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1.
Sci Rep ; 11(1): 18869, 2021 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-34552185

RESUMO

The dynamic ability of adipocytes in adipose tissue to store lipid in response to changes in the nutritional input and inflammatory elicitors has a major impact on human health. Previously, we established laminarin-coated beads or LCB as an inflammatory elicitor for adipocytes. However, it was not clear whether LCB inhibits lipid accumulation in adipocytes. Here, we show that LCB acts in the early stage of adipogenesis through both interleukin-1 receptor-associated kinases (IRAK) and spleen tyrosine kinase (SYK) pathways, resulting in the activation of the AMP-activated protein kinase (AMPK) and nuclear factor-κB (NF-κB) complexes, which subsequently cause cell cycle arrest, downregulation of the key transcription factors and enzymes responsible for adipogenesis, inhibition of adipogenesis, and stimulation of an inflammatory response. While LCB could effectively block lipid accumulation during the early stage of adipogenesis, it could stimulate an inflammatory response at any stage of differentiation. Additionally, our results raise a possibility that toll-like receptor 2 (TLR2) and C-type lectin domain family 7 member A (CLEC7A/Dectin-1) might be potential ß-glucan receptors on the fat cells. Together, we present the mechanism of LCB, as fungal-like particles, that elicits an inflammatory response and inhibits adipogenesis at the early stage of differentiation.


Assuntos
Adipogenia/fisiologia , Glucanos/farmacologia , Células 3T3-L1 , Proteínas Quinases Ativadas por AMP/metabolismo , Adipócitos/metabolismo , Animais , Pontos de Checagem do Ciclo Celular , Glucanos/metabolismo , Inflamação , Camundongos , NF-kappa B/metabolismo , Fatores de Transcrição
2.
Sci Rep ; 10(1): 9437, 2020 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-32523023

RESUMO

Adipocytes from white-adipose tissue are known to produce inflammatory cytokines, which play a major role in energy balance and metabolism. While they can respond to pathogen-associated molecular pattern (PAMPs) such as lipopolysaccharide (LPS) from bacteria, it is not known whether adipocytes can be stimulated by fungal cells. Previously, adipocytes were shown to produce toll-like receptor 2 (TLR2), a ß-glucan receptor, suggesting that they could respond to ß-glucan on the fungal cell wall. In this study, we show that heat-killed yeast induce an inflammatory response in adipocytes. Using fungal-like particles, namely laminarin-coated beads (LCB), we find that these particles trigger the expression of many key inflammatory genes in dose- and time-dependent fashions in adipocytes. These results suggest that ß-glucan on the fungal cell wall is sufficient to elicit an inflammatory response in adipocytes. In addition, we show that both LCB and LCB-treated conditioned medium from RAW 264.7 murine macrophages (LCB-RM) induce the expression of those inflammatory genes through IKKß-IκBα proteins. Together, we conclude that the fungal-like particles and the conditioned medium elicit an inflammatory response in adipocytes through the canonical or classical NF-κB pathway.


Assuntos
Tecido Adiposo Branco/imunologia , Tecido Adiposo Branco/metabolismo , Glucanos/metabolismo , Células 3T3-L1 , Adipócitos/metabolismo , Tecido Adiposo Branco/fisiologia , Animais , Meios de Cultivo Condicionados/farmacologia , Citocinas/metabolismo , Glucanos/farmacologia , Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Camundongos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , beta-Glucanas/metabolismo
3.
Pharmacogn Mag ; 13(Suppl 4): S817-S821, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29491638

RESUMO

BACKGROUND: Obesity is one of the imperative dynamics in the incidence and intensification of type 2 diabetes mellitus (T2DM). Rhinacanthus nasutus leaf extracts are previously reported for their antidiabetic and antiobesity potential. OBJECTIVE: The present study was performed to evaluate glucose uptake stimulatory and antiadipogenic activities of a standardized rhinacanthins-rich extract (RRE) and its marker compounds namely rhinacanthin-C (RC), rhinacanthin-D (RD), and rhinacanthin-N (RN) in 3T3-L1 and L6 cells. MATERIALS AND METHODS: RRE was prepared by a green extraction process, and the marker compounds (RC, RD, and RN) were isolated from the RRE using a silica gel column chromatography. Glucose uptake stimulation in both 3T3-L1 and L6 cells was performed by quantification of residual glucose in the media using glucose oxidase kit. Antiadipogenic activity in 3T3-L1 adipocytes was performed by intracellular lipids quantification using oil red O dye. RESULTS: At the highest effective dose, RRE (20 µg/mL) exhibited satisfactory glucose uptake stimulatory effect in 3T3-L1 adipocytes that equivalent to RN (20 µg/mL) and the positive control insulin (0.58 µg/mL) but higher than RC (20 µg/mL) and RD (20 µg/mL). In addition, treatments of L6 myotubes showed that RRE (2.5 µg/mL) exhibited potent and equivalent glucose uptake stimulation (>80%) to RC (2.5 µg/mL) and the standard drugs, insulin (2.90 µg/mL) and metformin (219.5 µg/mL), but higher than RD (2.5 µg/mL) and RN (2.5 µg/mL). Furthermore, RRE (20 µg/mL) exhibited potent antiadipogenic effect in 3T3-L1 adipocytes, which equivalent to RC (20 µg/mL) but higher than RD (20 µg/mL) and RN (20 µg/mL). CONCLUSIONS: The undertaken study suggests that RRE could be used as an effective remedy in the treatment of obesity-associated T2DM. SUMMARY: Rhinacanthins-rich extract and its marker compounds showed potent glucose uptake stimulatory activity in 3T3-L1 adipocytes and L6 myotubesRhinacanthins-rich extract and rhinacanthin-C showed comparable antiadipogenic effect in 3T3-L1 adipocytesRRE could be used as an effective remedy in the treatment of obesity-associated T2DM. Abbreviations used: T2DM: Type-2 diabetes mellitus; RRE: Rhinacanthins-rich extract; RC: Rhinacanthin-C; RD: Rhinacanthin-D; RN: Rhinacanthin-N; α-MEM: α-Minimum essential medium; DMEM: Dulbecco's modified Eagle's medium; HS: Horse serum; FBS: Fetal bovine serum; BSA: Bovine serum albumin; IBMX: 3-isobutyl-1-methylxanthine; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; GO: Glucose oxidase; NMR: Nuclear magnetic resonance; HPLC: High-performance liquid chromatography.

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