMO

Capsicum peppers have not been investigated as sources of quorum sensing (QS) inhibitors. This study aimed to identify compounds in pimenta-malagueta (Capsicum frutescens) and red pepper (Capsicum annuum) extracts and to evaluate their effect on violacein production in Chromobacterium violaceum ATCC 12472 and C. violaceum CV026, as well as biofilm formation (BF) in Pseudomonas aeruginosa PAO1 and Serratia marcescens MG1. Among the extracts, pimenta-malagueta methanolic extract (PMME) was chosen because it contained capsaicin, dihydrocapsaicin, and luteolin in greater amount than the other extracts. In general, PMME partially inhibited bacterial growth at 2.5 and 5.0 mg/mL, as well as capsaicin at 100 µg/mL and luteolin at 62.5, 125, and 250 µg/mL. At lower concentrations, PMME and luteolin reduced violacein production in C. violaceum ATCC 12472 without affecting growth, a result that was not observed with capsaicin. We show that violacein inhibition by PMME is likely due to luteolin. In silico docking evaluation showed that luteolin binds to the CviR QS regulator. Crystal violet staining and confocal microscopy revealed that BF was increased by PMME and capsaicin, being remarkably superior for P. aeruginosa PAO1 at 30 °C. Capsaicin is not an effective QS inhibitor, while luteolin should be further investigated for its potential effects in QS regulated phenotypes. PRACTICAL APPLICATION: Quorum sensing (QS) is a form of bacterial communication targeted for studies aiming to inhibit bacterial virulence. QS regulates phenotypes that influence microbial activities across many areas, including Food Science. Capsicum frutescens is a type of chili pepper consumed in Brazil, rich in bioactive compounds such as capsaicin (which gives its pungency) and luteolin (a phenolic compound). We show that C. frutescens extract and luteolin inhibit QS in a model bacterium, along with the possible molecular mechanism of inhibition. Capsaicin did not inhibit QS neither biofilm formation. Luteolin should be further investigated for its QS inhibition properties and biotechnological applications.


Assuntos
Capsaicina/farmacologia , Capsicum/química , Luteolina/farmacologia , Extratos Vegetais/farmacologia , Percepção de Quorum/efeitos dos fármacos , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Brasil , Capsaicina/análogos & derivados , Capsaicina/análise , Chromobacterium/efeitos dos fármacos , Frutas/química , Luteolina/análise , Fenótipo , Extratos Vegetais/química , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/fisiologia , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/fisiologia
10.
Nutrients ; 11(4)2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-31022842

RESUMO

In this study, two capsaicin analogues, N-eicosapentaenoyl vanillylamine (EPVA) and N-docosahexaenoyl vanillylamine (DHVA), were enzymatically synthesized from their corresponding n-3 long chain polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), both dietary relevant components. The compounds significantly reduced the production of some lipopolysaccharide (LPS)-induced inflammatory mediators, including nitric oxide (NO), macrophage-inflammatory protein-3α (CCL20) and monocyte chemoattractant protein-1 (MCP-1 or CCL2), by RAW264.7 macrophages. Next to this, only EPVA increased insulin secretion by pancreatic INS-1 832/13 ß-cells, while raising intracellular Ca2+ and ATP concentrations. This suggests that the stimulation of insulin release occurs through an increase in the intracellular ATP/ADP ratio in the first phase, while is calcium-mediated in the second phase. Although it is not yet known whether EPVA is endogenously produced, its potential therapeutic value for diabetes treatment merits further investigation.


Assuntos
Capsaicina/análogos & derivados , Capsaicina/farmacologia , Ácidos Docosa-Hexaenoicos/análogos & derivados , Ácido Eicosapentaenoico/análogos & derivados , Ácidos Graxos Ômega-3/química , Inflamação/metabolismo , Insulina/metabolismo , Macrófagos/efeitos dos fármacos , Animais , Cálcio/metabolismo , Capsaicina/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ácidos Docosa-Hexaenoicos/síntese química , Ácidos Docosa-Hexaenoicos/química , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/síntese química , Ácido Eicosapentaenoico/química , Ácido Eicosapentaenoico/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/metabolismo , Macrófagos/metabolismo , Camundongos , Estrutura Molecular , Células RAW 264.7 , Ratos
11.
J Oral Pathol Med ; 48(5): 389-399, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30825343

RESUMO

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a deadly disease with a mere 40% five-year survival rate for patients with advanced disease. Previously, we discovered that capsazepine (CPZ), a transient receptor potential channel, Vanilloid subtype 1 (TRPV1) antagonist, has significant anti-tumor effects against OSCC via a unique mechanism-of-action that is independent of TRPV1. Thus, we developed novel CPZ analogs with more potent anti-proliferative effects (CIDD-24, CIDD-99, and CIDD-111). METHODS: Using OSCC xenograft models, we determined the efficacy of these analogs in vivo. TRPV1 interactions were evaluated using calcium imaging and a rat model of orofacial pain. Anti-cancer mechanism(s)-of-action were assessed by cell cycle analysis and mitochondrial depolarization assays. RESULTS: CIDD-99 was the most potent analog demonstrating significant anti-tumor effects in vivo (P < 0.001). CIDD-24 was equipotent to the parent compound CPZ, but less potent than CIDD-99. CIDD-111 was the least efficacious analog. Calcium imaging studies confirmed that CIDD-99 neither activates nor inhibits TRPV1 confirming that TRPV1 activity is not involved in its anti-cancer effects. All analogs induced an S-phase block, dose-dependent mitochondrial depolarization, and apoptosis. Histological analyses revealed increased apoptosis and reduced cell proliferation in tumors treated with these analogs. Importantly, CIDD-99 had the most dramatic anti-tumor effects with 85% of tumors resolving leaving only minute traces of viable tissue. Additionally, CIDD-99 was non-noxious and demonstrated no observable adverse reactions CONCLUSION: This study describes a novel, highly efficacious, CPZ analog, CIDD-99, with dramatic anti-tumor effects against OSCC that may be efficacious as a lone therapy or in combination with standard therapies.


Assuntos
Apoptose , Capsaicina/análogos & derivados , Carcinoma de Células Escamosas/tratamento farmacológico , Estresse do Retículo Endoplasmático , Isoquinolinas/farmacologia , Mitocôndrias/efeitos dos fármacos , Neoplasias Bucais/tratamento farmacológico , Canais de Cátion TRPV/antagonistas & inibidores , Animais , Capsaicina/farmacologia , Linhagem Celular Tumoral , Feminino , Humanos , Masculino , Camundongos , Camundongos Nus , Mitocôndrias/patologia , Ratos , Ratos Sprague-Dawley , Ensaios Antitumorais Modelo de Xenoenxerto
12.
Molecules ; 24(5)2019 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-30871017

RESUMO

Capsazepine is a synthetic analogue of capsaicin that can function as an antagonist of TRPV1. Capsazepine can exhibit diverse effects on cancer (prostate cancer, breast cancer, colorectal cancer, oral cancer, and osteosarcoma) growth and survival, and can be therapeutically used against other major disorders such as colitis, pancreatitis, malaria, and epilepsy. Capsazepine has been reported to exhibit pleiotropic anti-cancer effects against numerous tumor cell lines. Capsazepine can modulate Janus activated kinase (JAK)/signal transducer and activator of the transcription (STAT) pathway, intracellular Ca2+ concentration, and reactive oxygen species (ROS)-JNK-CCAAT/enhancer-binding protein homologous protein (CHOP) pathways. It can inhibit cell proliferation, metastasis, and induce apoptosis. Moreover, capsazepine can exert anti-inflammatory effects through the downregulation of lipopolysaccharide (LPS)-induced nuclear transcription factor-kappa B (NF-κB), as well as the blockage of activation of both transient receptor potential cation channel subfamily V member 1 (TRPV1) and transient receptor potential cation channel, subfamily A, and member 1 (TRPA1). This review briefly summarizes the diverse pharmacological actions of capsazepine against various cancers and inflammatory conditions.


Assuntos
Anti-Inflamatórios/farmacologia , Antineoplásicos/farmacologia , Capsaicina/análogos & derivados , Animais , Capsaicina/farmacologia , Linhagem Celular Tumoral , Humanos , Janus Quinases/antagonistas & inibidores , NF-kappa B/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Canais de Cátion TRPV/efeitos dos fármacos
13.
Theriogenology ; 128: 207-217, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30784807

RESUMO

In view of the limited information available on functional significance of TRPV1 in regulating sperm functions, present study was undertaken on bull spermatozoa. Sixty four ejaculates were collected from four Hariana bulls and were used for molecular and functional characterisation of TRPV1. Immunoblotting using TRPV1 specific antibody revealed the presence of a single band of 104 kDa corresponding to TRPV1 in Hariana bull spermatozoa. Indirect immuno fluorescence revealed positive immune-reactivity to TRPV1 at acrosomal, pre-acrosomal, post acrosomal and flagellar regions of spermatozoa. Based on the results of pilot study dose-response analysis, doses of anandamide (AEA; 0.3 µM) and capsazepine (Cp; 10 µM) were selected for further studies. Three groups of semen samples (control 100 µL diluted semen having 1 × 106 spermatozoa; anandamide (3 µL AEA+97 µL of diluted semen containing 1 × 106 spermatozoa and Cp (1 µL Cp+99 µL of diluted semen containing 1 × 106 spermatozoa) were used to study the functional involvement of TRPV1 in bull spermatozoa. Blocking of TRPV1 with Cp resulted in significant (P < 0.05) reduction in progressive sperm motility (PSM) as compared to control. With activation of TRPV1 using AEA also, PSM was significantly (P < 0.05) decreased till 1h and thereafter the PSM was sustained to the level as observed in control. However, both during blocking and activation of TRPV1, per cent spermatozoa showing hyperactive motility were significantly (P < 0.05) increased (20-30%) compared to the control. Treatment with both Cp and AEA revealed significant (P < 0.05) increase in B-pattern of spermatozoa in chlortetracycline hydrochloride (CTC) staining indicating induction of capacitation. Inhibition of soluble adenyl cyclase (sAC) with 99 nM KH7and protein kinase A (PKA) with 3 µM P9115 significantly (P < 0.05) decreased PSM both in the presence of Cp and AEA. Blocking as well as activation of TRPV1 showed significant (P < 0.05) reduction in sperm livability, intact membrane, intact acrosome, high mitochondrial transmembrane potential; hence indicating the involvement of TRPV1 in regulation of sperm functions in bulls. From the study-it was concluded that TRPV1 channels are found in bull spermatozoa and mediate number of sperm functions like motility, hypermotility, capacitation and acrosome reaction. Further studies are required to find out the possible relationship between TRPV1 channels and other channels in regulating spermatozoa function and possible mechanisms associated with TRPV1 activation as well as its role in sperm function regulation.


Assuntos
Espermatozoides/fisiologia , Canais de Cátion TRPV/fisiologia , Reação Acrossômica , Animais , Ácidos Araquidônicos/farmacologia , Bloqueadores dos Canais de Cálcio/farmacologia , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Bovinos , Endocanabinoides/farmacologia , Masculino , Alcamidas Poli-Insaturadas/farmacologia , Capacitação Espermática , Motilidade Espermática , Espermatozoides/metabolismo , Canais de Cátion TRPV/metabolismo
14.
PLoS One ; 14(1): e0210510, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30673734

RESUMO

Capsaicinoids are compounds synthesized exclusively in the genus Capsicum and are responsible for the burning sensation experienced when consuming hot pepper fruits. To date, only one gene, AT3, a member of the BAHD family of acyltransferases, is currently known to have a measurable quantitative effect on capsaicinoid biosynthesis. Multiple AT3 paralogs exist in the Capsicum genome, but their evolutionary relationships have not been characterized well. Recessive alleles at this locus result in absence of capsaicinoids in pepper fruit. To explore the evolution of AT3 in Capsicum and the Solanaceae, we sequenced this gene from diverse Capsicum genotypes and species, along with a number of representative solanaceous taxa. Our results revealed that the coding region of AT3 is highly conserved throughout the family. Further, we uncovered a tandem duplication that predates the diversification of the Solanaceae taxa sampled in this study. This pair of tandem duplications were designated AT3-1 and AT3-2. Sequence alignments showed that the AT3-2 locus, a pseudogene, retains regions of amino acid conservation relative to AT3-1. Gene tree estimation demonstrated that AT3-1 and AT3-2 form well supported, distinct clades. In C. rhomboideum, a non-pungent basal Capsicum species, we describe a recombination event between AT3-1 and AT3-2 that modified the putative active site of AT3-1, also resulting in a frame-shift mutation in the second exon. Our data suggest that duplication of the original AT3 representative, in combination with divergence and pseudogene degeneration, may account for the patterns of sequence divergence and punctuated amino acid conservation observed in this study. Further, an early rearrangement in C. rhomboidium could account for the absence of pungency in this Capsicum species.


Assuntos
Capsaicina/análogos & derivados , Capsicum/genética , Duplicação Gênica , Genes de Plantas/genética , Recombinação Genética , Solanaceae/genética , Aciltransferases/genética , Aciltransferases/metabolismo , Sequência de Bases , Capsaicina/metabolismo , Capsicum/classificação , Capsicum/metabolismo , Filogenia , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Solanaceae/classificação , Solanaceae/metabolismo , Especificidade da Espécie
15.
Rapid Commun Mass Spectrom ; 33(7): 635-640, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30668887

RESUMO

RATIONALE: Capsaicinoids are prevalent secondary metabolites in many natural and synthetic pharmacological compounds. To date, several soft ionization studies related to capsaicinoids have been reported; they all proposed a common fragmentation pattern based on a rearrangement of the aromatic double bonds and the fragmentation of the various positional acyl chains. However, the mechanism has never been validated by high-resolution analyses. Consequently, in this work, a validated fragmentation mechanism of the main capsaicinoids, capsaicin (1) and dihydrocapsaicin (2), is offered. METHODS: In order to propose and validate a common electron ionization (EI) fragmentation mechanism for the target analytes, the following mass spectrometric methods were employed: collision-induced dissociation (CID) by means of linked scans (LS), reinforcing the methodology by high-resolution mass spectrometry (HRMS), in addition to appropriate deuterium-labeled experiments performed using gas chromatography/mass spectrometry (GC/MS) and direct analysis in real time (DART). RESULTS: In a first stage, a common EI fragmentation pattern comprising two pathways was proposed for compounds 1 and 2; then, the suggested mechanism was validated by CID-LS together with HRMS complemented by DART-deuterium-labeling studies. The obtained results are indicative that the corresponding molecular ions were conveniently observed, m/z 305 and m/z 307; it is worth noting that the common base peak is in correspondence with a tropylium ion derivative (m/z 137), as a consequence of a McLafferty rearrangement. In addition to these highlighted fragments, other common ions, m/z 122 and m/z 94, and their corresponding trajectory, were confirmed using the same approach. Finally, the proposed mechanism was complementarily validated by deuterium-labeling studies, taking into account the two exchangeable hydrogens present in the phenolic and the amidic moieties. CONCLUSIONS: A common validated EI fragmentation pattern for both capsaicin and dihydrocapsaicin was established using appropriated mass spectrometric methods together with convenient hydrogen/deuterium labeling. This study provides a new alternative to validate mechanisms of fragmentation of important natural products.


Assuntos
Capsaicina/análogos & derivados , Capsaicina/química , Espectrometria de Massas/métodos , Capsaicina/análise , Capsicum/química , Medição da Troca de Deutério , Cromatografia Gasosa-Espectrometria de Massas/métodos , Íons/análise , Íons/química , Reprodutibilidade dos Testes
16.
Bull Exp Biol Med ; 166(3): 310-312, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30627906

RESUMO

Application of mild irritants (1 M NaCl; pH 2.0) on the gastric mucosa potentiates the protective secretion of bicarbonates by epithelial cells. This response is mainly mediated by capsaicin-sensitive afferent nerve endings located in the submucosa. It was shown that activation of vanilloid type 1 receptors (TRPV1) induced by exogenous acidification of GM is not sufficient to potentiate the production of HCO3, including production depending on neuronal NO synthase. However, the effect of exogenous acid on TRPV1 leads to activation of endothelial NO synthase that restrict the gastric secretion of [Formula: see text].


Assuntos
Bicarbonatos/metabolismo , Mucosa Gástrica/efeitos dos fármacos , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo I/genética , Cloreto de Sódio/farmacologia , Estômago/efeitos dos fármacos , Canais de Cátion TRPV/genética , Amilorida/farmacologia , Animais , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Mucosa Gástrica/inervação , Mucosa Gástrica/metabolismo , Regulação da Expressão Gênica , Concentração de Íons de Hidrogênio , Indazóis/farmacologia , Masculino , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I/metabolismo , Óxido Nítrico Sintase Tipo III/antagonistas & inibidores , Óxido Nítrico Sintase Tipo III/metabolismo , Nitroarginina/farmacologia , Concentração Osmolar , Perfusão , Nervo Frênico/cirurgia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Estômago/inervação , Canais de Cátion TRPV/antagonistas & inibidores , Canais de Cátion TRPV/metabolismo , Vagotomia
17.
Biosens Bioelectron ; 126: 815-823, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30602263

RESUMO

In this study, a taste bud tissue biosensor was prepared by a starch-sodium alginate cross-linking fixation method. Capsaicin was used as a TRPV1 noxious ion channel activator to investigate the antagonism kinetics of six different substances on capsaicin. The results showed that capsazepine, AMG517, loureirin B, and tetrahydropalmatine were all competitive allosteric regulatory ligands for capsaicin, while aconitine and anandamide were mixed allosteric regulatory ligand that combines non-competition and competition effect. Through analyzing the kinetic parameters of capsaicin and its competitive allosteric regulatory ligands, and comparing the structures between spicy substances and endocannabinoids, the importance of amide groups and similar groups in the allosteric regulation of cannabinoids (CB) receptors and analgesic mechanism was elucidated. This indicates that vanilloid activators turn on the TRPV1 ion channel to transmit only pain and other nociceptive signals, while capsaicin and its competitive ligands are capable of activating intracellular G protein/PI3K/PIP2 signaling pathways by binding to endogenous cannabinoid receptors, and then increase intracellular PIP2 levels (the increasing PIP2 can competitively replace capsaicin and other vanilloid activators), thereby closing the TRPV1 channel and exerting the analgesic effect. The elucidation of this mechanism of pain and analgesia will lay the theoretical foundation and new ideas for investigating nociceptive signal and screening potential analgesic drugs.


Assuntos
Analgésicos/farmacologia , Técnicas Biossensoriais/métodos , Papilas Gustativas/efeitos dos fármacos , Alginatos/química , Regulação Alostérica , Analgésicos/efeitos adversos , Animais , Ácidos Araquidônicos/metabolismo , Técnicas Biossensoriais/instrumentação , Temperatura Corporal/efeitos dos fármacos , Capsaicina/análogos & derivados , Capsaicina/metabolismo , Capsaicina/farmacologia , Reagentes para Ligações Cruzadas/química , Técnicas Eletroquímicas/métodos , Endocanabinoides/metabolismo , Febre/induzido quimicamente , Dor/tratamento farmacológico , Dor/metabolismo , Dor/fisiopatologia , Alcamidas Poli-Insaturadas/metabolismo , Ratos Sprague-Dawley , Amido/química , Canais de Cátion TRPV/metabolismo , Papilas Gustativas/fisiologia
18.
BMC Neurosci ; 20(1): 1, 2019 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-30602386

RESUMO

BACKGROUND: Peripheral diabetic neuropathy can be painful and its symptoms include hyperalgesia, allodynia and spontaneous pain. Hydrogen sulfide (H2S) is involved in diabetes-induced hyperalgesia and allodynia. However, the molecular target through which H2S induces hyperalgesia in diabetic animals is unclear. The aim of this study was to determine the possible involvement of transient receptor potential (TRP) channels in H2S-induced hyperalgesia in diabetic rats. RESULTS: Streptozotocin (STZ) injection produced hyperglycemia in rats. Intraplantar injection of NaHS (an exogenous donor of H2S, 3-100 µg/paw) induced hyperalgesia, in a time-dependent manner, in formalin-treated diabetic rats. NaHS-induced hyperalgesia was partially prevented by local intraplantar injection of capsazepine (0.3-3 µg/paw), HC-030031 (100-316 µg/paw) and SKF-96365 (10-30 µg/paw) blockers, at 21 days post-STZ injection. At the doses used, these blockers did not modify formalin-induced nociception. Moreover, capsazepine (0.3-30 µg/paw), HC-030031 (100-1000 µg/paw) and SKF-96365 (10-100 µg/paw) reduced formalin-induced nociception in diabetic rats. Contralateral injection of the highest doses used did not modify formalin-induced flinching behavior. Hyperglycemia, at 21 days, also increased protein expression of cystathionine-ß-synthase enzyme (CBS) and TRPC6, but not TRPA1 nor TRPV1, channels in dorsal root ganglia (DRG). Repeated injection of NaHS enhanced CBS and TRPC6 expression, but hydroxylamine (HA) prevented the STZ-induced increase of CBS protein. In addition, daily administration of SKF-96365 diminished TRPC6 protein expression, whereas NaHS partially prevented the decrease of SKF-96365-induced TRPC6 expression. Concordantly, daily intraplantar injection of NaHS enhanced, and HA prevented STZ-induced intraepidermal fiber loss, respectively. CBS was expressed in small- and medium-sized cells of DRG and co-localized with TRPV1, TRPA1 and TRPC6 in IB4-positive neurons. CONCLUSIONS: Our data suggest that H2S leads to hyperalgesia in diabetic rats through activation of TRPV1, TRPA1 and TRPC channels and, subsequent intraepidermal fibers loss. CBS enzyme inhibitors or TRP-channel blockers could be useful for treatment of painful diabetic neuropathy.


Assuntos
Diabetes Mellitus Experimental/metabolismo , Sulfeto de Hidrogênio/metabolismo , Hiperalgesia/metabolismo , Canais de Receptores Transientes de Potencial/metabolismo , Acetanilidas/farmacologia , Analgésicos/farmacologia , Animais , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Cistationina beta-Sintase/metabolismo , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/patologia , Feminino , Formaldeído , Hidroxilamina/farmacologia , Hiperalgesia/tratamento farmacológico , Hiperalgesia/patologia , Imidazóis/farmacologia , Nociceptividade/efeitos dos fármacos , Nociceptividade/fisiologia , Purinas/farmacologia , Ratos Wistar , Pele/inervação , Pele/metabolismo , Raízes Nervosas Espinhais/efeitos dos fármacos , Raízes Nervosas Espinhais/metabolismo , Raízes Nervosas Espinhais/patologia , Sulfitos
19.
J Sci Food Agric ; 99(1): 269-280, 2019 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-29851100

RESUMO

BACKGROUND: Assessment of the different desirable characters among chili genotypes has expanded the effective selection for crop improvement. Identification of genetically superior parents is important in assortment of the best parents to develop new chili hybrids. RESULTS: This study was done to assess the hereditary assorted variety of selected genotypes of Capsicum annuum based on their morphophysiological and yield traits in two planting seasons. The biochemical properties, capsaicinoid content (capsaicin and dihydrocapsaicin), total phenolics content and antioxidant action determination of unripe and ripe chili pepper fruits were carried out in dry fruits. AVPP9813 and Kulai 907 were observed to have high fruit yields, with 541.39 and 502.64 g per plant, respectively. The most increased genotypic coefficient of variation (GCV) and phenotypic coefficient of variation (PCV) were shown by the fruit number per plant (49.71% and 66.04%, respectively). High heritability was observed in yield characters viz-à-viz fruit weight, length and girth and indicated high genetic advance. Eight groups were obtained from the cluster analysis. For the biochemical analysis, the capsaicinoid content and total phenolic content were high in Chili Bangi 3 at unripe and ripe fruit stages, while for antioxidant activity SDP203 was the highest in ripe dry fruit. CONCLUSION: Higher GCV and PCV, combined with moderate to high heritability and high hereditary progress, were seen in number of fruit per plant, fruit yield per plant and fruit weight per fruit. These findings are beneficial for chili pepper breeders to select desirable quantitative characters in C. annuum in their breeding program. © 2018 Society of Chemical Industry.


Assuntos
Capsicum/genética , Frutas/química , Frutas/crescimento & desenvolvimento , Variação Genética , Capsaicina/análogos & derivados , Capsaicina/análise , Capsicum/química , Capsicum/classificação , Capsicum/crescimento & desenvolvimento , Frutas/classificação , Frutas/genética , Genótipo , Fenóis/análise , Extratos Vegetais/análise
20.
Bioorg Med Chem ; 27(1): 208-215, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30528162

RESUMO

We previously demonstrated that capsazepine (CPZ), a synthetic transient receptor potential Vanilloid subtype 1 (TRPV1) antagonist, has significant anti-cancer effects in vivo. The purpose of this study was to develop more potent analogs based upon CPZ pharmacophore and structure-activity relationships (SAR) across analogs. We generated 30 novel compounds and screened for their anti-proliferative effects in cultured HeLa cervical cancer cells. Cell viability assays identified multiple compounds with IC50s < 15 µM and one compound, 29 with an IC50 < 5 µM; six fold more potent than CPZ. We validated the anti-proliferative efficacy of two lead compounds, 17 and 29, in vivo using HeLa-derived xenografts in athymic nude mice. Both analogs significantly reduced tumor volumes by day 8 compared to control treated animals (p < 0.001) with no observable adverse effects. Calcium imaging determined that compound 17 activates TRPV1 whereas 29 neither activates nor inhibits TRPV1; indicating a unique mechanism-of-action that does not involve TRPV1 signaling. Cell viability assays using a panel of additional tumor types including oral squamous cell carcinoma, non-small cell lung cancer (NSCLC), breast cancer, and prostate cancer cell lines (HSC-3, H460, MDA-231, and PC-3 respectively) demonstrated that both lead compounds were efficacious against every cancer type tested. Compounds 29 displayed IC50s of 1-2.5 µM in HSC-3and PC-3cells. Thus, we propose that these novel CPZ analogs may serve as efficacious therapeutic agents against multiple tumor types that warrant further development for clinical application.


Assuntos
Antineoplásicos/uso terapêutico , Capsaicina/análogos & derivados , Neoplasias/tratamento farmacológico , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Capsaicina/síntese química , Capsaicina/farmacologia , Capsaicina/uso terapêutico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Humanos , Camundongos Nus , Estrutura Molecular , Relação Estrutura-Atividade , Canais de Cátion TRPV/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
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