Tiliroside, a glycosidic flavonoid, inhibits carbohydrate digestion and glucose absorption in the gastrointestinal tract

Scope Recent studies have reported that tiliroside, a glycosidic flavonoid, possesses anti‐diabetic activities. In the present study, we investigated the effects of tiliroside on carbohydrate digestion and absorption in the gastrointestinal tract. Methods and results This study showed that tiliroside inhibits pancreatic α‐amylase (IC₅₀ = 0.28 mM) in vitro. Tiliroside was found as a noncompetitive inhibitor of α‐amylase with K_i values of 84.2 μM. In male ICR mice, the increase in postprandial plasma glucose levels was significantly suppressed in the tiliroside‐administered group. Tiliroside treatment also suppressed hyperinsulinemia after starch administration. Tiliroside administration inhibited the increase of plasma glucose levels in an oral glucose tolerance test, but not in an intraperitoneal glucose tolerance test. In human intestinal Caco‐2 cells, the addition of tiliroside caused a significant dose‐dependent inhibition of glucose uptake. The inhibitory effects of both sodium‐dependent glucose transporter 1 (SGLT1) and glucose transporter 2 (GLUT2) inhibitors (phlorizin and phloretin, respectively) on glucose uptake were significantly inhibited in the presence of tiliroside, suggesting that tiliroside inhibited glucose uptake mediated by both SGLT1 and GLUT2. Conclusion These findings indicate that the anti‐diabetic effects of tiliroside are at least partially mediated through inhibitory effects on carbohydrate digestion and glucose uptake in the gastrointestinal tract.     

Polyphenols and phenolic acids from strawberry and apple decrease glucose uptake and transport by human intestinal Caco‐2 cells

The effect of polyphenols, phenolic acids and tannins (PPTs) from strawberry and apple on uptake and apical to basolateral transport of glucose was investigated using Caco‐2 intestinal cell monolayers. Substantial inhibition on both uptake and transport was observed by extracts from both strawberry and apple. Using sodium‐containing (glucose transporters SGLT1 and GLUT2 both active) and sodium‐free (only GLUT2 active) conditions, we show that the inhibition of GLUT2 was greater than that of SGLT1. The extracts were analyzed and some of the constituent PPTs were also tested. Quercetin‐3‐O‐rhamnoside (IC₅₀=31 μM), phloridzin (IC₅₀=146 μM), and 5‐caffeoylquinic acid (IC₅₀=2570 μM) contributed 26, 52 and 12%, respectively, to the inhibitory activity of the apple extract, whereas pelargonidin‐3‐O‐glucoside (IC₅₀=802 μM) contributed 26% to the total inhibition by the strawberry extract. For the strawberry extract, the inhibition of transport was non‐competitive based on kinetic analysis, whereas the inhibition of cellular uptake was a mixed‐type inhibition, with changes in both V_max and apparent K_m. The results in this assay show that some PPTs inhibit glucose transport from the intestinal lumen into cells and also the GLUT2‐facilitated exit on the basolateral side.     

Lyophilized Tea Extracts of Brickellia cavanillesii (Asteraceae): In Vitro Characterization of Biological Activity

Lyophilized Brickellia cavanillesii (LBC) tea extracts and identified chemical compounds of LBC were examined using in vitro human carcinoma liver (HepG2) cells with and without fetal bovine serum (FBS). Cells were incubated for 24 h with varying concentrations of FBS and LBC, respectively; cytotoxicity was determined spectrophotometrically using MTT (Formazan 3‐(4,5‐dimethyl‐thiazol‐2‐yl)‐2, 5‐diphenyl‐tetrazolium bromide) assay. Furthermore, the potential hypoglycemic activity of LBC tea extracts was investigated using glucose transport and metabolism proteins biomarkers. FBS (0% to 10%) increased the viability of HepG2 cells steadily with increasing concentration. Possible therapeutic effects of LBC were concentration dependent with and without FBS. The cytotoxicity of 12 identified compounds from the LBC extract suggests that the individual compounds inhibited the proliferation of HepG2 cells differentially and do not reflect the inhibition of the whole aqueous LBC. Western blot analysis of glucose facilitated transporter protein 2 (GLUT 2) expression of HepG2 cells exposed to 0 mg/mL (Control) and 0.2 mg/mL LBC for 2, 4, 6, and 24 h suggests that GLUT 2 expression was increased. Increase in GLUT 2 expression in the absence of FBS was statistically significant with time of exposure. Significant difference was observed for GLUT 2 expression between 6 and 24 h and also between 4 and 24 h at 0.2 mg/mL LBC. Results obtained indicate that LBC may exhibit antidiabetic activity. However, further studies will be necessary to clearly delineate LBC potential therapeutic benefit and biological activities in animal studies as well as other in vitro models.     

Diet effects on glucose absorption in the small intestine of neonatal calves: Importance of intestinal mucosal growth,lactase activity,and glucose transporters

Colostrum (C) feeding in neonatal calves improves glucose status and stimulates intestinal absorptive capacity, leading to greater glucose absorption when compared with milk-based formula feeding. In this study, diet effects on gut growth, lactase activity, and glucose transporters were investigated in several gut segments of the small intestine. Fourteen male German Holstein calves received either C of milkings 1, 3, and 5 (d 1, 2, and 3 in milk) or respective formulas (F) twice daily from d 1 to d 3 after birth. Nutrient content, and especially lactose content, of C and respective F were the same. On d 4, calves were fed C of milking 5 or respective F and calves were slaughtered 2 h after feeding. Tissue samples from duodenum and proximal, mid-, and distal jejunum were taken to measure villus size and crypt depth, mucosa and brush border membrane vesicles (BBMV) were taken to determine protein content, and mRNA expression and activity of lactase and mRNA expression of sodium-dependent glucose co-transporter-1 (SGLT1) and facilitative glucose transporter (GLUT2) were determined from mucosal tissue. Additionally, protein expression of SGLT1 in BBMV and GLUT2 in crude mucosal membranes and BBMV were determined, as well as immunochemically localized GLUT2 in the intestinal mucosa. Villus circumference, area, and height were greater, whereas crypt depth was smaller in C than in F. Lactase activity tended to be greater in C than in F. Protein expression of SGLT1 was greater in F than in C. Parameters of villus size, lactase activity, SGLT1 protein expression, as well as apical and basolateral GLUT2 localization in the enterocytes differed among gut segments. In conclusion, C feeding, when compared with F feeding, enhances glucose absorption in neonatal calves primarily by stimulating mucosal growth and increasing absorptive capacity in the small intestine, but not by stimulating abundance of intestinal glucose transporters.     

Expression and regulation of glucose transporters in the bovine mammary gland

Glucose is the primary precursor for the synthesis of lactose, which controls milk volume by maintaining the osmolarity of milk. Glucose uptake in the mammary gland plays a key role in milk production. Glucose transport across the plasma membranes of mammalian cells is carried out by 2 distinct processes: facilitative transport, mediated by a family of facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Transport kinetic studies indicate that glucose transport across the plasma membrane of the lactating bovine mammary epithelial cell has a K(m) value of 8.29 mM for 3-O-methyl-D-glucose and can be inhibited by both cytochalasin-B and phloretin, indicating a facilitative transport process. This is consistent with the observation that in the lactating bovine mammary gland, GLUT1 is the predominant glucose transporter. However, the bovine lactating mammary gland also expresses GLUT3, GLUT4, GLUT5, GLUT8, GLUT12, and sodium-dependent SGLT1 and SGLT2 at different levels. Studies of protein expression and cellular and subcellular localizations of these transporters are needed to address their physiological functions in the mammary gland. From late pregnancy to early lactation, expression of GLUT1, GLUT8, GLUT12, SGLT1, and SGLT2 mRNA increases from at least 5-fold to several hundred-fold, suggesting that these transporters may be regulated by lactogenic hormones and have roles in milk synthesis. The GLUT1 protein is detected in lactating mammary epithelial cells. Its expression level decreases from early to late lactation stages and becomes barely detectable in the nonlactating gland. Both GLUT1 mRNA and protein levels in the lactating mammary gland are not significantly affected by exogenous bovine growth hormone, and, in addition, GLUT1 mRNA does not appear to be affected by leptin.     

Egg White Ovotransferrin‐Derived ACE Inhibitory Peptide Ameliorates Angiotensin II‐Stimulated Insulin Resistance in Skeletal Muscle Cells

1 Scope

The renin‐angiotensin system (RAS) is a major contributor to the development of insulin resistance and its related complications. Egg white ovotransferrin‐derived tripeptides, IRW (Ile‐Arg‐Trp), IQW (Ile‐Gln‐Trp), or LKP (Leu‐Lys‐Pro) are previously identified as the inhibitors of angiotensin‐converting enzyme (ACE), a key enzyme in the RAS. This study aims at determining whether these peptides are effective in improving insulin resistance, and their mechanisms of action, in a rat derived skeletal muscle cell line (L6 cells).

2 Methods and results

Insulin resistance is induced by treating L6 cells with 1 μm angiotensin II (Ang II) for 24 h. Effects of peptides on glucose uptake are determined using glucose uptake assay, glucose transporter 4 (GLUT4) translocation by immunofluorescence, reactive oxygen species (ROS) by dihydroethidium (DHE) staining, while insulin signaling pathway, Ang II receptor (AT1R or AT2R) levels, and NADPH oxidase activation are measured using Western Blot. Only IRW treatment significantly improves insulin resistance in L6 cells via stimulation of insulin signaling. IRW decreases Ang II‐stimulated AT1R expression, ROS formation, and NADPH oxidase activation.

3 Conclusions

Of three ACE inhibitory peptides studied, only IRW improves insulin resistance in L6 cells, at least partially via reduced AT1R expression and its anti‐oxidative activity.     

In vitro regulation of enzymatic release of glucose and its uptake by Fenugreek microgreen and Mint leaf extract

Fenugreek microgreen and mint leaf displayed antidiabetic potential during in vitro assays including cell line‐based analysis. Aqueous fenugreek microgreen extract (FME) (2 mg mL^?1) inhibited α‐amylase by 70%. It also enhanced glucose uptake in L6 cells by 25% at 10 mg mL^?1 which further improved to 44% in the presence of insulin. On the other hand, fresh mint leaf extract (MLE) inhibited α‐glucosidase up to 90% and increased glucose uptake by 15% in HepG2 cells. Besides, both these extracts also inhibited nonenzymatic glycation of protein. FME and MLE were also found to have high levels of total phenolics, flavonoids and antioxidants which could play a possible role in the observed antidiabetic activity. As blood glucose levels in diabetes can be regulated by inhibition of enzymes regulating carbohydrate metabolism and improving glucose uptake in cells, the findings indicate a complimentary therapeutic role of these two dietary herbs in diabetes.     

Cinnamyl Isobutyrate Decreases Plasma Glucose Levels and Total Energy Intake from a Standardized Breakfast: A Randomized,Crossover Intervention

1 Scope

Cinnamon is associated with anti‐obesity effects, regulating food intake, improving plasma glucose levels and lipid profiles in vivo. In the present study, the impact of cinnamyl isobutyrate (CIB), one constituent of cinnamon, on ad libitum food intake from a standardized breakfast and outcome measures of hormonal regulation of appetite were investigated.

2 Methods and results

In this randomized, short‐term crossover intervention study, a 75 g per 300 mL glucose solution solely (control) or supplemented with 0.45 mg CIB was administered to 26 healthy volunteers. Prior to and 2 h after receiving control or CIB treatment, subjective hunger perceptions were rated using a visual analog scale. Food intake from a standardized breakfast was assessed 2 h after treatments. Plasma peptide YY_3–36, glucagon‐like‐peptide1, ghrelin, and serotonin as well as plasma glucose and insulin were measured in blood samples drawn at fasting and 15, 30, 60, 90, and 120 min after treatment. CIB administration decreased total energy intake and delta area under curve plasma glucose by 4.64 ± 3.51% and 49.3 ± 18.5% compared to control treatment, respectively.

3 Conclusions

CIB, administered at a 0.45 mg bolus in 75 g glucose–water solution, decreased ad libitum energy intake from a standardized breakfast and postprandial plasma glucose levels.     

Antioxidant and tyrosinase inhibitory activities of different parts of guava (<Emphasis Type="Italic">Psidium guajava</Emphasis> L.)

The antioxidant and the tyrosinase inhibitory activities of 4 different solvents (acetone, ethanol, methanol, and water) for preparation of extracts from guava (branch, fruit, leaf, and seed) were evaluated by measuring total phenolic contents (TPC), DPPH radical scavenging activity, ABTS radical scavenging activity, reducing power (RP), and tyrosinase inhibitory activity. The extracts of branch and leaf showed relatively higher antioxidant properties than those of fruit and seed. The highest TPC (141.28 mg/g gallic acid equivalents), DPPH radical scavenging activity (IC₅₀=34.01 μg/mL), ABTS radical scavenging activity (IC₅₀=3.23 μg/mL), and RP (IC₅₀= 75.63 μg/mL) were found in acetone extract of leaf, while water extract of seed had the lowest antioxidant activity. The tyrosinase inhibitory activity of ethanol extract from guava leaf was 69.56%, which was the highest activity among the extracts. These results indicate that useful bioactive substances exist in the guava branch as well as leaf extracts.     

Effect of Guava (Psidium guajava L.) Leaf Extract on Glucose Uptake in Rat Hepatocytes

ABSTRACT:  People in oriental countries, including Japan and Taiwan, boil guava leaves ( Psidium guajava L.) in water and drink the extract as a folk medicine for diabetes. The present study investigated the enhancement of aqueous guava leaf extract on glucose uptake in rat clone 9 hepatocytes and searched for the active compound. The extract was eluted with MeOH-H₂O solutions through Diaion, Sephadex, and MCI-gel columns to separate into fractions with different polarities. The uptake test of 2-[1-¹⁴C] deoxy-D-glucose in rat clone 9 hepatocytes was performed to evaluate the hypoglycemic effect of these fractions. The active compound was identified by nuclear magnetic resonance analysis and high-performance liquid chromatography (HPLC). The results revealed that phenolics are the principal component of the extract, that high polarity fractions of the guava leaf extract are enhancers to glucose uptake in rat clone 9 hepatocytes, and that quercetin is the major active compound. We suggest that quercetin in the aqueous extract of guava leaves promotes glucose uptake in liver cells, and contributes to the alleviation of hypoglycemia in diabetes as a consequence.     

Short communication: Protein kinase C regulates glucose uptake and mRNA expression of glucose transporter (GLUT) 1 and GLUT8 in lactating bovine mammary epithelial cells

The aim of this study was to determine the role of protein kinase C (PKC) in regulating glucose uptake in lactating bovine mammary epithelial cells (BMEC). The BMEC were cultured and treated with different concentrations of phorbol 12-myristate 13-acetate (PMA;0, 10, 25, 50, 100, and 200 ng/mL), the classic activator of PKC, for 48 h. Compared with the cells with no PMA treatment, 50 and 100 ng of PMA/mL significantly stimulated the glucose uptake of the BMEC, whereas the glucose uptake by the cells treated with the lowest and the highest amounts of PMA (25 and 200 ng/mL, respectively) did not show a significant difference. Consistently, the mRNA expression of glucose transporter (GLUT) 1 and 8 showed a similar pattern of increase under the treatments of PMA. Furthermore, when the cells were pretreated with GF1090203X (0, 0.25, 0.5, 1, and 2 μM), an inhibitor of PKC, for 30 min before exposed to PMA (50 ng/mL), the PMA-induced glucose uptake and GLUT1 and GLUT8 expression were decreased by GF1090203X in a dose-dependent manner. These results demonstrate that PKC is involved in the regulation of glucose uptake by BMEC, and this function may work, at least partly, through upregulating the expression of GLUT1 and GLUT8.     

Prenylated chalcones 4-hydroxyderricin and xanthoangelol stimulate glucose uptake in skeletal muscle cells by inducing GLUT4 translocation

Scope: Glucose uptake in skeletal muscle is crucial for glucose homeostasis. Methods and results: Insulin and muscle contraction increase glucose uptake accompanied by the translocation of glucose transporter (GLUT) 4. In a search for promising foods, which can increase glucose uptake in skeletal muscle, we screened for active polyphenols by assaying for uptake of 2‐deoxyglucose (2DG) in rat L6 muscle cells. Among 37 compounds, 4‐hydroxyderricin and xanthoangelol, prenylated chalcones abundant in Ashitaba (Angelica keiskei Koidzumi, family Apiaceae), significantly increased 2DG uptake in L6 cells by 1.9‐fold at 10 μM, compared with the level in DMSO‐treated control cells. Next, we investigated the effect of these chalcones on the translocation of GLUT4 and its underlying mechanisms. The chalcones increased the GLUT4 level in the plasma membrane of L6 cells, but activated neither protein kinase C ζ/λ, Akt, nor adenosine monophosphate‐activated protein kinase, all of which regulate the GLUT4 translocation. Interestingly, the oral administration of a titrated chalcone‐enriched Ashitaba extract containing 150.6 mg/g (dry base) of 4‐hydroxyderricin and 146.0 mg/g (dry base) of xanthoangelol suppressed acute hyperglycemia in oral glucose tolerance tests of mice. Conclusions: Ashitaba is a promising functional food for the maintenance of the blood glucose level by inducing skeletal muscle‐associated glucose uptake.     

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.     

花椒精油对链脲佐菌素诱导的糖尿病小鼠糖代谢的影响

目的:研究花椒精油对1型糖尿病机体糖代谢的影响及可能机制.方法:将50只健康雄性昆明小鼠(KM小鼠)分为空白对照,糖尿病模型组,花椒精油高(15?mg/(kg mb·d))、中(9?mg/(kg mb·d))、低(3?mg/(kg mb·d))剂量组,每组10只.采用链脲佐菌素诱导建立1型糖尿病模型.空白组和糖尿病模型...     

Anti-inflammatory effects of 4 medicinal plant extracts in lipopolysaccharide-induced RAW 264.7 cells

The anti-inflammatory activity of 4 plant extracts [guava (Psidium guajava) leaf, capillary wormwood (Artemisia capillaris Thunb.), Chinese goldthread (Coptis chinensis), and dandelion (Taraxacum platycarpum)] was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Six phenolic compounds (gallic acid, caffeic acid, chlorogenic acid, catechin, quercetin, and baicalin) were analyzed using LC-MS/MS. Guava leaf extracts showed the highest inhibitory effects on LPS-induced nitric oxide (NO, 52.58%) and prostaglandin E₂ (PGE₂, 43.45%) production. The total phenolic contents (TPC) in guava leaf, capillary wormwood, Chinese goldthread, and dandelion were 426.84, 154.42, 41.73, and 122.04 mg of gallic acid equivalent (GAE)/g of extract, respectively. TPC was positively correlated with the NO-inhibitory effect (r= 0.963, p<0.05) and the PGE₂-inhibitory effect (r=0.971, p<0.05) at 30 μg/mL of treatment. The guava leaf extracts contained the highest levels of gallic acid and catechin, while the capillary wormwood extracts contained the highest levels of chlorogenic acid and quercetin.