RESUMO
Two key enzymes like α-glucosidase and dipeptidyl peptidase-4 (DPP4) play an important role in the development of Type 2 diabetes (T2D), and inhibiting them is a common treatment strategy for T2D. In this study, an ultrafiltration- centrifugation approach was developed to isolate peptides from dark protein extract. Consequently, 6 peptides TAELLPR, CGKKFVR, AVPANLVDLNVPALLK, VVDLVFFAAAK, MSLYPR and QGQELLPSDFK were identified. The four peptides exhibited α-glucosidase inhibition activities with the IC50 values of 0.43 ± 0.03, 0.52 ± 0.09, 1.03 ± 0.13 and 0.04 ± 0.04 mg/mL, respectively, which were lower than the IC50 value of positive drug acarbose (1.51 ± 0.23 mg/mL); the last two peptides had DPP4 inhibition activities with the IC50 values of 1.35 ± 0.15 and 3.89 ± 0.22 mg/mL, respectively. This suggests that dark tea-derived peptides, for the first time, were found to be potential agents for α-glucosidase and DPP4 inhibition.
Assuntos
Inibidores da Dipeptidil Peptidase IV/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Peptídeos/farmacologia , Extratos Vegetais/farmacologia , Proteínas de Plantas/química , Chá/química , Dipeptidil Peptidase 4/metabolismo , Inibidores da Dipeptidil Peptidase IV/química , Inibidores da Dipeptidil Peptidase IV/isolamento & purificação , Relação Dose-Resposta a Droga , Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/isolamento & purificação , Humanos , Estrutura Molecular , Peptídeos/química , Peptídeos/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Relação Estrutura-Atividade , alfa-Glucosidases/metabolismoRESUMO
This study aims to elucidate the potential targets and molecular mechanisms underlying the anticancer effects of Red fermented rice extract using molecular simulation techniques. The inhibitory effects of different elution fractions of Red fermented rice extract on A549 and MCF-7 cell proliferation were evaluated through CCK-8 assays. Liquid chromatography-mass spectrometry (LC-MS) was employed to elucidate the structural information of active components, while molecular simulation techniques aided in identifying target proteins based on small molecule structures. Protein immunoblotting was utilized to investigate the mechanisms of action of relevant targets. The study found that the petroleum ether-ethyl acetate and ethyl acetate elution fractions of Red fermented rice extract significantly inhibited A549 and MCF-7 cell proliferation, with stronger effects observed on A549 cells. LC-MS structural analysis identified 25 small molecule structures. Molecular simulations successfully revealed interaction between active elution fractions of Red fermented rice extract and the cancer-related protein FGFR1. Further investigation into the phosphorylation of FGFR1 and its downstream pathway targets PI3K/AKT demonstrated that the active elution fractions exerted their anticancer activity by inhibiting the phosphorylation of FGFR1, PI3K, and AKT proteins. This comprehensive study, integrating CCK-8 assays, LC-MS, molecular simulation techniques, and protein immunoblotting, provides a deep understanding of the anticancer mechanisms of Red fermented rice extract, guiding its further development and clinical application.
Assuntos
Proliferação de Células , Fermentação , Oryza , Fosfatidilinositol 3-Quinases , Extratos Vegetais , Proteínas Proto-Oncogênicas c-akt , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos , Transdução de Sinais , Oryza/química , Humanos , Transdução de Sinais/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Células A549 , Células MCF-7 , Antineoplásicos Fitogênicos/farmacologia , Antineoplásicos Fitogênicos/isolamento & purificação , Fosforilação , Estrutura MolecularRESUMO
Spirulina platensis is a blue-green algae with potential anti-obesity effects. In this study, the anti-obesity effects of whole Spirulina platensis (WSP), Spirulina platensis protein (SPP) and Spirulina platensis protein hydrolysate (SPPH) were compared in high-fat diet fed mice, and the potential acting mechanism of SPPH was also investigated. Totally, SPPH exhibited good anti-obesity effects (reducing 39.8%±9.7% of body weight), lowering 23.8%±1.6% of serum glucose, decreasing 20.8%±1.4% of total cholesterol, while positive drug Simvastatin had the corresponding values: 8.3%±4.6%, 24.8%±1.9% and -2.1%±0.2%, respectively. Subsequently, PCR array was used to conduct gene expression analysis in brain and liver tissues of SPPH-treated mice, which displayed distinctly different expression pattern. The most markedly changed genes included: Acadm (-34.7 fold), Gcg (2.5 fold), Adra2b (2 fold) and Ghsr (2 fold) in brain; Retn (39 fold), Fabp4 (15.5 fold), Ppard (6 fold) and Slc27a1 (5.4 fold) in liver. Further network analysis demonstrated that the significantly expressed genes in brain and liver tissues were mapped into an interacting network, suggesting a modulatory effect on brain-liver axis, major pathways were involved in the axis: PPAR, adipocytokine, AMPK, non-alcoholic fatty liver disease and MAPK. This study showed that Spirulina platensis protein hydrolysate possessed anti-obesity effect in mice.