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1.
Braz. j. biol ; 84: e254011, 2024. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1355886

RESUMO

Abstract Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P<0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.


Resumo A pecuária é uma parte fundamental da indústria agrícola no Paquistão e contribui com 11,53% do PIB nacional. Entre as espécies de gado, os búfalos são considerados o ouro negro do Paquistão. Sendo os maiores produtores de leite em todo o mundo, os búfalos Nili-Ravi são os mais famosos. Os búfalos são afetados por muitas doenças endêmicas, entre as quais a "septicemia hemorrágica" (SH). Este estudo busca verificar os efeitos da exposição experimental de P. multocida B:2 (oral) e seus imunógenos, ou seja, LPS (oral e intravenoso) e OMP (oral e subcutâneo), nos perfis hormonais reprodutivos em búfalos Nili-Ravi. Amostras de soro repetidas foram coletadas da veia jugular de animais experimentais por 21 dias (0, 2, 4, 8, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 e 504 horas). Os ensaios hormonais para determinar as concentrações séricas do hormônio liberador de gonadotrofina (GnRH), hormônio foliculoestimulante (FSH), hormônio luteinizante (LH), estrogênio (E2) e progesterona (P4) foram realizados usando kits comerciais Elisa (MyBioSource). O perfil hormonal de todos os grupos de tratamento das novilhas bubalinas apresentou variações significativas (P < 0,05) em relação ao grupo controle (G-1). Esses resultados indicam supressão no perfil hormonal reprodutivo de búfalos Nili-Ravi na exposição a P. multocida B:2 e seus imunógenos. Essa influência garante que a exposição à SH possa ser uma possível razão para o atraso da puberdade e o baixo desempenho reprodutivo em búfalos Nili-Ravi.


Assuntos
Animais , Feminino , Infecções por Pasteurella/veterinária , Reprodução , Hormônios Esteroides Gonadais/sangue , Búfalos , Progesterona , Bovinos , Lipopolissacarídeos , Hormônio Liberador de Gonadotropina , Pasteurella multocida
2.
Braz. J. Anesth. (Impr.) ; 73(4): 446-454, 2023. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1447616

RESUMO

Abstract Background Sepsis and septic shock still represent great challenges in critical care medicine. Sildenafil has been largely used in the treatment of pulmonary arterial hypertension, but its effects in sepsis are unknown. The aim of this study was to investigate the hypothesis that sildenafil can attenuate endotoxin-induced pulmonary hypertension in a porcine model of endotoxemia. Methods Twenty pigs were randomly assigned to Control group (n = 10), which received saline solution; or to Sildenafil group (n = 10), which received sildenafil orally (100 mg). After 30 minutes, both groups were submitted to endotoxemia with intravenous bacterial lipopolysaccharide endotoxin (LPS) infusion (4 µg.kg-1.h-1) for 180 minutes. We evaluated hemodynamic and oxygenation functions, and also lung histology and plasma cytokine (TNFα, IL-1β, IL6, and IL10) and troponin I response. Results Significant hemodynamic alterations were observed after 30 minutes of LPS continuous infusion, mainly in pulmonary arterial pressure (from Baseline 19 ± 2 mmHg to LPS30 52 ± 4 mmHg, p< 0.05). There was also a significant decrease in PaO2/FiO2 (from Baseline 411 ± 29 to LPS180 334 ± 49, p< 0.05). Pulmonary arterial pressure was significantly lower in the Sildenafil group (35 ± 4 mmHg at LPS30, p< 0.05). The Sildenafil group also presented lower values of systemic arterial pressure. Sildenafil maintained oxygenation with higher PaO2/FiO2 and lower oxygen extraction rate than Control group but had no effect on intrapulmonary shunt. All cytokines and troponin increased after LPS infusion in both groups similarly. Conclusion Sildenafil attenuated endotoxin-induced pulmonary hypertension preserving the correct heart function without improving lung lesions or inflammation.


Assuntos
Animais , Endotoxemia , Hipertensão Pulmonar/tratamento farmacológico , Suínos , Lipopolissacarídeos/farmacologia , Endotoxinas/farmacologia , Citrato de Sildenafila/farmacologia , Hemodinâmica , Hipertensão Pulmonar/induzido quimicamente
3.
São Paulo; s.n; 2023. 64 p.
Tese em Português | LILACS | ID: biblio-1527176

RESUMO

Introdução: A inflamação é um fator presente na fisiopatologia de doenças, e, o intestino, órgão susceptível a disfunções e à disbiose, que está em constante exposição a microrganismos, pode estar associado a este processo. Por outro lado, aminoácidos de cadeia ramificada (ACR) podem apresentar efeitos anti-inflamatórios em linhagem celular intestinal. Objetivo: Investigar os efeitos da suplementação de ACR, in vitro, na modulação da resposta inflamatória e do estresse oxidativo induzida por LPS em modelo de células intestinais Caco-2. Métodos: As culturas de células foram distribuídas em seis grupos, sendo: dois grupos controle - controle com meio Dulbecco's Modified Eagle's Medium sem ACR (CTL0) e com ACR (CTL) - e quatro grupos suplementados - grupo leucina (LEU), grupo isoleucina (ISO), grupo valina (VAL) e associação de ACR (LIV). Foi adotado um protocolo de pré-tratamento, com os ACR e o LPS. A viabilidade celular foi avaliada pelo ensaio de redução do MTT (brometo de [3-(4,5-dimetiltiazol-2yl 2,5-difenil tetrazolium]). A análise de proteínas (mTOR, AKT, AMPK, NF-kB, TAK-1 e JNK) foi feita por Western Blotting. A dosagem de citocina IL-8 no sobrenadante de cultura celular foi realizada por meio do kit Multiplex para imunoensaio. Foram avaliados componentes do sistema antioxidante relacionado à glutationa (GSH, GSSG e GPx) por meio de kit comercial. Os resultados foram expressos como média ± erro padrão. Resultados: No teste de viabilidade celular por MTT, verificou-se que os grupos tratados com ACR e LPS não apresentaram comprometimento da viabilidade celular em relação ao grupo CTL sem LPS. Os grupos CTL0, CTL, VAL e LIV, quando estimulados com LPS, apresentaram maior capacidade de síntese, in vitro, de IL-8, bem como maior fosforilação das proteínas JNK e NF-kB em relação aos seus respectivos grupos sem LPS. A suplementação. Todavia, as células estimuladas com LPS e suplementadas com leucina e isoleucina apresentaram capacidade de síntese, in vitro, de IL-8 e fosforilação das proteínas JNK e NF-kB que não diferiram significativamente das células suplementadas com esses aminoácidos e não estimuladas com LPS. A ausência de ACR (grupo CTL0) e a suplementação desses aminoácidos, in vitro, em células intestinais Caco-2, tratadas com ou sem LPS, não induziram alteração da atividade da enzima GPx e da razão intracelular GSH/GSSG. Conclusões: Os aminoácidos leucina e isoleucina apresentaram potencial efeito anti-inflamatório nas células Caco-2 por meio da modulação da fosforilação das proteínas JNK e NF-kB, cujo fato está associado à modulação da síntese, in vitro, de IL-8, a qual está envolvida na resposta inflamatória no intestino.


Introduction: Inflammation is a factor present in the pathophysiology of diseases, and the intestine, an organ susceptible to dysfunction and dysbiosis, which is constantly exposed to microorganisms, may be associated with this process. On the other hand, branched-chain amino acids (BCAAs) may have anti-inflammatory effects on intestinal cell lines. Objective: To investigate the effects of ACR supplementation, in vitro, on the modulation of inflammatory response and oxidative stress induced by LPS in a model of intestinal Caco-2 cells. Methods: The cell cultures were divided into six groups, as follows: two control groups - control with Dulbecco's Modified Eagle's Medium without ACR (CTL0) and with ACR (CTL) - and four supplemented groups - leucine group (LEU), isoleucine group (ISO), valine group (VAL) and ACR association (LIV). A pre-treatment protocol was adopted, with ACR and LPS. Cell viability was assessed by MTT reduction assay ([3-(4,5-dimethylthiazol-2yl 2,5-diphenyl tetrazolium] bromide)) Protein analysis (mTOR, AKT, AMPK, NF-kB, TAK -1 and JNK) was performed by Western Blotting. The dosage of cytokine IL-8 in the cell culture supernatant was performed using the Multiplex kit for immunoassay. Components of the antioxidant system related to glutathione (GSH, GSSG, and GPx) were evaluated by commercial kit. The results were expressed as mean ± standard error. Results: In the cell viability test by MTT, it was verified that the groups treated with ACR and LPS did not show impairment of cell viability compared to the CTL group without LPS. The CTL0, CTL, VAL, and LIV groups, when stimulated with LPS, showed a greater capacity for in vitro synthesis of IL-8, as well as greater phosphorylation of JNK and NF-kB proteins in relation to their respective groups without LPS. However, cells stimulated with LPS and supplemented with leucine and isoleucine showed in vitro synthesis capacity of IL-8 and phosphorylation of JNK and NF-kB proteins that did not differ significantly from cells supplemented with these amino acids and not stimulated with LPS. The absence of ACR (CTL0 group) and the supplementation of these amino acids, in vitro, in intestinal Caco-2 cells, treated with or without LPS, did not induce changes in the activity of the GPx enzyme and the intracellular GSH/GSSG ratio. Conclusions: The amino acids leucine and isoleucine had a potential anti-inflammatory effect on Caco-2 cells by modulating the phosphorylation of JNK and NF-kB proteins, which fact is associated with the modulation of the in vitro synthesis of IL-8, which is involved in the inflammatory response in the gut.


Assuntos
Lipopolissacarídeos , Células CACO-2 , Aminoácidos de Cadeia Ramificada , Inflamação , Mucosa Intestinal
4.
Braz. j. biol ; 83: e245202, 2023. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1285622

RESUMO

Abstract Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 μg/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 μg/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 μg/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.


Resumo Embora a própolis tenha sido relatada por ter atividade anti-inflamatória, seus efeitos no sistema complemento, uma parte do sistema imunológico inato, não foram muito estudados. Esta pesquisa foi conduzida para descobrir os efeitos da própolis da Indonésia nos níveis de expressão de C3, C1r/s, Bf, MBL e C6 em larvas de peixe-zebra induzidas por lipopolissacarídeo (LPS). Foram realizadas contagens de macrófagos que migram para o saco vitelino e histologia do fígado. As larvas foram divididas em quatro grupos: CON (cultivadas apenas em meio E3), LPS (cultivadas em meio contendo 0,5 μg/L de LPS), LPSIBU (cultivadas em meio contendo LPS e, em seguida, tratadas com 100 μg/L de ibuprofeno por 24 horas) e LPSPRO (cultivado em meio contendo LPS, e então imerso em própolis 14,000 μg/L por 24 horas). Os resultados mostraram que a expressão do gene do complemento em larvas dos grupos LPSIBU e LPSPRO foi geralmente menor que em larvas do grupo LPS. O número de migrações de macrófagos para a gema no grupo LPSPRO também foi menor que no grupo LPS. A estrutura histológica do fígado em todos os grupos foi considerada normal. Este estudo mostra que a própolis indonésia tem potencial para ser utilizada como alternativa na substituição dos AINEs (anti-inflamatórios não esteroides).


Assuntos
Animais , Própole/farmacologia , Peixe-Zebra/genética , Regulação para Baixo , Lipopolissacarídeos/farmacologia , Indonésia , Larva/genética
5.
Biol. Res ; 55: 22-22, 2022. ilus, graf
Artigo em Inglês | LILACS | ID: biblio-1383923

RESUMO

BACKGROUND: Retinal neurodegeneration is induced by a variety of environmental insults and stresses, but the exact mechanisms are unclear. In the present study, we explored the involvement of cytosolic mitochondrial DNA (mtDNA), resulting in the cGAS-STING dependent inflammatory response and apoptosis in retinal damage in vivo. METHODS: Retinal injury was induced with white light or intravitreal injection of lipopolysaccharide (LPS). After light-or LPS-induced injury, the amount of cytosolic mtDNA in the retina was detected by PCR. The mtDNA was isolated and used to transfect retinas in vivo. WB and real-time PCR were used to evaluate the activation of cGAS-STING path-way and the levels of apoptosis-associated protein at different times after mtDNA injection. Retinal cell apoptosis rate was detected by TUNEL staining. Full-field electroretinography (ERG) was used to assess the retinal function. RESULTS: Light injury and the intravitreal injection of LPS both caused the leakage of mtDNA into the cytoplasm in retinal tissue. After the transfection of mtDNA in vivo, the levels of cGAS, STING, and IFN-ß mRNAs and the protein levels of STING, phosph-TBK1, phospho-IRF3, and IFN-ß were upregulated. mtDNA injection also induced the activation of caspase 3 and caspase 9. BAX and BAK were increased at both the mRNA and protein levels. The release of cytochrome c from the mitochondria to the cytosol was increased after mtDNA injection. The wave amplitudes on ERG decreased and retinal cell apoptosis was detected after mtDNA injection. CONCLUSIONS: Cytosolic mtDNA triggers an inflammatory response. It also promotes apoptosis and the dysfunction of the retina.


Assuntos
Animais , Ratos , DNA Mitocondrial/genética , Lipopolissacarídeos , Injeções Intravítreas , Proteínas de Membrana/metabolismo , Mitocôndrias , Nucleotidiltransferases/genética , Nucleotidiltransferases/metabolismo
6.
Braz. J. Pharm. Sci. (Online) ; 58: e21530, 2022. graf
Artigo em Inglês | LILACS | ID: biblio-1420486

RESUMO

Abstract Therapeutically, piracetam has been used for decades as a cognitive enhancer for memory- related neuronal disorders. The present study aimed to investigate the neuroprotective potential of piracetam on lipopolysaccharides (LPS)-induced neuronal deficit using both in-vitro and in-vivo experimental models. For the in-vitro analysis, EOC-20 murine microglial cells were induced with a neuronal toxicity of 100 µg/ml of LPS, and the formation of intracellular reactive oxygen species (ROS) and nitric oxide (NO) productions were determined. For in-vivo neuroprotective analysis, groups of mice were treated orally with two doses of piracetam (200 and 400 mg/kg) for 30 days. Neuronal toxicity was induced by four intraperitoneal injections of LPS (250 µg/kg/day). The malondialdehyde (MDA) level was measured for oxidative stress, and catalase reduced glutathione (GSH), glutathione reductase (GRD), and superoxide dismutase (SOD) levels were determined as the antioxidant parameters. The result of the cell viability study was that pre-treatment with piracetam significantly protected the LPS-induced cell loss, and attenuated the ROS generation and NO production in LPS-induced EOC-20 cells. Moreover, the treatment of piracetam significantly reduced the MDA levels and improved catalase, GSH, GRD, and SOD activities in LPS-induced mice brains. The overall results from this study supported the neuroprotective effects of piracetam against LPS-induced neuronal toxicity.


Assuntos
Animais , Masculino , Camundongos , Piracetam/análise , Lipopolissacarídeos/farmacologia , Neuroproteção/efeitos dos fármacos , Estresse Oxidativo , Cérebro/anormalidades , Doenças Neuroinflamatórias/induzido quimicamente , Antioxidantes/efeitos adversos
7.
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-1143396

RESUMO

ABSTRACT Objective: To determine the impact of distinctive instrumentation systems of the root canals on the endotoxin lessening through the root canals. Material and Methods: From the electronic databases, MEDLINE, PubMed, Cochrane Library, Embase, ISI, Google Scholar have been used to perform a systematic literature review between 2015 and 2020. Therefore, a software program (Endnote X9) has been utilized for managing electronic titles. Searches were performed with keywords, "root canal," "instrumentation," "endotoxin," "root canal preparation," "biofilm" "endodontics," and "lipopolysaccharide." This systematic review has been conducted on the basis of the key consideration of the PRISMA Statement-Preferred Reporting Items for the Systematic Review and Meta-analysis. Results: Hence, 163 potentially important abstracts and research topics have been discovered by electronic searches and three studies (3 RCTs) have been included. According to the outputs, any statistically significant differences have been not found between the rotary files and reciprocation (SMD 0.51, 95% CI [0.11, 0.90], p=0.011) (I2 = 49.5%; p=0.138). Conclusion: Analyses indicated that instrumentation methods decreased the content of endotoxin from the root canals.


Assuntos
Lipopolissacarídeos , Metanálise como Assunto , Preparo de Canal Radicular , Cavidade Pulpar , Endodontia , Irã (Geográfico)
8.
Braz. j. med. biol. res ; 54(11): e11215, 2021. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1285662

RESUMO

This study investigated the acute blockade of endogenous melatonin (MLT) using Luzindole with or without systemic lipopolysaccharide (LPS) challenge and evaluated changes in inflammatory and oxidative stress markers in the mouse jejunum. Luzindole is an MT1/MT2 MLT receptor antagonist. Both receptors occur in the small intestine. Swiss mice were treated with either saline (0.35 mg/kg, ip), Luzindole (0.35 mg/kg, ip), LPS (1.25 mg/kg, ip), or Luzindole+LPS (0.35 and 1.25 mg/kg, ip, respectively). Jejunum samples were evaluated regarding intestinal morphometry, histopathological crypt scoring, and PAS-positive villus goblet cell counting. Inflammatory Iba-1, interleukin (IL)-1β, tumor necrosis factor (TNF)-α, nuclear factor (NF)-kB, myeloperoxidase (MPO), and oxidative stress (NP-SHs, catalase, MDA, nitrate/nitrite) markers were assessed. Mice treated with Luzindole, LPS, and Luzindole+LPS showed villus height shortening. Crypt damage was worse in the LPS group. Luzindole, LPS, and Luzindole+LPS reduced the PAS-goblet cell labeling and increased Iba-1-immunolabelled cells compared to the saline group. Immunoblotting for IL-1β, TNF-α, and NF-kB was greater in the Luzindole group. The LPS-challenged group showed higher MPO activity than the saline and Luzindole groups. Catalase was reduced in the Luzindole and Luzindole+LPS groups compared to saline. The Luzindole group showed an increase in NP-SHs, an effect related to compensatory GSH activity. The acute blockade of endogenous MLT with Luzindole induced early changes in inflammatory markers with altered intestinal morphology. The other non-detectable deleterious effects of Luzindole may be balanced by the unopposed direct action of MLT in immune cells bypassing the MT1/MT2 receptors.


Assuntos
Animais , Ratos , Lipopolissacarídeos , Melatonina , Triptaminas , Inflamação/induzido quimicamente , Jejuno
9.
Braz. j. med. biol. res ; 54(7): e10603, 2021. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1249320

RESUMO

Neonatal sepsis is an inflammatory system syndrome and a main cause of neonatal mortality. However, there is a lack of ideal biomarkers for early neonatal sepsis diagnosis. The aim of this study was to evaluate the clinical significance of miR-141 in sepsis in neonates, and explore the regulatory effects of miR-141 on inflammation in monocytes. This study used qRT-PCR to calculate the expression of miR-141 in the serum of septic neonates. The diagnostic values of procalcitonin (PCT) and serum miR-141 were evaluated by receiver operating characteristic (ROC) curves. The relationship between miR-141 and TLR4 was determined using luciferase reporter assay. An inflammation model was established using monocytes with lipopolysaccharide (LPS) treatment. ELISA assay was used to analyze the levels of pro-inflammatory cytokines. The expression of miR-141 in neonatal sepsis was significantly lower than healthy controls. ROC curves showed that miR-141 had diagnostic accuracy. LPS stimulation in monocytes led to a decrease in the expression of miR-141. A luciferase reporter assay proved that miR-141 targeted TLR4, and a negative correlation of miR-141 with TLR4 was found in septic neonates. ELISA results demonstrated that the overexpression of miR-141 inhibited LPS-induced inflammation in monocytes. In conclusion, serum decreased miR-141 expression served as a candidate diagnostic biomarker of neonatal sepsis. TLR4 is a target gene of miR-141, which may mediate the inhibitory effects of miR-141 overexpression on LPS-induced inflammation in monocytes. Therefore, miR-141 is expected to be a potential diagnostic biomarker and a therapeutic target in neonatal sepsis.


Assuntos
Humanos , Recém-Nascido , Sepse , MicroRNAs , Sepse Neonatal , Monócitos , Lipopolissacarídeos , Receptor 4 Toll-Like
10.
Arq. bras. med. vet. zootec. (Online) ; 72(6): 2397-2401, Nov.-Dec. 2020. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1142296

RESUMO

O presente estudo avaliou o efeito da suplementação com OmniGen-AF® na proliferação de linfócitos e títulos de anticorpos após vacinação em bovinos leiteiros. Amostras de sangue periférico foram coletadas de 32 vacas leiteiras para quantificação dos títulos de anticorpos anti-Leptospira, e amostras de sangue periférico de 16 vacas leiteiras foram também coletadas para avaliação da proliferação de linfócitos. Observou-se que a suplementação com OmniGen-AF® aumentou a proliferação basal de linfócitos (sem estímulos) 21 dias após a vacinação (P=0,03), apesar de reduzir a proliferação de linfócitos B quando estimulada com Leptospira borgpetersenii serovar Hardjo inativada pelo calor (P=0,03). Ademais, nenhum efeito da suplementação sobre a proliferação de linfócitos no momento imediatamente anterior à vacinação e nos títulos de anticorpos anti-Leptospira foi encontrado. Além disso, a proliferação de linfócitos estimulada com lipopolissacarídeos foi maior em vacas multíparas que em primíparas 21 dias após a vacinação (P=0,03). Desse modo, o presente estudo demonstra que a suplementação com OmniGen-AF® não afetou de forma robusta a proliferação de linfócitos e os títulos de anticorpos anti-Leptospira após vacinação em vacas leiteiras sadias.(AU)


Assuntos
Animais , Feminino , Bovinos , Vacinas Combinadas/análise , Suplementos Nutricionais/análise , Fatores Imunológicos/administração & dosagem , Linfocitose/veterinária , Lipopolissacarídeos , Leptospira/imunologia
11.
J. bras. nefrol ; 42(3): 300-306, July-Sept. 2020. tab, graf
Artigo em Inglês, Português | LILACS | ID: biblio-1134857

RESUMO

ABSTRACT Introduction: Vascular calcification is a common complication of chronic kidney disease. Osteoblast differentiation factor (Cbfa1) is present in histologic sections of arteries from patients with end-stage renal disease. Vascular smooth muscle cells (VSMC) can dedifferentiate to osteoblast-like cells, possibly by up-regulation of Cbfa1. There is evidence that the production of nitric oxide (NO) may have an important role in the regulation of osteoblast metabolism. The aim of this study is to evaluate whether increased NO/iNOS expression causes an increase in cbfa1 expression in VSMC. Methods: VSMC were obtained from renal artery of Wistar male rats, treated for 72 hours with lipopolysaccharide (LPS), ß-glycerophosphate (BGF), a donor of phosphate and aminoguanidine (AG), an inhibitor of iNOS, in the following groups: CTL (control), LPS, BGF, LPS + BGF, and LPS + AG. NO synthesis was determined by chemiluminescence. Cbfa1 and iNOS mRNA expressions were analyzed by RT-PCR, Cbfa1 protein expression by immunohistochemistry and cellular viability by acridine orange. Results: Cbfa1 and iNOS mRNA expressions were higher in LPS and LPS+ BGF vs CTL (p < 0.05), and they were lower in LPS+AG vs LPS (p < 0.05). The Cbfa1 in the groups LPS and LPS+BGF also resulted in a higher value compared to CTL (p < 0.05), and in LPS+AG it was lower compared to LPS (p < 0.05). NO was higher in LPS and LPS+BGF compared to CTL group (p < 0.05) and lower in LPS + AG compared to LPS group (p < 0.05). Cellular viability showed no statistical difference among groups. Conclusion: This study showed that increased NO/iNOS expression causes an increase in cbfa1 expression in VSMC.


RESUMO Introdução: A calcificação vascular é uma complicação comum da doença renal crônica. O fator de diferenciação osteoblástica (Cbfa1) está presente em cortes histológicos das artérias de pacientes com doença renal em estágio terminal. As células do músculo liso vascular (CMLV) podem desdiferenciar para células do tipo osteoblastos, possivelmente pela regulação positiva da Cbfa1. Há evidências de que a produção de óxido nítrico (NO) pode ter um papel importante na regulação do metabolismo dos osteoblastos. O objetivo deste estudo é avaliar se o aumento da expressão de NO/iNOS causa um aumento na expressão de cbfa1 nas CMLV. Métodos: As CMLV foram obtidas da artéria renal de ratos machos Wistar, tratados por 72 horas com lipopolissacarídeo (LPS), ß-glicerofosfato (BGF), um doador de fosfato e aminoguanidina (AG), um inibidor da iNOS, nos seguintes grupos: CTL (controle), LPS, BGF, LPS + BGF e LPS + AG. A síntese de NO foi determinada por quimioluminescência. As expressões de mRNA de Cbfa1 e iNOS foram analisadas por RT-PCR, a expressão da proteína Cbfa1 por imunohistoquímica e viabilidade celular por laranja de acridina. Resultados: As expressões de mRNA de Cbfa1 e iNOS foram maiores em LPS e LPS + BGF v.s. CTL (p < 0,05) e menores em LPS + AG v.s. LPS (p <0,05). O Cbfa1 nos grupos LPS e LPS + BGF também resultou em um valor maior em comparação ao CTL (p < 0,05), e no LPS + AG foi menor em comparação ao LPS (p < 0,05). NO foi maior no LPS e LPS + BGF em comparação ao grupo CTL (p < 0,05) e menor no LPS + AG em comparação ao grupo LPS (p < 0,05). A viabilidade celular não mostrou diferença estatística entre os grupos. Conclusão: Este estudo mostrou que o aumento da expressão de NO/iNOS causa um aumento na expressão de cbfa1 nas CMLV.


Assuntos
Humanos , Animais , Masculino , Ratos , Músculo Liso Vascular , Óxido Nítrico , Artéria Renal , Lipopolissacarídeos , Ratos Wistar , Subunidade alfa 1 de Fator de Ligação ao Core
12.
Acta cir. bras ; 35(11): e351102, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1141938

RESUMO

Abstract Purpose: In this experimental study, activated protein C (APC), which has anticoagulant, antithrombotic, profibrinolytic, anti-inflammatory and antiapoptotic properties, was used to prevent coagulopathy in a disseminated intravascular coagulation (DIC) model formatted with lipopolysaccharide (LPS) infusion. Methods: Twenty-five Wistar albino rats weighting 280 - 320 g each were used. They were randomly divided into three groups: sham, control and study groups. To sham group (n = 5), only normal saline was infused. To control (n = 10) and study groups (n = 10), 30 mg/kg LPS was infused for 4 h from femoral vein. After LPS infusion, 100 µg/kg recombinant APC was given during 4 h in study group. Eight hours later, blood samples were taken from abdominal aorta and the animals sacrificed. From these samples, platelet, prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen and D-dimer levels were studied. Results: Platelet counts and fibrinogen levels were significantly lower in control and study groups than sham group (p < 0.05). The PT, aPTT and D-dimer levels were significantly higher in control and study groups than in sham group (p < 0.05). When comparing control and study groups, platelet counts were not statistically different (p = 0.36). However, the difference of the fibrinogen levels was significant between these groups (p = 0.0001). While PT and aPTT were longer in the study group compared to the control group (p < 0.05), D-dimer levels were lower in the study group than in control (p = 0.0001). Conclusion: Use of APC can prevent hypercoagulation and consumption coagulopathy in the DIC as a result of correcting hematological parameters other than prolongation of coagulation time.


Assuntos
Animais , Coagulação Intravascular Disseminada/tratamento farmacológico , Proteína C , Lipopolissacarídeos , Ratos Wistar , Anticoagulantes
13.
Braz. arch. biol. technol ; 63: e20180612, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1132197

RESUMO

Abstract The present study aimed to evaluate the anti-inflammatory potential of a Lycium barbarum (L. barbarum) fruit extract in Wistar rats submitted to a palatable diet presenting systemic inflammation induced by lipopolysaccharides (LPS). Forty-two Wistar female rats (Rattus Novergicus) were used with 60 days old. The animals were feed for 60 days and divided in six groups (n=7): standard diet+water; standard diet+L. barbarum; palatable diet+water; palatable diet+L. barbarum; standard diet+water+LPS; standard diet+L. barbarum+LPS. A significant difference was shown between the analyzed groups concerning C-reactive protein, with the standard diet+water+LPS group presenting the highest inflammatory response in comparison to the other groups. Decreased inflammatory response was observed in the group administered a palatable diet along with the fruit extract when compared to the group that received only a palatable diet. Significant decrease in glutamic-oxaloacetic transaminase activity was observed in the standard diet+L. barbarum+LPS group compared to the standard diet+water group, as well as in the palatable diet+L. barbarum group compared to the palatable diet+water group. A significant increase in creatinine in the standard diet+water+LPS group was observed in according to the L. barbarum administration groups. The gene expression of the inflammatory markers genes in the liver showed a significant increase in TNF-α and IL-6 genes in the group treated with standard diet+L. barbarum+LPS when compared to the standard diet+LPS group. Thus, the administered L. barbarum extract displays the potential to reduce inflammatory responses induced by LPS and a palatable diet.


Assuntos
Animais , Feminino , Ratos , Lycium , Inflamação/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Extratos Vegetais , Lipopolissacarídeos/efeitos adversos , Ratos Wistar , Alanina Transaminase , Modelos Animais de Doenças , Inflamação/microbiologia
14.
Clinics ; 75: e1665, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1133413

RESUMO

OBJECTIVES: This study intended to explore the effect of T regulatory cells (Tregs) in the perinatal liver against LPS-induced inflammation in a preterm birth mouse model. Moreover, the role of adoptive Tregs on the inflammatory response induced by LPS was also studied. METHODS: Female BALB/C mice were injected intraperitoneally (IP) with LPS dissolved in normal saline solution at a dose of 50 µg/kg. Spleens from pregnant mice were used to obtain Tregs. The expression of Forkhead family transcription factor-3 (Foxp3), Interleukin-6 (IL-6), Toll-like receptor-4 (TLR-4), and Heme oxygenase-1 (HO-1) were assessed from fetal liver tissues by polymerase chain reaction and western blotting. RESULTS: LPS administered to mice induced an inflammatory response in the perinatal liver, and this inflammatory response was negatively regulated by Tregs in the experimental group. Maternal-fetal tolerance was maintained by Tregs. Transmission of Tregs was estimated in different experimental groups based on the mRNA expression of TLR-4, IL-6, HO-1, and Foxp3. CONCLUSIONS: After analysis of the experimental data, it was determined that Tregs exhibited regulatory potential against LPS-induced inflammatory response. Further, it was concluded that the transmission of Tregs improved the mother's immune tolerance against LPS-induced inflammation in the fetal liver.


Assuntos
Animais , Feminino , Gravidez , Camundongos , Lipopolissacarídeos/toxicidade , Nascimento Prematuro , Linfócitos T Reguladores , Fatores de Transcrição Forkhead , Inflamação/induzido quimicamente , Fígado , Camundongos Endogâmicos BALB C
15.
J. appl. oral sci ; 28: e20190699, 2020. graf
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-1134770

RESUMO

Abstract Purpose To evaluate the kinetics of apical periodontitis development in vivo , induced either by contamination of the root canals by microorganisms from the oral cavity or by inoculation of bacterial lipopolysaccharide (LPS) and the regulation of major enzymes and receptors involved in the arachidonic acid metabolism. Methodology Apical periodontitis was induced in C57BL6 mice (n=96), by root canal exposure to oral cavity (n=48 teeth) or inoculation of LPS (10 µL of a suspension of 0.1 µg/µL) from E. coli into the root canals (n= 48 teeth). Healthy teeth were used as control (n=48 teeth). After 7, 14, 21 and 28 days the animals were euthanized and tissues removed for histopathological and qRT-PCR analyses. Histological analysis data were analyzed using two-way ANOVA followed by Sidak's test, and qRT-PCR data using two-way ANOVA followed by Tukey's test (α=0.05). Results Contamination by microorganisms led to the development of apical periodontitis, characterized by the recruitment of inflammatory cells and bone tissue resorption, whereas inoculation of LPS induced inflammatory cells recruitment without bone resorption. Both stimuli induced mRNA expression for cyclooxygenase-2 and 5-lipoxygenase enzymes. Expression of prostaglandin E 2 and leukotriene B 4 cell surface receptors were more stimulated by LPS. Regarding nuclear peroxisome proliferator-activated receptors (PPAR), oral contamination induced the synthesis of mRNA for PPARδ, differently from inoculation of LPS, that induced PPARα and PPARγ expression. Conclusions Contamination of the root canals by microorganisms from oral cavity induced the development of apical periodontitis differently than by inoculation with LPS, characterized by less bone loss than the first model. Regardless of the model used, it was found a local increase in the synthesis of mRNA for the enzymes 5-lipoxygenase and cyclooxygenase-2 of the arachidonic acid metabolism, as well as in the surface and nuclear receptors for the lipid mediators prostaglandin E2 and leukotriene B4.


Assuntos
Animais , Masculino , Periodontite Periapical/microbiologia , Dinoprostona/metabolismo , Lipopolissacarídeos/metabolismo , Leucotrieno B4/metabolismo , Cavidade Pulpar/microbiologia , Periodontite Periapical/metabolismo , Periodontite Periapical/patologia , Fatores de Tempo , Reabsorção Óssea/metabolismo , Reabsorção Óssea/microbiologia , Araquidonato 5-Lipoxigenase/análise , Araquidonato 5-Lipoxigenase/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Dinoprostona/análise , Distribuição Aleatória , Expressão Gênica , Leucotrieno B4/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Cavidade Pulpar/metabolismo , Cavidade Pulpar/patologia , Ciclo-Oxigenase 2/análise , Ciclo-Oxigenase 2/metabolismo , Camundongos Endogâmicos C57BL
16.
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-1135491

RESUMO

Abstract Objective: To show the cytotoxicity of Porphyromonas gingivalis lipopolysaccharide (LPS) on human umbilical cord mesenchymal stem cells (HUCMSCs) to better understand the characteristics for its application in regenerative procedures under periodontopathogen LPS influence. Material and Methods: Ultrapure Porphyromonas gingivalis LPS was used in this study. This research used a frozen stock HUCMSCs, previously confirmed by flow cytometry. The biological characteristics, such as cell morphology, proliferation, and protein expression, were screened. To check the cytotoxicity, HUCMSCs were cultured and divided into two groups, the control group and LPS group with various concentrations from 25 to 0.39 µg/mL. MTT assay was done and the cells were observed and counted. The significance level was set at 5%. Results: The percentage of living HUCMSCs on LPS group were not significantly different among concentrations (p>0.05) from 25 to 0.39 µg/mL, even though there were slight mean decrease between groups, but they were not significant. The duration of 24 hours of exposure of LPS does not significantly lower HUCMSCs viability. Conclusion: LPS does not affect the viability of HUCMSCs. The lower the concentration of LPS, the higher the viability of HUCMSCs.


Assuntos
Humanos , Cordão Umbilical , Lipopolissacarídeos , Porphyromonas gingivalis , Citotoxicidade Imunológica/imunologia , Células-Tronco Mesenquimais , Análise de Variância , Citometria de Fluxo , Indonésia/epidemiologia
17.
J. appl. oral sci ; 28: e20190519, 2020. tab, graf
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-1101254

RESUMO

Abstract Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 μM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 μM (p<0.05) and 250 μM (p<0.01). The POH increased ROS production at both 10 μM and 100 μM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 μM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 μM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.


Assuntos
Animais , Camundongos , Fusobacterium nucleatum/efeitos dos fármacos , Espécies Reativas de Oxigênio/análise , Porphyromonas/efeitos dos fármacos , Monoterpenos/farmacologia , Macrófagos/efeitos dos fármacos , Antibacterianos/farmacologia , Arginase/análise , Fatores de Tempo , Produtos Biológicos/farmacologia , Testes de Sensibilidade Microbiana , Expressão Gênica , Lipopolissacarídeos/farmacologia , Reprodutibilidade dos Testes , Fator de Necrose Tumoral alfa/análise , Fusobacterium nucleatum/crescimento & desenvolvimento , Espécies Reativas de Oxigênio/metabolismo , Porphyromonas/crescimento & desenvolvimento , Proliferação de Células/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Citometria de Fluxo , Células RAW 264.7 , Macrófagos/metabolismo
18.
Braz. j. med. biol. res ; 53(12): e10109, 2020. graf
Artigo em Inglês | LILACS, Coleciona SUS | ID: biblio-1132504

RESUMO

Psoriasis is a chronic inflammatory skin disorder in humans, and the inflammatory reaction plays an important role in development and onset of psoriasis. 4'-O-β-D-glucosyl-5-O-methylvisamminol (4GMV) is one of the major active chromones isolated from Saposhnikoviae divaricata (Turcz.) Schischk, which has been reported to exhibit excellent anti-inflammatory activities. However, the possible therapeutic effect on psoriasis and underlying mechanism has not been reported. Thus, the aim of this study was to investigate the protective effect of 4GMV on the imiquimod (IMQ)-induced psoriasis-like lesions in BALB/c mice and the anti-inflammatory effect on the lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages. The results demonstrated that 4GMV decreased IMQ-induced keratinocyte proliferation and inflammatory cell infiltration. Moreover, 4GMV treatment significantly inhibited the production of NO, PEG 2, and cytokines such as interleukin (IL)-1β, IL-6, interferon (IFN)-γ, and IL-22 in LPS-stimulated RAW264.7 macrophages. 4GMV also suppressed the LPS-upregulated protein expressions of iNOS and COX-2 in a dose-dependent manner. Furthermore, qRT-PCR analysis showed that 4GMV down-regulated the mRNA level of IL-1β and IL-6 expression. Further studies by western blot indicated that 4GMV inhibited the activation of upstream mediator NF-κB by suppressing the expression of TLR4 and the phosphorylation of IκBα and p65. The phosphorylation of JNK, p38, and ERK were also markedly reversed by 4GMV in LPS-treated RAW264.7 macrophages. Taken together, these results demonstrated that 4GMV showed a protective effect in IMQ-induced psoriasis-like mice and inhibited inflammation through the NF-κB and MAPK signaling pathways, indicating that 4GMV might be a potential therapeutic drug for psoriasis.


Assuntos
Animais , Coelhos , Psoríase/induzido quimicamente , Psoríase/tratamento farmacológico , Dermatite , Lipopolissacarídeos , Citocinas , NF-kappa B , Cromonas , Sistema de Sinalização das MAP Quinases , Imiquimode , Glucosídeos , Inflamação , Camundongos Endogâmicos BALB C
19.
Acta cir. bras ; 35(2): e202000204, 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1100885

RESUMO

Abstract Purpose To investigate the effect of growth arrest-specific protein 6 (Gas6) on acute liver injury in mice and related mechanisms. Methods Thirty C57BL/6 (6-8 weeks old) mice were randomly divided into control, LPS/D-GalN, and LPS/D-GalN+Gas6 groups (10 mice in each group). The LPS/D-GalN group was intraperitoneally administered with LPS (0.25 mg/Kg) and D-GalN (400 mg/Kg) for 5h. The LPS/D-GalN+Gas6 group was intraperitoneally administered with rmGas6 one hour before intraperitoneal application of LPS/D-GalN. All subjects were sacrificed at 5 h for blood and tissue analysis. The expression of protein and mRNA was assessed by western blotting and RT-PCR, respectively. Results Compared with the control group, AST, ALT, IL-1β, TNF-α, IL-6 IL-10, MPO activity were increased in the LPS/D-GalN group. However, they were significantly inhibited by Gas6. Gas6 markedly suppressed the expression of apoptosis-related protein induced by LPS/D-GalN. Moreover, Gas6 attenuated the activation of the NF-κB signaling pathway in acute liver injury induced by LPS/D-GalN. Conclusions Gas6 alleviates acute liver injury in mice through regulating NF-κB signaling pathways. Gas6 can be a potential therapeutic agent in treating LPS/D-GalN-induced acute liver injury in the future.


Assuntos
Animais , Masculino , Camundongos , Lipopolissacarídeos/efeitos adversos , Apoptose/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Fígado/efeitos dos fármacos , Anti-Inflamatórios/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , Avaliação Pré-Clínica de Medicamentos , Anti-Inflamatórios/uso terapêutico
20.
Braz. oral res. (Online) ; 34: e013, 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1089379

RESUMO

Abstract This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFβ-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 μg/ml and treated with 15d-PGJ2 1 and 2 μg/ml, and ScLL 2 and 5 μg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFβ-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 μg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFβ-1 release (p > 0.05). Results indicated that ScLL 5 μg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFβ-1 release.


Assuntos
Humanos , Prostaglandina D2/análogos & derivados , Lipopolissacarídeos/farmacologia , Interleucina-6/metabolismo , Lectinas de Plantas/farmacologia , Fator de Crescimento Transformador beta1/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Valores de Referência , Fatores de Tempo , Ensaio de Imunoadsorção Enzimática , Prostaglandina D2/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Análise de Variância , Estatísticas não Paramétricas , Fator de Crescimento Transformador beta1/efeitos dos fármacos , Gengiva/citologia
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