RESUMEN
BACKGROUND: Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metabolites toward CRC cells is not clear. Therefore, the aim of this study was to assess the inhibitory mechanism of cell-free supernatants (CFSs) of L. rhamnosus GG, L. casei M3, and L. plantarum YYC-3 on metastasis of CRC cells. RESULTS: YYC-3 CFS showed the highest inhibitory effect on CRC cell growth, invasion and migration, and inhibited MMP2, MMP9, and VEGFA gene and protein expression, and protein secretion. Furthermore, it suppressed the activities of MMPs by gelatin zymography. Moreover, the effective compounds in these CFSs were analyzed by Q Exactive Focus liquid chromatography-mass spectrometry. CONCLUSIONS: Our results showed that metabolite secretions of YYC-3 may inhibited cell metastasis by downregulating the VEGF/MMPs signaling pathway. These data suggest that treatment of CRC cells with metabolites from L. plantarum YYC-3 may reduce colon cancer metastasis.
Asunto(s)
Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/microbiología , Lactobacillus/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Células CACO-2 , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica , Células HT29 , Humanos , Lacticaseibacillus casei/metabolismo , Lactobacillus plantarum/metabolismo , Lacticaseibacillus rhamnosus/metabolismo , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Metástasis de la Neoplasia/tratamiento farmacológico , Transducción de Señal/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
Dipeptidyl peptidase-IV (DPP-IV) is an enzyme that break down the antidiabetic hormone glucagon-like peptide-1. Therefore, inhibition of DPP-IV could be an effective strategy to treat Type 2 diabetes (T2D). The α-lactalbumin-rich whey protein concentrate was hydrolyzed by trypsin, and the hydrolysates were then fractionated at a semi-preparative scale using a Superdex Gel filtration Chromatography. The peptides were analyzed by using HPLC coupled with tandem mass spectrometry (RP-HPLC-MS/MS), and their Dipeptidyl peptidase-IV inhibitory activity was determined by the enzymatic assay. Among tested fragments, a potent fragment (LDQWLCEKL), with the half-maximal inhibitory concentration (IC50) of 131 µM was obtained. Further analysis shows that the LDQWLCEKL peptide corresponds to the amino acid sequence of f(115-123) in α-lactalbumin. Furthermore, LDQWLCEKL exhibited a typical non-competitive mode of inhibition. The results indicate that α-lactalbumin contains active peptides with DPP-IV inhibitory activity that may be used to prevent and treat T2D.
Asunto(s)
Dipeptidil Peptidasa 4/metabolismo , Inhibidores de la Dipeptidil-Peptidasa IV/química , Lactalbúmina/metabolismo , Péptidos/química , Proteína de Suero de Leche/metabolismo , Secuencia de Aminoácidos , Animales , Cromatografía Líquida de Alta Presión , Dipeptidil Peptidasa 4/química , Inhibidores de la Dipeptidil-Peptidasa IV/metabolismo , Hidrólisis , Concentración 50 Inhibidora , Cinética , Lactalbúmina/química , Péptidos/análisis , Péptidos/metabolismo , Espectrometría de Masas en Tándem , Tripsina/metabolismoRESUMEN
High chromium yeast has attracted many researchers for its high efficiency and high safety among chromium supplements. The preventive effect of oral high-chromium yeast on diabetes was assessed using KK-AY mice. Sixteen-wk-old type 2 diabetic KK-AY mice were divided into five groups and orally administered with two types of drying processed high-chromium yeast, chromium picolinate at 1000 µg Cr/kg/d, metformin (positive control), and normal yeast (negative control) for 13 weeks. The spray-dried high-chromium yeast significantly delayed the onset of hyperglycemia in type 2 diabetic KK-AY mice (P < 0.05) and significantly improved fasting blood glucose, TG(triglyceride), and TCHO(total cholesterol) . Histopathological analysis showed that the spray-dried high-chromium yeast led to high affinities for the stains to the ß-cells in the islets of Langerhans and alleviated hepatic steatosis. High-chromium yeast could be a potential candidate for nutritional supplement to ameliorate diabetes. PRACTICAL APPLICATION: Chromium plays an important role in fat and carbohydrate metabolism. The result show that spray-dried high chromium yeast significantly delayed the onset of hyperglycemia in type 2 diabetic KK-AY mice. As one of chromium supplements, the purpose of this study is to examine the efficacy of high chromium yeast on the type 2 diabetes and drying method on its bioactivity, which will be useful for research and development of high-chromium yeast and improvement of pharmacological activity-based quality control.
Asunto(s)
Cromo/metabolismo , Diabetes Mellitus Tipo 2/dietoterapia , Hipoglucemiantes/metabolismo , Saccharomyces cerevisiae/metabolismo , Animales , Glucemia/metabolismo , Metabolismo de los Hidratos de Carbono , Cromo/análisis , Diabetes Mellitus Tipo 2/metabolismo , Suplementos Dietéticos/análisis , Modelos Animales de Enfermedad , Humanos , Hipoglucemiantes/análisis , Insulina/sangre , Ratones , Saccharomyces cerevisiae/química , Triglicéridos/metabolismoRESUMEN
A major deterrent to the usage of milk protein concentrate (MPC), a high-protein milk product with increasing demand as a food and sports drink ingredient, has been its poor functional characteristics when compared with other milk protein products such as whey protein concentrate and sodium caseinates. This review discusses the recent research on functional properties of MPC, focusing on factors that may contribute to the poor functional characteristics before, during, and after production. Current research, methods employed, and new understanding on the causes of poor solubility of MPC at mild temperatures (about 20°C) has been presented, including loss of solubility during storage as these areas have received unprecedented attention over the past decade, and also affects other useful functional properties of MPC, such as emulsifying properties, gelation, and foaming. Processing methods, which include heat treatment, high-pressure application, microwave heating, ultrasound application, and enzyme and salts modification, have been used or have potential to modify or improve the functional properties of MPCs. Future research on the effects of these processing methods on the functional properties, including effects of enzyme hydrolysis on bitterness and bioactivity, has also been discussed.
Asunto(s)
Alimentos Funcionales , Proteínas de la Leche/química , Animales , Caseínas , Manipulación de Alimentos/métodos , Conservación de Alimentos , Solubilidad , Gusto , Temperatura , Proteína de Suero de LecheRESUMEN
Autolysis of lactic acid bacteria (LAB) plays a vital role in dairy processing. During cheese making, autolysis of LAB affects cheese flavor development through release of intracellular enzymes and restricts the proliferation of cells in yogurt fermentation and probiotics production. In order to explore the mechanism of autolysis, the gene for the autolytic enzymes of L. bulgaricus, N-acetylmuramidase (mur), was cloned and sequenced (GenBank accession number: KF157911). Mur gene overexpression and gene knockout vectors were constructed based on pMG76e and pUC19 vectors. Recombinant plasmids were transformed into L. bulgaricus ljj-6 by electroporation, then three engineered strains with pMG76e-mur vector and fifteen engineered strains with pUC19-mur::EryBII were screened. The autolysis of the mur knockout strain was significantly lower and autolysis of the mur overexpressed strain was significantly higher compared with that of the wild type strain ljj-6. This result suggested that the mur gene played an important role in autolysis of L. bulgaricus. On the other hand, autolytic activity in a low degree was still observed in the mur knockout strain, which implied that other enzymes but autolysin encoded by mur were also involved in autolysis of L. bulgaricus.
Asunto(s)
Autólisis/genética , Queso/microbiología , Fermentación/genética , Glicósido Hidrolasas/genética , Lactobacillus delbrueckii/genética , Secuencia de Bases , Clonación Molecular , ADN Bacteriano/genética , Técnicas de Inactivación de Genes , Glicósido Hidrolasas/biosíntesis , Lactobacillus delbrueckii/enzimología , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Yogur/microbiologíaRESUMEN
As a highly efficient and easily absorbable source of chromium, the identities of the chromium-binding substances in yeast remain unclear. In this study, a mild extraction procedure involving extraction with ammonia, three-gel filtration, and high-performance liquid chromatography was adopted to obtain two chromium-binding substances from high-chromium yeast. A low-molecular-weight chromium-binding substance was identified, with mass-to-charge ratios (m/z) of 769 and 712, which included glutamic acid, glycine, and cysteine in an approximate ratio of 1:1:1, as well as nicotinic acid and chromium(III). Furthermore, it significantly potentiated (by 51%) the action of insulin to stimulate the conversion of (14)C-glucose into lipid in adipocytes. A novel high-molecular-weight chromium-binding substance was also isolated: electrospray ionization tandem mass spectrometry tentatively identified it as HUB1 target protein-1, glyceraldehyde-3-phosphate dehydrogenase, or ribosomal protein L2A(L5A)(rp8)(YL6). This is the first report of a high-molecular-weight chromium-binding substance in yeast and merits further studies.
Asunto(s)
Proteínas Portadoras/química , Cromo/metabolismo , Proteínas de Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/química , Proteínas Portadoras/aislamiento & purificación , Proteínas Portadoras/metabolismo , Cromo/análisis , Peso Molecular , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/aislamiento & purificación , Proteínas de Saccharomyces cerevisiae/metabolismo , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
The in vivo effects of oral administration of the high-chromium yeast to healthy and diabetic mice are described. Given that these complexes are proposed to function by potentiating the actions of insulin and activating the insulin receptor kinase, changes in lipid and carbohydrate metabolism would be expected. After 15 weeks administration (500 µg Cr/kg body mass) to healthy mice, abnormal metabolism and pathological change were not observed. After 15 weeks of treatment (0-1,000 µg Cr/kg body mass) of diabetic mice, the effect of high-chromium yeast on blood lipids and blood glycosylated hemoglobin (GHb) of diabetes are not consistent. High-chromium yeast results in a lowering (P < 0.05) of GHb and triglyceride, lowering (P < 0.01) of total cholesterol, and restoration (P < 0.01) of insulin; these results are in stark contrast to those of diabetic mice of administration of normal yeast, which have no effect on these parameters and serve as control group. The histopathological analysis of pancreas islet shows that high-chromium yeast could profoundly protect the impaired pancreatic islet and ß-cells from inflammatory infiltration and fibrosis.