Antidiabetic Potential of Green Seaweed Enteromorpha prolifera Flavonoids Regulating Insulin Signaling Pathway and Gut Microbiota in Type 2 Diabetic Mice

This study aimed to investigate the antidiabetic activity of water–ethanol extract of green macroalgae Enteromorpha prolifera (EPW) and its flavonoid‐rich fraction less than 3 kDa (EPW3) in type 2 diabetic mice induced by streptozotocin and a high‐sucrose/high‐fat diet. The major active compounds were identified using ultra‐performance liquid chromatography coupled with quadrupole‐time‐of‐flight‐tandem mass spectrometry. Quantitative gene expression analysis of the insulin signaling pathway was performed. The effects of EPW3 on gut microflora in diabetic mice were analyzed by high‐throughput 16S rRNA gene sequencing. The results showed EPW3 treatment decreased the fasting blood glucose, improved oral glucose tolerance, and protected against liver and kidney injury with reduced inflammation in diabetic mice. The active principle of EPW3 revealed hypoglycemic effect as indicated by activation of the IRS1/PI3K/AKT and inhibition of the JNK1/2 insulin pathway in liver. Furthermore, the treatment significantly enriched the abundance of Lachnospiraceae and Alisties, which were positive correlation of metabolic phenotypes. These findings indicated that EPW3 possessed great therapeutic potential as adjuvant therapy for type 2 diabetes.     

Lipid metabolic effect of Korean red ginseng extract in mice fed on a high-fat diet

BACKGROUND: Ginseng saponin and ginsenosides exert anti‐obesity effects via the modulation of physiological lipid metabolism in vivo or intracellular signalling in cell culture systems. However, the complicated relationship between the anti‐obesity effects of ginseng and gene expression has yet to be defined under in vivo conditions. Therefore, we evaluated the relationship between the anti‐obesity effects of Korean red ginseng extract (KRGE) and hepatic gene expression profiles in mice fed long‐term on a high‐fat diet (HFD) in this study. RESULTS: KRGE reduces the levels of cholesterol, low‐density lipoprotein‐cholesterol (LDL‐C), serum triglycerides, and atherogenic indices. Levels of leptin, adiponectin and insulin, which regulate glucose and lipid metabolism, were impaired profoundly by HFD. However, KRGE treatment brought these levels back to normal. KRGE was found to down‐regulate genes associated with lipid metabolism or cholesterol metabolism (Lipa, Cyp7a1, Il1rn, Acot2, Mogat1, Osbpl3, Asah3l, Insig1, Anxa2, Vldlr, Hmgcs1, Sytl4, Plscr4, Pla2g4e, Slc27a3, Enpp6), all of which were up‐regulated by HFD. CONCLUSION: KRGE regulated the expression of genes associated with abnormal physiology via HFD. Leptin, insulin, and adiponectin, which carry out critical functions in energy and lipid metabolism, were shown to be modulated by KRGE. These results show that KRGE is effective in preventing obesity. Copyright © 2011 Society of Chemical Industry     

Salicornia Extract Ameliorates Salt‐Induced Aggravation of Nonalcoholic Fatty Liver Disease in Obese Mice Fed a High‐Fat Diet

High‐fat and high‐salt intakes are among the major risks of chronic diseases including obesity, nonalcoholic fatty liver disease (NAFLD), and nonalcoholic steatohepatitis (NASH). Salicornia is a halophytic plant known to exert antioxidant, antidiabetic, and hypolipidemic effects, and Salicornia‐extracted salt (SS) has been used as a salt substitute. In this study, the effects of SS and purified salt (PS) on the aggravation of NAFLD/NASH were compared. C57BL/6J male mice (8‐wk‐old) were fed a high‐fat diet (HFD) for 6 mo and divided into 3 dietary groups, which were additionally fed HFD, HFD + SS, and HFD + PS for 13 wk. PS induced aggravation of NAFLD/NASH in HFD‐fed mice. Although the actual salt intake was same between the PS and SS groups as 1% of the diet (extrapolated from the World Health Organization [WHO] guideline), SS induced less liver injury and hepatic steatosis compared to PS. The hepatic mRNA expressions of inflammatory cytokines and fibrosis marker were significantly lower in the SS group than the PS group. Oxidative stress is one of the major causes of inflammation in NAFLD/NASH. Results of the component analysis showed that the major polyphenols that exhibited antioxidant activity in the Salicornia water extract were ferulic acid, caffeic acid, and isorhamnetin. These results suggest that even the level of salt intake recommended by WHO can accelerate the progression of liver disease in obese individuals consuming HFD. It is proposed that SS can be a salt substitute for obese individuals who consume HFD.     

Salvianolic Acid B Inhibits High‐Fat Diet‐Induced Inflammation by Activating the Nrf2 Pathway

Salvianolic acid B (Sal B) is a major water‐soluble bioactive component of Salvia miltiorrhiza, which is a traditional Chinese medicine. We investigated the ways in which Sal B affects high‐fat diet (HFD)‐induced immunological function disorder remission using a C57BL/6 mouse model. We gave groups of C57BL/6 mice a normal diet (Control), a normal diet supplemented with Sal B (Control + Sal B), a high‐fat diet (HF), and a high‐fat diet supplemented with Sal B (HF + Sal B) for 10 wk. Sal B supplementation decreased the body weight and plasma lipids, increased the fecal excretion of lipids, prevented the accumulation of chronic oxidative stress, and reversed the disproportionality of CD3⁺CD4⁺ and CD3⁺CD8⁺ T lymphocytes compared to HFD. We found an increase in IL‐6 and TNF‐α, while IL‐10 decreased in plasma after the HFD and Sal B reversed the deregulation of the Thl/Th2 ratio. In addition, HFD‐induced inflammation was stopped by Sal B through the downregulation of nuclear factor‐κB (NF‐κB), cyclooxygenase‐2 (COX‐2), and inducible NO synthesis (iNOS), and the upregulation of nuclear factor‐erythroid 2‐related factor 2 (Nrf2)‐regulated genes. These findings demonstrated that Sal B could effectively attenuate inflammation by activating the Nrf2‐mediated antioxidant defense system.     

LC-MSn study of the chemical transformations of hydroxycinnamates during yerba maté (Ilex paraguariensis) tea brewing

Yerba maté is one of the most popular beverages in South American countries and its consumption is associated with a wide array of health effects. In this study, we used advanced HPLC-ESI-MSⁿ and HPLC-ESI-HRMS methods for the identification and characterization of hydroxycinnamates and their derivatives formed during the brewing process of yerba maté. We report on the hydroxylation of the hydroxycinnamates cinnamoyl substituent by conjugate addition of water to form 3-hydroxy-dihydrocinnamic acid derivatives using a series of model compounds, including caffeoylglucoses, dicaffeoylquinic acids, methyl caffeoylquinate and mono caffeoylquinic acids. The regiochemistry of conjugate addition was determined by targeted tandem MS experiments performed on authentic standards. It was interesting to note that hydroxylation of hydroxycinnamates produced cis and acyl-migration isomers, which is in line with previously reported data.     

The in vitro and in vivo effects of yerba mate (Ilex paraguariensis) extract on adipogenesis

The aim of this study was to evaluate the effects of yerba mate extract and its principal bioactive compounds on adipogenesis. The anti-adipogenic effects of yerba mate, chlorogenic acid, quercetin and rutin were evaluated in 3T3-L1 cells using a PCR array. The results obtained in vitro were validated in vivo in a high-fat diet-induced model of obesity. The in vitro and in vivo results demonstrated that yerba mate extract down-regulated the expression of genes that regulate adipogenesis, such as Creb-1and C/EBPα, and the extract up-regulated the expression of genes related to the inhibition of adipogenesis, including Dlk1, Gata2, Gata3, Klf2, Lrp5, Pparγ₂, Sfrp1, Tcf7l2, Wnt10b, and Wnt3a. In summary, it was demonstrated that yerba mate and its bioactive compounds regulate the expression of genes related to in vitro adipogenesis. Furthermore, yerba mate might regulate adipogenesis through the Wnt pathway.     

Micronutrients in High-Fat Diet Modify Insulin Resistance and Its Regulatory Genes in Adult Male Mice

Scope

Obesity and insulin resistance (IR) are associated with epigenetic changes of gene expression. However, the relationship between micronutrients, epigenetic regulation of gene expression, and IR during development of diet-induced obesity has yet to be defined. Our objective is to describe the effect of micronutrient addition to diets on IR and its related genes during obesity development.

Methods and results

Male C57BL/6J mice are fed a high-fat (HFD) or low-fat (LFD) diets with or without a multi-vitamin mineral mix (MVM) addition containing vitamins A, B1, B6, B12, and Zn, and Se for 9 weeks. Compared to LFD mice, HFD mice have higher body weight, IR, fasting glucose, insulin, C-peptide, leptin, and hepatic triglyceride concentrations, and dysregulated gene expression in liver, muscle, pancreas, and fat tissues (p < 0.05). The addition of MVM reduces these HFD-induced effects. HFD downregulates 27 genes associated with insulin regulation and adipose tissue function across all tissues by an average of 47% and upregulates five genes by 230% (p < 0.001). Adding MVM downregulates five genes and upregulates one in HFD-fed mice. Both HFD and MVM alter one-carbon metabolites.

Conclusion

Addition of micronutrients to the HFD decreases IR and modifies associated gene expression in obese and lean mice.     

Chlorogenic acid promotes osteoblastogenesis in human adipose tissue-derived mesenchymal stem cells

High-fat diet (HFD)-induced obesity is associated with oxidative stress. The purpose of this study was to examine the antioxidant effect of Phaeodactylum tricornutum extract in mice with diet-induced obesity. Four-week-old C57BL/6J mice were fed a normal diet or HFD with and without 0.7% P. tricornutum lipid extract corresponding to 0.2% fucoxanthin for 8 weeks. P. tricornutum significantly decreased body weight and epidydimal white adipose tissue in mice fed the HFD. Serum triglyceride, glucose, insulin, and leptin levels, as well as homeostasis model assessment for insulin resistance (HOMA-IR) values, were significantly lower in the P. tricornutum group than in the HFD group. P. tricornutum significantly decreased thiobarbituric acid reactive substances (TBARS) and increased glutathione and the activities of superoxide dismutase, catalase, and glutathione peroxidase in the liver compared with the HFD group. Thus, P. tricornutum could exert antiobesity and antioxidant effects in mice fed a HFD.     

豌豆低聚肽对2型糖尿病小鼠肝脏PI3K/AKT/FOXO1信号通路的调节作用

本文旨在探究豌豆低聚肽对2型糖尿病(T2MD)小鼠肝脏磷脂酰肌醇-3激酶/蛋白激酶B/转录因子FoxO1(PI3K/AKT/FOXO1)蛋白表达的影响.通过对小鼠腹腔注射链脲佐菌素(STZ),建立2型糖尿病小鼠模型,并用二甲双胍和豌豆低聚肽分别饮食干预4周.结果表明,豌豆低聚肽组的小鼠糖尿病指征较模型组均有一定改善,且...     

Green tea polyphenols reduce obesity in high‐fat diet‐induced mice by modulating intestinal microbiota composition

In this study, the modulatory effect of green tea polyphenols (GTP) on human intestinal microbiota was investigated. Firstly, germ‐free mice were inoculated with faecal suspension derived from healthy volunteers to obtain human flora‐associated (HFA) mice model. When the high‐fat diet‐induced obese mice model was successfully established, they were randomly divided into high‐fat diet group (HFD) and high‐fat diet group with GTP (HFD‐GTP), and the shifts in relative abundance of the dominant taxa at the phylum, family and genus levels were studied by high‐throughput sequencing. The diversity of the total bacterial community reached the maximum after GTP treatment for 3 weeks, and then decreased when the mice were fed without GTP. Despite interindividual variation, the relative abundance of Bacteroidetes increased from 0.56 ± 0.06 (1st week) to 0.60 ± 0.05 (3th week), while Firmicutes decreased from 0.42 ± 0.06 to 0.37 ± 0.02. Interestingly, certain bacterial communities such as Bacteroidetes and Proteobacteria still increased and Firmicutes showed a decreasing trend when the mice were fed without GTP (4th week). The results showed that GTP benefits the stability of certain gut microbiota, especially in an environment‐triggered microbial imbalance; therefore, it may have prebiotic‐like activity contributing to the prevention of obesity.     

Extracellular Vesicles from Hypoxic Adipocytes and Obese Subjects Reduce Insulin‐Stimulated Glucose Uptake

1 Scope

We investigate the effects of extracellular vesicles (EVs) obtained from in vitro adipocyte cell models and from obese subjects on glucose transport and insulin responsiveness.

2 Methods and results

EVs are isolated from the culture supernatant of adipocytes cultured under normoxia, hypoxia (1% oxygen), or exposed to macrophage conditioned media (15% v/v). EVs are isolated from the plasma of lean individuals and subjects with obesity. Cultured adipocytes are incubated with EVs and activation of insulin signalling cascades and insulin‐stimulated glucose transport are measured. EVs released from hypoxic adipocytes impair insulin‐stimulated 2‐deoxyglucose uptake and reduce insulin mediated phosphorylation of AKT. Insulin‐mediated phosphorylation of extracellular regulated kinases (ERK1/2) is not affected. EVs from individuals with obesity decrease insulin stimulated 2‐deoxyglucose uptake in adipocytes (p = 0.0159).

3 Conclusion

EVs released by stressed adipocytes impair insulin action in neighboring adipocytes.     

Diet‐induced obesity regulates the galanin‐mediated signaling cascade in the adipose tissue of mice

Galanin is a neuropeptide that regulates the food intake, neurogenesis, memory, and gut secretion. This study was conducted to evaluate the high‐fat diet (HFD)‐induced regulation of the galanin receptors (GalRs) and the associated signaling molecules in the adipose tissues of mice. Twenty C57BL/6J mice were given either an HFD or a normal diet for 12 wk. The results of the semiquantitative RT‐PCR analyses indicated that the HFD upregulated the expression of GalR1, GalR2, GalR3, resistance to audiogenic seizures, peroxisome proliferator‐activated receptorγ2, adipocyte protein 2, and protein kinase Cδ and downregulated the expression of peroxisome proliferative activated receptor γ coactivator 1α and uncoupling protein 1 in the adipose tissues. The immunoblot results showed that the protein levels of peroxisome proliferator‐activated receptorγ2 and adipocyte protein 2, and the phosphorylation of c‐Raf and extracellular signal‐regulated kinase 1/2 were increased, while the phosphorylation of cyclic adenosine monophosphate‐responsive element‐binding protein, which regulates peroxisome proliferative activated receptor γ coactivator 1α and uncoupling protein 1, was decreased in the epididymal adipose tissues of the HFD‐fed mice. These results suggest the possible association of the galanin‐mediated signaling pathways in the manifestation of the HFD‐induced activation of adipogenesis along with the suppression of thermogenesis in the adipose tissues of mice.     

Antioxidant kinetics of plant-derived substances and extracts

The antioxidant activity (AA) of substances present in several plant species has been widely studied which reflects their fundamental role in the protection of skin tissue against the harmful action of reactive oxygen species. Given the importance of effective and long‐lasting protection against ultraviolet radiation, we studied the AA of several plant derivatives and extracts over time. Several chemical in vitro methods may be used to evaluate antioxidant capability, among which the 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) method stands out, despite its unspecificity, as the most cited and described method in the literature. In this work the AA was evaluated by measuring their capacity to reduce DPPH in 30 min, which is suggested in the literature, and additionally at different times up to 8 h from the baseline reading. The methodology used to evaluate the AA over time was validated. It is important to emphasize that this study proposes to modify the conventional DPPH method, although considered to be non‐specific, to be used to test new antioxidant agents. This represents a considerable advantage because some substances show no significant activity during the first 30 min of reaction. Among other plant products, we tested a proantocyanidin‐rich grapeseed extract, a hesperidin derivative, a rutin‐containing ginkgo extract, a polyphenol‐containing yerba maté extract and tocopheryl acetate, all of which were properly standardized. As they have different antioxidant profiles, each ingredient showed a specific behaviour over time, which may promote the selection of anti‐radical compounds capable of offering protection against external agents. Combining extracts and plant derivatives that present fast, medium and slow antioxidant kinetic it is possible to create complexes capable of offering an effective protection from the moment of application up to several hours later. It is a perfectly feasible method, and such combinations prove to be more effective and have more durable effect.     

High‐fat Diet‐induced Intestinal Hyperpermeability is Associated with Increased Bile Acids in the Large Intestine of Mice

Metabolic syndrome is characterized by low‐grade chronic systemic inflammation, which is associated with intestinal hyperpermeability. This study examined the effects of 3 high‐fat diets (HFDs) composed of different fat sources (soybean oil and lard) on the intestinal permeability, tight junction (TJ) protein expression, and cecal bile acid (BA) concentrations in mice, and then analyzed their interrelations. C57/BL6 mice were fed the control diet, HFD (soybean oil), HFD (lard), and HFD (mix; containing equal concentrations of soybean oil and lard) for 8 wk. Glucose tolerance, intestinal permeability, TJ protein expression, and cecal BA concentration were evaluated. Feeding with the 3 HDFs similarly increased body weight, liver weight, and fat pad weight, and induced glucose intolerance and intestinal hyperpermeability. The expression of TJ proteins, zonula occludens‐2 and junctional adhesion molecule‐A, were lower in the colons of the 3 HFD groups than in the control group (P < 0.05), and these changes appeared to be related to intestinal hyperpermeability. Feeding with HFDs increased total secondary BA (SBA) and total BA concentrations along with increases in some individual BAs in the cecum. Significant positive correlations between intestinal permeability and the concentrations of most SBAs, such as deoxycholic acid and ω‐muricholic acids, were detected (P < 0.05). These results suggest that the HFD‐induced intestinal hyperpermeability is associated with increased BA secretion. The abundance of SBAs in the large intestine may be responsible for the hyperpermeability.     

Decreased fat accumulation in 3T3‐L1 pre‐adipocytes treated with extracts of heat‐processed soy flour and breads

The potential anti‐adipogenic effects of gluten‐free soy breads made from germinated soybean (GS), steamed soybean (SS) or roasted soybean (RS) were evaluated in an in vitro adipocyte cell model. GS and RS increased the total phenolic (TP) and total flavonoid (TF) contents of flours compared with the raw soybean (NS) flour. RS and GS had the highest TP (1.04 GAE mg g^?1) and TF (0.92 CAE mg g^?1) contents. Baking increased the TP content of breads, 0.09–0.26 GAE mg g^?1, compared with the flours. Fermentation during breadmaking increased the DPPH scavenging activities compared with the flours. The ABTS scavenging exhibited similar patterns to those of DPPH. Lipid accumulation in 3T3‐L1 cells shows that the alcoholic extracts (100 μg mL^?1) of SS flour and bread decreased adipocyte differentiation by 1.6‐ and 2.1‐fold, respectively, compared with control. SS bread extract substantially downregulated the adipogenesis‐related genes such as acetyl‐CoA carboxylase and glycerol‐3‐phosphate dehydrogenase.     

Inhibitory effects of Hibiscus sabdariffa L extract on low‐density lipoprotein oxidation and anti‐hyperlipidemia in fructose‐fed and cholesterol‐fed rats

Hibiscus sabdariffa L extract (HSE) is an aqueous extract of Hibiscus sabdariffa L flowers that is used as a local soft drink and medical herb in Taiwan. Oxidation of low‐density lipoprotein (LDL) has been shown to increase the incidence of atherosclerosis. In this study, we determined the antioxidative activity of HSE on LDL oxidation by examining relative electrophoretic mobilities (REM) and thiobarbituric acid‐reactive substances (TBARS). The data revealed an inhibitory effect of HSE on Cu²⁺‐mediated REM and TBARS. HSE exhibited a remarkable ability to reduce cholesterol degradation and ApoB fragmentation. Overall, HSE showed a high potency to inhibit the production of oxidized LDL induced by copper and, specifically, to reduce serum triglycerides in high‐fructose diet (HFD) fed rats and serum cholesterol in high‐cholesterol diet (HCD) fed animals. The levels of LDL and the ratio of LDL‐cholesterol (LDL‐C) to HDL‐cholesterol (HDL‐C) were reduced by HSE in both hyperlipidaemia models. Based on these findings, we suggest that HSE may be used to inhibit LDL oxidation and to prevent various types of hyperlipidaemia in HFD‐ or HCD‐fed rats. Copyright © 2004 Society of Chemical Industry     

Integrated Remodeling of Gut–Liver Metabolism Induced by Moderate Protein Restriction Contributes to Improvement of Insulin Sensitivity

1 Scope

Protein restriction (PR) is beneficial for relieving metabolic disorders and aging‐related diseases. However, extreme PR could result in malnutrition due to severe deficiency of essential amino acids. Therefore, the effect of moderate PR on insulin sensitivity is investigated.

2 Methods and results

The growing and adult pigs are subjected to moderate PR by 15–30%. Plasma insulin concentration and insulin resistance index HOMA‐IR are significantly decreased upon moderate PR. Furthermore, IRS1/PI3K/AKT pathway in the basal state is enhanced in both liver and skeletal muscle. The adapted metabolism in the liver upon moderate PR is in support of improving insulin sensitivity. The liver shares a coordinated metabolic adaption in terms of energy metabolism and amino acid metabolism with the small intestine. Particularly, alteration of the metabolic footprint appeared in the portal venous blood, representing metabolites to be absorbed into liver after intestinal metabolism, is also in favor of improvement of insulin sensitivity.

3 Conclusion

In summary, the study proves that moderate PR could improve insulin sensitivity from childhood to adulthood in a pig model, and sheds a new light on the role of integrated remodeling of gut and liver metabolism in the improved insulin sensitivity induced by moderate PR.