RESUMO
Arthritis has important cardiovascular repercussions. Phenylephrine-induced vasoconstriction is impaired in rat aortas in the early phase of the adjuvant-induced arthritis (AIA), around the 15th day post-induction. Therefore, the present study aimed to verify the effects of AIA on hyporesponsiveness to phenylephrine in rat aortas. AIA was induced by intradermal injection of Mycobacterium tuberculosis (3.8 mg/dL) in the right hind paw of male Wistar rats (n=27). Functional experiments in isolated aortas were carried out 15 days after AIA induction. Morphometric and stereological analyses of the aortas were also performed 36 days after the induction of AIA. AIA did not promote structural modifications in the aortas at any of the time points studied. AIA reduced phenylephrine-induced contraction in endothelium-intact aortas, but not in endothelium-denuded aortas. However, AIA did not change KCl-induced contraction in either endothelium-intact or denuded aortas. L-NAME (non-selective NOS inhibitor), 1400W (selective iNOS inhibitor), and ODQ (guanylyl cyclase inhibitor) reversed AIA-induced hyporesponsiveness to phenylephrine in intact aortas. 7-NI (selective nNOS inhibitor) increased the contraction induced by phenylephrine in aortas from AIA rats. In summary, the hyporesponsiveness to phenylephrine induced by AIA was endothelium-dependent and mediated by iNOS-derived NO through activation of the NO-guanylyl cyclase pathway.
Assuntos
Artrite Experimental , Óxido Nítrico , Fenilefrina , Ratos Wistar , Animais , Masculino , Fenilefrina/farmacologia , Artrite Experimental/fisiopatologia , Artrite Experimental/induzido quimicamente , Óxido Nítrico/metabolismo , Vasoconstrição/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Vasoconstritores/farmacologia , Ratos , Aorta/efeitos dos fármacosRESUMO
Arthritis has important cardiovascular repercussions. Phenylephrine-induced vasoconstriction is impaired in rat aortas in the early phase of the adjuvant-induced arthritis (AIA), around the 15th day post-induction. Therefore, the present study aimed to verify the effects of AIA on hyporesponsiveness to phenylephrine in rat aortas. AIA was induced by intradermal injection of Mycobacterium tuberculosis (3.8 mg/dL) in the right hind paw of male Wistar rats (n=27). Functional experiments in isolated aortas were carried out 15 days after AIA induction. Morphometric and stereological analyses of the aortas were also performed 36 days after the induction of AIA. AIA did not promote structural modifications in the aortas at any of the time points studied. AIA reduced phenylephrine-induced contraction in endothelium-intact aortas, but not in endothelium-denuded aortas. However, AIA did not change KCl-induced contraction in either endothelium-intact or denuded aortas. L-NAME (non-selective NOS inhibitor), 1400W (selective iNOS inhibitor), and ODQ (guanylyl cyclase inhibitor) reversed AIA-induced hyporesponsiveness to phenylephrine in intact aortas. 7-NI (selective nNOS inhibitor) increased the contraction induced by phenylephrine in aortas from AIA rats. In summary, the hyporesponsiveness to phenylephrine induced by AIA was endothelium-dependent and mediated by iNOS-derived NO through activation of the NO-guanylyl cyclase pathway.
RESUMO
Probiotics have aroused great interest as an adjunctive treatment to periodontal therapy, due to the frequent colonisation by periodontopathogens after therapy. The aim of this systematic review was to analyse in the scientific literature, evidence of the microbiological effects of probiotics as an adjunct to periodontal therapy in the treatment of periodontal diseases (PD). Only randomised controlled trials (RCT), evaluating the microbiological effect of probiotics as an adjunct to periodontal therapy. The authors conducted a search in PubMed/MEDLINE, LILACS, ScienceDirect, Web of Science and Cochrane Library to identify articles published in English until February 2020. The quality of the studies was assessed using the JADAD scale and the risk of bias was assessed according to the Cochrane Collaboration assessment tool. Of the 265 articles potentially relevant to this review, 10 studies were included. The most frequently used probiotic bacteria were those of the genus Lactobacillus spp. and the time of administration of the probiotics was between 14 days to 3 months. Most studies have shown that the adjuvant use of probiotics reduces the total mean counts of gram-negative anaerobic species (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Prevotella intermedia) and gram-negative coccobacillus (Aggregatibacter actinomycetemcomitans) of subgingival plaque samples. Probiotics adjuvant to periodontal therapy reduces periodontopathogenic species in a greater proportion, compared only to periodontal therapy. Especially the Lactobacillus reuteri strain, without combination with other strains, offered a greater reduction in pathogenic bacteria associated with greater destruction of periodontal tissues and deep periodontal pockets. Researchers should perform high-quality RCT, evaluating single strains without combinations, in order to observe the microbiological benefits as adjunctive treatment of PD.
Assuntos
Doenças Periodontais , Probióticos , Humanos , Limosilactobacillus reuteri , Doenças Periodontais/terapia , Bolsa Periodontal/microbiologia , Probióticos/uso terapêutico , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Objetivo. El objetivo de este estudio fue evaluar la adaptación marginal de coronas de disilicato de litio obtenidas mediante técnicas de escaneo (CAD/CAM), antes y des- pués de la cristalización, a través de análisis in vitro con microscopía confocal (MC). Métodos. Fueron confeccionadas 16 réplicas en poliuretano a partir de la pieza 1.4, de modelo typodont, tallada para corona total. Las réplicas fueron divididas en dos gru- pos, de acuerdo a la técnica de escaneo: Técnica Indirecta (Grupo IND, n=08), donde modelos de yeso fueron escaneados con escáner de laboratorio (inEos X5, Sirona Den- tal Systems) y Técnica Directa (Grupo DIR, n=08), donde modelos typodont fueron escaneados con escáner intraoral (CEREC BlueCam, Sirona Dental Systems). A seguir, se fresaron (inLab MC XL, Sirona Dental Systems) coronas en disilicato de litio (IPS e.max CAD, Ivoclar Vivadent) y se adaptaron a las réplicas. Se evaluó la adaptación marginal con análisis de MC en dos momentos, antes y después de la cristalización del disilicato de litio. Los datos fueron analizados con la prueba de Mann-Whitney, t de Student y Wilcoxon (α= 0,05). Resultados. Hubo una diferencia estadísticamente significativa en la adaptación marginal horizontal entre los grupos IND y DIR después de la cristalización (p=0,05). En el grupo IND, la comparación de la adaptación mar- ginal vertical antes y después de la cristalización mostró una diferencia estadísticamente significativa (p=0,038). Conclusiones. Las coronas de disilicato de litio obtenidas me- diante escaneo directo (CAD/CAM) presentaron menor desajuste marginal vertical. La etapa de cristalización afectó la adaptación marginal de las coronas.
Objective. This study aimed to evaluate lithium disilicate marginal adaption on crowns by scanning techniques (CAD/CAM), before and after crystallization, through confocal microscopy (CM) in vitro analysis. Methods. Sixteen polyurethane replicas were per- formed from tooth 1.4, of a typodont model, prepared for a full crown. The replicas were divided into two groups, according to the scanning technique: Indirect Technique (Group IND, n=08), where dental stone models were scanned with a laboratory scanner (inEos X5, Sirona Dental Systems) and Direct Technique (Group DIR, n=08), where typodont models were scanned with an intraoral scanner (CEREC BlueCam, Sirona Dental Systems). Then, the lithium disilicate crowns (IPS e.max CAD, Ivoclar Vivadent) were milled (inLab MC XL, Sirona Dental Systems) and adapted to the replicas. Margin- al adaptation was evaluated with CM analysis before and after lithium disilicate crystalli- zation. Data were analyzed with the Mann-Whitney, t test, and Wilconxon test (α=0.05). Results. There was a statistically significant difference in horizontal marginal adaptation between IND and DIR groups after crystallization (p=0.05). In IND group, the compar- ison of vertical marginal adaptation before and after crystallization showed a statistically significant difference (p=0.038). Conclusions. Lithium disilicate crowns obtained by direct scanning technique (CAD/CAD) showed less vertical marginal maladjustment. The crystallization stage affected the crown's marginal adaptation.
RESUMO
AIMS: We determined whether decreased reactive oxygen species (ROS) production in the aorta of pregnant spontaneously hypertensive rats (SHR) resulted in increased nitric oxide (NO) bioavailability and hyporeactivity to phenylephrine (PE). MAIN METHODS: Systemic and aortic oxidative stress were measured in pregnant and non-pregnant Wistar rats and SHR. Furthermore, the hypotensive effects of apocynin (30 mg/kg) and Tempol (30 mg/kg) were analyzed. Intact aortic rings of pregnant and non-pregnant rats were stimulated with PE in the absence of or after incubation (30 min) with apocynin (100 µmol/L). The effect of apocynin on the concentrations of NO and ROS were measured in aortic endothelial cells (AEC) using DAF-2DA (10 mmol/L) and DHE (2.5 mmol/L), respectively. Western blotting was performed to analyze eNOS, NOX1, NOX2, NOX4 and SOD expression. ROS production was analyzed by the lucigenin chemiluminescence method. KEY FINDINGS: Aortic oxidative stress and ROS concentration in AEC were reduced in pregnant Wistar rats and SHR, when compared to non-pregnant rats. ROS production and NOX1, NOX2 and NOX4 expression in the aortas were decreased in pregnant SHR, but not in pregnant Wistar rats. Increased eNOS expression in aortas and NO concentration in AEC were observed in pregnant Wistar rats and SHR. Apocynin reduced PE-induced vasoconstriction in the aortas of non-pregnant Wistar rats and SHR, and pregnant Wistar rats, but not in the aortas of pregnant SHR. SIGNIFICANCE: Taken together, these results suggest that ROS production was decreased in the aortas of pregnant SHR and could contribute to higher NO bioavailability and hyporeactivity to PE in the aortas of pregnant SHR.
Assuntos
Aorta Torácica/enzimologia , Cardiotônicos/farmacologia , Glicoproteínas de Membrana/biossíntese , NADH NADPH Oxirredutases/biossíntese , NADPH Oxidases/biossíntese , Fenilefrina/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Anti-Hipertensivos/farmacologia , Aorta Torácica/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Feminino , Frequência Cardíaca/efeitos dos fármacos , Técnicas In Vitro , NADPH Oxidase 1 , NADPH Oxidase 2 , NADPH Oxidase 4 , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Gravidez , Ratos , Ratos Endogâmicos SHR , Ratos Wistar , Vasoconstrição/efeitos dos fármacosRESUMO
The aim of the present study was to determine the mechanisms underlying the relaxant effect of adrenomedullin (AM) in rat cavernosal smooth muscle (CSM) and the expression of AM system components in this tissue. Functional assays using standard muscle bath procedures were performed in CSM isolated from male Wistar rats. Protein and mRNA levels of pre-pro-AM, calcitonin receptor-like receptor (CRLR), and Subtypes 1, 2 and 3 of the receptor activity-modifying protein (RAMP) family were assessed by Western immunoblotting and quantitative real-time polymerase chain reaction, respectively. Nitrate and 6-keto-prostaglandin F1α (6-keto-PGF1α; a stable product of prostacyclin) levels were determined using commercially available kits. Protein and mRNA of AM, CRLR, and RAMP 1, -2, and -3 were detected in rat CSM. Immunohistochemical assays demonstrated that AM and CRLR were expressed in rat CSM. AM relaxed CSM strips in a concentration-dependent manner. AM22-52, a selective antagonist for AM receptors, reduced the relaxation induced by AM. Conversely, CGRP8-37, a selective antagonist for calcitonin gene-related peptide receptors, did not affect AM-induced relaxation. Preincubation of CSM strips with NG-nitro-L-arginine-methyl-ester (L-NAME, nitric oxide synthase inhibitor), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, quanylyl cyclase inhibitor), Rp-8-Br-PET-cGMPS (cGMP-dependent protein kinase inhibitor), SC560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethyl pyrazole, selective cyclooxygenase-1 inhibitor], and 4-aminopyridine (voltage-dependent K+ channel blocker) reduced AM-induced relaxation. On the other hand, 7-nitroindazole (selective neuronal nitric oxide synthase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), H89 (protein kinase A inhibitor), SQ22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, adenylate cyclase inhibitor], glibenclamide (selective blocker of ATP-sensitive K+ channels), and apamin (Ca2+-activated channel blocker) did not affect AM-induced relaxation. AM increased nitrate levels and 6-keto-PGF1α in rat CSM. The major new contribution of this research is that it demonstrated expression of AM and its receptor in rat CSM. Moreover, we provided evidence that AM-induced relaxation in this tissue is mediated by AM receptors by a mechanism that involves the nitric oxide-cGMP pathway, a vasodilator prostanoid, and the opening of voltage-dependent K+ channels.
Assuntos
Animais , Masculino , Adrenomedulina/farmacologia , Proteína Semelhante a Receptor de Calcitonina/análise , Músculo Liso/efeitos dos fármacos , Parassimpatolíticos/farmacologia , Pênis/efeitos dos fármacos , Vasodilatadores/farmacologia , /farmacologia , /análise , Adrenomedulina/genética , Adrenomedulina/metabolismo , Western Blotting , Proteína Semelhante a Receptor de Calcitonina/antagonistas & inibidores , Proteínas Quinases Dependentes de GMP Cíclico/antagonistas & inibidores , Inibidores de Ciclo-Oxigenase/farmacologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Imuno-Histoquímica , Indazóis/farmacologia , Relaxamento Muscular , Músculo Liso/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico/análise , Óxido Nítrico/análogos & derivados , Pênis/metabolismo , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , RNA Mensageiro/metabolismo , Proteína 1 Modificadora da Atividade de Receptores/genética , Proteína 1 Modificadora da Atividade de Receptores/metabolismo , /metabolismo , /genética , /metabolismo , Receptores de Peptídeo Relacionado com o Gene de Calcitonina/metabolismoRESUMO
The aim of the present study was to determine the mechanisms underlying the relaxant effect of adrenomedullin (AM) in rat cavernosal smooth muscle (CSM) and the expression of AM system components in this tissue. Functional assays using standard muscle bath procedures were performed in CSM isolated from male Wistar rats. Protein and mRNA levels of pre-pro-AM, calcitonin receptor-like receptor (CRLR), and Subtypes 1, 2 and 3 of the receptor activity-modifying protein (RAMP) family were assessed by Western immunoblotting and quantitative real-time polymerase chain reaction, respectively. Nitrate and 6-keto-prostaglandin F(1α) (6-keto-PGF(1α); a stable product of prostacyclin) levels were determined using commercially available kits. Protein and mRNA of AM, CRLR, and RAMP 1, -2, and -3 were detected in rat CSM. Immunohistochemical assays demonstrated that AM and CRLR were expressed in rat CSM. AM relaxed CSM strips in a concentration-dependent manner. AM(22-52), a selective antagonist for AM receptors, reduced the relaxation induced by AM. Conversely, CGRP(8-37), a selective antagonist for calcitonin gene-related peptide receptors, did not affect AM-induced relaxation. Preincubation of CSM strips with N(G)-nitro-L-arginine-methyl-ester (L-NAME, nitric oxide synthase inhibitor), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, quanylyl cyclase inhibitor), Rp-8-Br-PET-cGMPS (cGMP-dependent protein kinase inhibitor), SC560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethyl pyrazole, selective cyclooxygenase-1 inhibitor], and 4-aminopyridine (voltage-dependent K(+) channel blocker) reduced AM-induced relaxation. On the other hand, 7-nitroindazole (selective neuronal nitric oxide synthase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), H89 (protein kinase A inhibitor), SQ22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, adenylate cyclase inhibitor], glibenclamide (selective blocker of ATP-sensitive K(+) channels), and apamin (Ca(2+)-activated channel blocker) did not affect AM-induced relaxation. AM increased nitrate levels and 6-keto-PGF1α in rat CSM. The major new contribution of this research is that it demonstrated expression of AM and its receptor in rat CSM. Moreover, we provided evidence that AM-induced relaxation in this tissue is mediated by AM receptors by a mechanism that involves the nitric oxide-cGMP pathway, a vasodilator prostanoid, and the opening of voltage-dependent K(+) channels.
Assuntos
Adrenomedulina/farmacologia , Proteína Semelhante a Receptor de Calcitonina/análise , Músculo Liso/efeitos dos fármacos , Parassimpatolíticos/farmacologia , Pênis/efeitos dos fármacos , Vasodilatadores/farmacologia , 4-Aminopiridina/farmacologia , 6-Cetoprostaglandina F1 alfa/análise , Adrenomedulina/genética , Adrenomedulina/metabolismo , Animais , Western Blotting , Proteína Semelhante a Receptor de Calcitonina/antagonistas & inibidores , Proteínas Quinases Dependentes de GMP Cíclico/antagonistas & inibidores , Inibidores de Ciclo-Oxigenase/farmacologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Imuno-Histoquímica , Indazóis/farmacologia , Masculino , Relaxamento Muscular , Músculo Liso/metabolismo , Óxido Nítrico/análogos & derivados , Óxido Nítrico/análise , Óxido Nítrico Sintase/antagonistas & inibidores , Pênis/metabolismo , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , RNA Mensageiro/metabolismo , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Proteína 1 Modificadora da Atividade de Receptores/genética , Proteína 1 Modificadora da Atividade de Receptores/metabolismo , Proteína 2 Modificadora da Atividade de Receptores/genética , Proteína 2 Modificadora da Atividade de Receptores/metabolismo , Proteína 3 Modificadora da Atividade de Receptores/metabolismo , Receptores de Peptídeo Relacionado com o Gene de Calcitonina/metabolismoRESUMO
Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70â µM) for 24, 48, and 72â h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48â h of treatment with 50â µM KA, and 31% for cells submitted to 48â h of treatment with 70â µM KA. Similarly, in U87 cells treated with KA for 48â h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.
Assuntos
Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Diterpenos/farmacologia , Glioblastoma/tratamento farmacológico , Mikania/química , Caspase 3/efeitos dos fármacos , Caspase 8/efeitos dos fármacos , Linhagem Celular Tumoral , Diterpenos/isolamento & purificação , Proteína Ligante Fas , Citometria de Fluxo , Glioblastoma/enzimologia , Glioblastoma/patologia , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Fatores de TempoRESUMO
Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM) for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.
Assuntos
Humanos , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Diterpenos/farmacologia , Glioblastoma/tratamento farmacológico , Mikania/química , Linhagem Celular Tumoral , /efeitos dos fármacos , /efeitos dos fármacos , Diterpenos/isolamento & purificação , Proteína Ligante Fas , Citometria de Fluxo , Glioblastoma/enzimologia , Glioblastoma/patologia , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Fatores de TempoRESUMO
OBJECTIVE: To analyze the expression and distribution patterns of mature dendritic cells (mDCs) and immature DCs (imDCs) in radicular cysts (RCs), dentigerous cysts (DtCs), and keratocystic odontogenic tumors (KCOTs). MATERIALS AND METHODS: Forty-nine odontogenic cystic lesions (OCLs) (RCs, n = 20; DtCs, n = 15; KCOTs, n = 14) were assessed using the following markers: S100, CD1a and CD207 for imDCs; and CD83 for mDCs. RESULTS: Almost all cases were S100, CD1a, and CD207 positive, whereas 63% were CD83 positive. RCs presented greater number of immunostained cells, followed by DtCs, and KCOTs. The number of S100+ cells was greater than both CD1a+ and CD207+ cells (P < 0.001), which showed approximately similar amounts, followed by lower number of CD83+ cells (P < 0.001) in each OCL type. Different from S100+ cells, both CD1a+ and CD207+ cells on the epithelium (P < 0.05) and CD83+ cells on the capsule (P < 0.05) were preferentially observed. In RCs, significant correlation was found between the thickness epithelium with S100+ and CD1a+ cells, and between the degree of inflammation with CD83+ cells. CONCLUSIONS: Dendritic cell populations in OCLs can be phenotypically heterogeneous, and it could represent distinct lineages and/or functional stages. It is suggested that besides DC-mediated immune cell interactions, DC-mediated tissue differentiation and maintenance in OCLs should also be considered.