Improved Glucose Consumption Research Articles

Linarin Identified as a Bioactive Compound of Lycii Cortex Ameliorates Insulin Resistance and Inflammation Through the c‐FOS/ARG2 Signaling Axis

ABSTRACTInsulin resistance (IR) is a central pathophysiological process underlying numerous chronic metabolic disorders, including type 2 diabetes and obesity. Lycii Cortex, a widely used traditional Chinese herb, has demonstrated potential benefits in preventing and managing diabetes and IR. Whereas, the specific bioactive compounds responsible for these protective effects and their underlying mechanisms of action remain elusive. This study aimed to identify the bioactive components within Lycii Cortex that contribute to its anti‐diabetic effects and to elucidate the molecular mechanisms underlying its beneficial actions on insulin resistance. Network pharmacology and molecular docking analyses were employed to identify the potential active compounds in Lycii Cortex and their corresponding target proteins. An in vitro model of IR was established using palmitic acid (PA)‐treated HepG2 cells. Cell viability was assessed using the CCK‐8 assay, while glucose uptake was evaluated by 2‐NBDG staining and extracellular glucose measurement. To validate the in vitro findings, an in vivo model of obesity‐induced IR was established using high‐fat diet (HFD)‐fed mice. The network pharmacology analysis preliminarily identified 13 candidate chemicals and 10 hub LyC and IR‐related genes (LIRRGs). Molecular docking analysis demonstrates that Linarin as the potential active component exhibits the greatest potential to target c‐FOS for preventing obesity‐induced IR. Enrichment analysis suggested that Linarin‐targeted pathways are correlated with inflammation. In vitro experimental validation demonstrated that Linarin was capable of protecting against PA‐induced IR in HepG2 cells evidenced by improving glucose uptake ability and reducing extracellular glucose content. Additionally, we found that Linarin ablated PA‐induced increase in the expression of c‐FOS and inflammatory cytokines. Furthermore, in PA‐treated cells, silencing c‐FOS markedly improved glucose consumption, and reduced inflammation and Arginase 2 (ARG2) expression. Similarly, as exposure to PA, silencing ARG2 also ameliorated glucose uptake and inflammation, while not affecting c‐FOS expression. In vivo experiments further showed that Linarin administration remarkably improved glucose tolerance and insulin sensitivity, and reduced the fat mass and body weight in HFD‐induced obese mice. In this study, Linarin has been identified as the bioactive compound of Lycii Cortex to ameliorate obesity‐related IR and inflammation through the c‐FOS/ARG2 signaling cascade. These findings underscore the therapeutic potential of Linarin and provide valuable insights into developing novel intervention strategies for type 2 diabetes therapy.

Revealing the potential hypoglycaemic ingredients of Terminalia chebula Retz. by spectrum–effect relationship combining molecular docking and experimental validation

Terminalia chebula Retz. is commonly used in the treatment of diabetes, but its specific ingredients and hypoglycaemic mechanisms remain unclear. In this study, spectrum-effect relationships between common peaks of fingerprint and DPPH, ABTS, PTP1B, α-glucosidase, were used to screen six compounds as the potential hypoglycemic ingredients. Molecular docking studies were conducted on chebulagic acid, chebulinic acid and punicalagin, which were identified by reference standard, revealing the potent in teractions between these bioactive substances and the PTP1B, α-glucosidase protein. Additionally, the cellular level results showed that punicalagin displayed stronger inhibition of PTP1B and α-glucosidase, with IC50 values of 0.0236 and 0.335 μg/mL, respectively, while improving glucose consumption and intracellular glycogen content of insult-resistant L6 muscle cell, and the hypoglycemic effect was achieved by activating the PI3K/AKT signaling pathway. This study demonstrated that T. chebula has the potential to become a hypoglycemic functional food, and offers a strategy for characterization hypoglycemic ingredients.

Tea seed saponin‑reduced extract ameliorates palmitic acid‑induced insulin resistance in HepG2 cells

Tea seed saponin‑reduced extract ameliorates palmitic acid‑induced insulin resistance in HepG2 cells

Structural characterization, and in vitro hypoglycemic activity of a polysaccharide from the mushroom Cantharellus yunnanensis

A polysaccharide CY-2 from C. yunnanensis was obtained through a process of consecutive water extraction, alcohol precipitation, and DEAE-52 fast-flow chromatography. CY-2, with an average molecular weight of 2.69 × 104 Da mainly consisted of glucose and mannose with a molar ratio of 33.5: 56.9. Infrared spectrum (IR), methylation analysis, and nuclear magnetic resonance (NMR) results revealed that CY-2 may have a backbone consisting of →6)-α-D-Manp-(1 → 3)-β-D-Glcp-(1→, and branch chain β-D-Glcp-(1→. Meanwhile, CY-2 had a higher inhibition rate on α-glucosidase activity compared with other fractions (CY-0, CY-1, and CY-4) and was a mixed competitive inhibitor. In addition, CY-2 at the concentration of 10 μg/mL presented a superior power to improve glucose consumption and metabolism in HepG2 cells compared with metformin. Overall, these findings highlight the potential value of CY-2 as a hypoglycemic agent.

Two new macrocarpene-type sesquiterpenes from the styles and stigmas of Zea mays

Two new sesquiterpenes, named zeaols A (1) and B (2), along with three known related analogues, were isolated from the styles and stigmas of Zea mays. Their structures and absolute configurations were elucidated by extensive spectroscopic analyses including UV, 1D- and 2D-NMR, HR-ESI-MS and ECD. Compounds 1 and 2 showed weak cytotoxic activities against the T-24 cancer cell line and compounds 1–5 improved glucose consumption in HepG2 cells.

Hypoglycemic activity of phenols from Pleioblastus amarus (Keng) shells and its main chemical constituents identificatied using UHPLC-Q-TOF-MS

• Pleioblastus amarus (Keng) shells was used as raw material to extract and separate polyphenols (PASP). • The effects of PASP on glucose consumption, intracellular glycogen content, hexokinase and pyruvate kinase activities in insulin-resistant HepG2 cells were measured. • Main chemical constituents of PASP was identified by UHPLC-Q-TOF, and 12 kinds of compounds were identified. • Four kinds of potential α-glucosidase inhibitors were screened by ultrafiltration affinity mass spectrometry. The aim of this study was to investigate the hypoglycemic activity of phenols extracted from Pleioblastus amarus (Keng) shells (PASs), and to isolate and identify the possible components.of which o-coumaric acid, ferulic acid, quercetin-3-0-rhamnoside, and apigenin-7-0-glucoside exhibited a strong affinity with α-glucosidase and were identified as potential The results revealed that, Pleioblastus amarus (Keng) shell phenols (PASP) displayed hypoglycemic activity in vitro, inhibiting α-glucosidase (IC50 = 0.158 ± 0.002 mg/mL) while improving glucose consumption in insulin-resistant (IR) HepG2 cells, as well as the intracellular glycogen content and hexokinase (HK) and pyruvate kinase (PK) activity. Twelve phenolic compounds were detected by UHPLC-Q-TOF-MS, α-glucosidase inhibitors (AGIs). This study suggests that PASs, as a by-product, has the potential to become a new raw material source for hypoglycemic functional food, providing an application prospect for commonly considered waste product.

Combined administration of Gegen Yinlian decoctio n and metformin ameliorates liver metabolic disorder in rats by regulating miR 195 5p/IRS1/PI3K/AKT axis

Purpose: To investigate the effect of Gegen Yinlian decoction (GYD), made from lobed kudzuvine root, baicalensis, huanglian and liquorice, and metformin on liver metabolism dysfunction in type 2 diabetes mellitus (T2DM) rats.
 Methods: Wistar rats were treated with high-fat diet and a single intraperitoneal streptozotocin (STZ) injection to establish T2DM model. Thereafter, the T2DM-rats were treated with GYD and metformin (GYD/metformin). Serum triglyceride (TG), TC, LDL, and HDL levels, as well as glucose and insulin tolerance of the rats were evaluated. Furthermore, the effects of GYD/metformin treatment on the expressions of the related genes were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunoblot assay.
 Results: After GYD-metformin treatment, the metabolic indicators in T2DM rats significantly improved (p < 0.05). However, there was decrease in t liver miR-1955p expressions. Silencing miR-195-5p improved glucose consumption and triglyceride levels of palmitate (PA)-induced pathological state in ALM12 cells (p < 0.05). Furthermore, miR-195-5p targeted insulin receptor substrate 1 (IRS1). Suppression of IRS1 partly annulled the impact of miR-195-5p silencing on ALM12 cell model. Moreover, IRS1 downregulation significantly mitigated the impact of the silencing on PI3K/AKT pathway activity (p < 0.05).
 Conclusion: Combination of GYD with metformin ameliorated dysfunctional liver metabolism via control of miR-195-5p/IRS1/PI3K/AKT axis.

Screening and identification of a novel antidiabetic peptide from collagen hydrolysates of Chinese giant salamander skin: network pharmacology, inhibition kinetics and protection of IR-HepG2 cells.

In this study, a novel peptide GPPGPA was screened from the collagen hydrolysates of Chinese giant salamander (Andrias davidianus) skin, and its anti-diabetes mechanism was predicted by network pharmacology, and an inhibitory effect on α-glycosidase and protective effect on IR (insulin resistance) and oxidative stress of IR-HepG2 cells were detected. Through network pharmacology screening, GPPGPA was found to have good drug-like properties, and 103 targets of GPPGPA overlap with T2DM targets. These targets were mainly enriched in the PI3K-Akt signaling pathway associated with T2DM, the AGE-RAGE signaling pathway in diabetic complications, the TNF signaling pathway, insulin resistance and so on. The core targets were identified as AKT1, MAPK8, MAPK10 and JUN by constructing a "peptide-target-pathway" network. The molecular docking results showed that GPPGPA was well bound to the core targets. These results suggested that GPPGPA had the potential to reduce T2DM. Further in vitro experiments showed that GPPGPA as a competitive inhibitor could effectively inhibit the activity of α-glucosidase. The results of the IR-HepG2 cell model experiments showed that GPPGPA was not toxic to HepG2 cells, and could reduce IR of HepG2 cells induced by high-glucose and high-insulin, improve glucose consumption, increase the activity of superoxide dismutase (SOD), and reduce the content of malondialdehyde (MDA). The above results suggested that GPPGPA could improve T2DM by reducing insulin resistance through a multi-target and multi-pathway mechanism. GPPGPA could be developed and utilized as a novel hyperglycemic inhibitor in functional food.

Transcriptional and Metabolic Profiling of Nicotinamide-Enhanced Natural Killer (NAM-NK) Cells (GDA-201)

Adoptive transfer of NK cells is a promising immunotherapeutic modality, however limited NK cell persistence and proliferation in vivo have historically been barriers to clinical success. Nicotinamide (NAM), an allosteric inhibitor of NAD-dependent enzymes, has been shown to preserve cell function and prevent differentiation in ex vivo culture of NK (NAM-NK) and other cells. Clinical responses were observed in a Phase 1 trial of NAM-NK (GDA-201) in patients with refractory non-Hodgkin lymphoma (Bachanova, et. al., Blood 134:777, 2019). We now use transcriptional and metabolic profiling to characterize the mechanisms underlying the activity of NAM-NK.CD3 negative lymphocytes obtained from healthy donors were cultured for 14 days with IL-15 in the presence or absence of NAM (7 mM). Next generation sequencing (NGS), liquid chromatography-mass spectrometry (LC-MS)-based metabolite quantification, and glycolytic/mitochondrial respiration measurements were performed. Transcriptome and pathway enrichment analyses were performed with Ingenuity Pathway Analysis software. Extracted cellular and medium metabolites were analyzed on a Thermo Q-Exactive Plus mass spectrometer coupled with a Vanquish UHPLC system. Extracellular acidification (ECAR) and oxygen consumption rates (OCR) were quantified using a Seahorse Extracellular Flux Analyzer. Glycolysis/citric acid cycle (TCA) rates were measured using isotope-labelled glucose incorporation assays.Transcriptome analyses defined 1,204 differentially expressed (DE) genes in NAM-NK vs. control NK. Biological/functional enrichment and pathway analyses of DE-genes predicted upregulation of cell cycle, DNA replication (CDK4/CDKN2D, CyclinD/E, MAD2L), RNA transcription, translation (SMN1/2, ABCF1, EIF4B, RPL13, RPS6), protein synthesis (EIF2, PABPC1, SOS, 60S complex) mitochondrial energy metabolism (NDUFB8, ATP5G2/E, COX7B/C) migration, homing (CD62L, CD44, DNAM1), and cytokine/chemokine response (IL18R, CXCR3, CCR5, XCL1, SOCS3, LFA1) pathways, with concomitant downregulation of cell exhaustion, senescence (BATF1, FOXP1, STAT1, CD86, LGALS9, LAG3), apoptosis, necrosis (CASP1, MDM2, IKK3), stress response (CALR, HSP90, HSPH1), and lymphoid cellular maturation (IL-2Ra, CD40L, GATA3) pathways in NAM-NK. Metabolomic analyses showed a significant increase of intracellular NAD, NADH, NADP, NADPH, high-energy triphosphates (ATP, UTP, GTP) and overall energy charge ([ATP+0.5*ADP]/[ATP+ADP+AMP]) in NAM-NK. Cellular metabolic fitness analyses revealed increased basal and ATP-linked respiration, mitochondrial maximal respiratory capacity, and glycolytic capacity in NAM-NK compared to control NK. In addition, NAM increased the rate of glucose incorporation into TCA cycle intermediates (acetyl-CoA, succinyl-CoA), consistent with a more rapid glycolysis rate, increased TCA cycling, and improved glucose consumption efficiency.Taken together, results of transcriptome, metabolomic, mitochondrial respiration, and glycolytic rate analyses suggest that NAM pleiotropically modulates key cellular metabolic functions in ex vivo-expanded NK cells, resulting in increased response to cytokine stimulation and enhanced potency. NAM inhibits differentiation, cellular stress, and exhaustion pathways that are typically activated in culture. Moreover, NAM increases cellular metabolic fitness, energy charge, and efficiency of glucose consumption, potentially imparting a protective effect against oxidative stress in the tumor microenvironment. These data offer insight into the mechanism of improved persistence, proliferation, and cytotoxicity observed in in vivo and clinical studies of GDA-201. DisclosuresYackoubov: Gamida Cell: Current Employment. Pato: Gamida Cell: Current Employment. Rifman: Gamida Cell: Current Employment. Cohen: Gamida Cell: Current Employment. Hailu: Gamida Cell: Current Employment. Persi: Gamida Cell: Current Employment. Berhani-Zipori: Gamida Cell: Current Employment. Edri: Gamida Cell: Current Employment. Peled: Biokine Therapeutics Ltd: Current Employment; Gamida Cell: Research Funding. Cichocki: Gamida Cell: Research Funding; Fate Therapeutics, Inc: Patents & Royalties, Research Funding. Rabinowitz: Gamida Cell: Research Funding. Lodie: Gamida Cell: Current holder of stock options in a privately-held company, Ended employment in the past 24 months. Adams: Gamida Cell: Current Employment. Simantov: Gamida Cell: Current Employment. Geffen: Gamida Cell: Current Employment.

Discovery of benzimidazole derivatives as potent and selective aldehyde dehydrogenase 1A1 (ALDH1A1) inhibitors with glucose consumption improving activity

Aldehyde dehydrogenase 1A1 (ALDH1A1) plays vital physiological and toxicological functions in many areas, such as CNS, inflammation, metabolic disorders, and cancers. Overexpression of ALDH1A1 has been disclosed to play an important role in obesity, diabetes and other diseases, indicating the potential need for the identification and development of small molecule ALDH1A1 inhibitors. Herein, a series of benzimidazole derivatives was designed, synthesized and evaluated. Among them, compounds 21, 27, 29, 61 and 65 exhibited excellent inhibitory activity against ALDH1A1 with IC50 values in the low micromolar range and high selectivity over ALDH1A2, ALDH1A3, ALDH2 and ALDH3A1. Moreover, an in vitro study demonstrated that all five compounds effectively improved glucose consumption in HepG2 cells, of which, 61 and 65 at 10 µM produced nearly equal glucose consumption with positive control Metformin (Met) at 1 mM. Furthermore, 61 and 65 showed desirable metabolic stability in human liver microsomes. All these results suggest that 61 and 65 are suitable for further studies.

Structural characteristics of a hypoglycemic polysaccharide from Fructus Corni

PFC-3 is a homogeneous polysaccharide extracted from the dried pulps of Fructus Corni with a molecular weight of 40.3 kDa. The crude polysaccharide was obtained and further purified by DEAE-Sephadex A-25 and Sephadex G-100 columns to investigate its structure and glycemic effect. The monosaccharides in the PFC-3, determined by high-performance liquid chromatography, consisted of glucose (Glc), xylose (Xyl), and galactose (Gal) with a mass molar ratio of 2.35:12.49:1.00. The methylation analysis combined with 1D (1H and 13C), and 2D NMR (1H–1H COSY, HSQC, and HMBC) further demonstrated that PFC-3 was mainly composed of 1,3-α-D-Xylp, 1,6-α-D-Galp, 1,2-α-D-Glcp, and T-α-D-Galp, and contained a backbone fragment of →6)-α-D-Galp-(1 → 2)-α-D-Glcp-(1 → 3)-α-D-Xylp-(1 → . The hypoglycemic effect of PFC-3 in vitro was evaluated by glucose uptake and consumption assays, and the results showed that PFC-3 concentration-dependently enhanced glucose uptake and significantly improved glucose consumption in insulin-resistant HepG2 cells. Furthermore, PFC-3 significantly reduced fasting blood glucose level, glycosylated hemoglobin level, amylase activity, ameliorate lipid metabolism, and hepatic lesions in streptozotocin-induced diabetic rats. Our research provided insights into the hypoglycemic activities of PFC-3.

The glucuronide metabolites of kaempferol and quercetin, targeting to the AKT PH domain, activate AKT/GSK3β signaling pathway and improve glucose metabolism

Abstract Kaempferol and quercetin, as dietary flavonoids, are beneficial to diabetes risk reduction. Clarifying the mechanism of dietary flavonoids in improving glucose metabolism may meet the functional and nutritional requirements. In this paper, PCA revealed two glucuronides, kaempferol-3-glucuronide (K-3-G) and quercetin-3-glucuronide (Q-3-G), which were the primary metabolites. Target prediction, enzymatic activity, and AKT plasma translocation assays indicated that K-3-G and Q-3-G could target the AKT PH domain. Fluorescence spectrometry was used to measure the dissociation constants of K-3-G (KD = 27.6 μM) and Q-3-G (KD = 10.8 μM). Molecular docking provided interaction insights for K-3-G and Q-3-G with the AKT PH domain. K-3-G and Q-3-G activated the AKT/GSK3β phosphorylation, thereby improving glucose consumption. Our study demonstrates that the glucuronide metabolites of flavonoids directly target the AKT PH domain and activate AKT/GSK3β signal pathway, thereby improving glucose metabolism.

Structural characterization of polysaccharide from Centipeda minima and its hypoglycemic activity through alleviating insulin resistance of hepatic HepG2 cells

• A homogeneous heteropolysaccharide was isolated and purified from Centipeda minima . • The backbone chain of CMP-2B were 3,6)-α-D-Man p -(1 and 4,6)-β-D-Gal p -(1. • CMP-2B possessed preferable antioxidant and α-glucosidase inhibitory activity. • CMP-2B could improve insulin resistant in IR-HepG2 cells. A novel polysaccharide (CMP-2B) was purified from Centipeda minima by anion-exchange and gel-permeation chromatography. Structural characterization revealed that CMP-2B was a homogeneous heteropolysaccharide with an average molecular weight of 113.193 KDa. CMP-2B was made up of mannose, galacturonic acid, galactose and arabinose with a molar ratio of 0.27:0.12:0.42:0.17. The backbone chain of CMP-2B consisted of 3,6)-α-D-Man p -(1 and 4,6)-β-D-Gal p -(1, and the branches were comprised of 4)-α-D-Gal p A-(1, 6)-β-D-Gal p -(1, T-α-L-Ara f -(1 and 3)-α-L-Ara f -(1. Besides, the results of antioxidant assays revealed that CMP-2B possessed signifcant free radical scavenging ability. Moreover, the results of hypoglycemic assays showed that CMP-2B displayed α-glucosidase inhibitory activities, and could significantly improve glucose consumption, glycogen synthesis and the activity of pyruvate kinase of insulin-resistance HepG2 cells. Overall, these results suggested that CMP-2B could be a new source for functional foods.

Hengshun Aromatic Vinegar Improves Glycolipid Metabolism in Type 2 Diabetes Mellitus via Regulating PGC-1α/PGC-1β Pathway.

Hengshun aromatic vinegar (HSAV), produced by typical solid-state or liquid-state fermentation techniques, is consumed worldwide as a food condiment. HSAV shows multiple bioactivities, but its activity in type 2 diabetes mellitus (T2DM) and possible mechanisms have not been reported. In this study, the effects of HSAV against T2DM were evaluated in insulin-induced HepG2 cells and high-fat diet (HFD) and streptozotocin (STZ) induced T2DM rats. Then, the mechanisms of HSAV against T2DM were explored by Real-time PCR, Western blot, immunofluorescence assays, siRNA transfection and gene overexpression experiments. Results indicated that HSAV significantly improved glucose consumption and reduced triglycerides (TG) contents in metabolic disordered HepG2 cells. Meanwhile, HSAV obviously alleviated general status, liver and kidney functions of T2DM rats, and decreased hyperglycemia and hyperlipidemia, improved insulin resistance, and reduced lipid accumulation in liver. Mechanism studies indicated that HSAV markedly down-regulated the expression of proliferator-activated receptor γ coactivator-1α (PGC-1α), then regulated peroxisome proliferators-activated receptor α (PPAR-α)/protein kinase B (AKT) signal pathway mediated gluconeogenesis and glycogen synthesis. Meanwhile, HSAV significantly up-regulated proliferator-activated receptor γ coactivator-1β (PGC-1β), and subsequently decreased sterol regulatory element binding protein-1c (SREBP-1c) pathway mediated lipogenesis. In conclusion, HSAV showed potent anti-T2DM activity in ameliorating dysfunction of glycolipid metabolism through regulating PGC-1α/PGC-1β pathway, which has a certain application prospect as an effective diet supplement for T2DM therapy in the future.

Humanin Alleviates Insulin Resistance in Polycystic Ovary Syndrome: A Human and Rat Model-Based Study.

Polycystic ovary syndrome (PCOS), the most common endocrine disorder in women of reproductive age, is characterized by hyperandrogenism and insulin resistance (IR); however, the pathogenesis of local ovarian IR in PCOS remains largely unclear. Humanin, a mitochondria-derived peptide, has been reported to be associated with IR. Our previous study confirmed that humanin is expressed in multiple cell types in the ovary and is present in follicular fluid. However, it remains unknown whether humanin participates in the pathogenesis of local ovarian IR or whether humanin supplementation can improve IR in PCOS patients. In this study, we compared humanin concentrations in follicular fluid from PCOS patients with and without IR. We further investigated the effect of humanin analogue (HNG) supplementation on IR in a rat model of dehydroepiandrosterone-induced PCOS. Humanin concentrations in the follicular fluid were found to be significantly lower in PCOS patients with IR than in those without IR. HNG supplementation attenuated both the increases in the levels of fasting plasma glucose and fasting insulin in rats with PCOS and the decreases in phosphorylation of IRS1, PI3K, AKT, and GLUT4 proteins in the granulosa cells of these rats. Combined supplementation with HNG and insulin significantly improved glucose consumption in normal and humanin-siRNA-transfected COV434 cells. In conclusion, downregulated humanin in the ovaries may be involved in the pathogenesis of IR in PCOS, and exogenous supplementation with HNG improved local ovarian IR through modulation of the IRS1/PI3K/Akt signaling pathway in a rat model. This finding supports the potential future use of HNG as a therapeutic drug for PCOS.

Tetrahydrocannabinol-Rich Extracts From Cannabis Sativa L. Improve Glucose Consumption and Modulate Metabolic Complications Linked to Neurodegenerative Diseases in Isolated Rat Brains.

Reduced brain glucose consumption arising from impaired glucose uptake and utilization has been linked to the pathogenesis and complications of neurodegenerative diseases. The ability of Cannabis sativa L. tetrahydrocannabinol (THC)-rich extracts to stimulate brain glucose uptake and utilization as well as its modulatory effect on gluconeogenesis, antioxidative, purinergic and cholinergic activities were investigated in isolated rats’ brains. C. sativa leaves were sequentially extracted to yield the hexane and dichloromethane extracts. The extracts were incubated at 37°C with freshly harvested brains in the presence of glucose for 2 h. The control consisted of incubation without the extracts, while brains without the extracts and glucose served as the normal control. Metformin was used as the standard drug. C. sativa extracts caused a significant (p < 0.05) increase in brain glucose uptake, with concomitant elevation of glutathione level, superoxide dismutase, catalase, and ecto-nucleoside triphosphate diphosphohydrolase activities compared to the controls. Incubation with C. sativa extracts also led to depletion in malondialdehyde and nitric oxide levels, acetylcholinesterase, butyrylcholinesterase, glucose 6-phosphatase and fructose-1,6-biphosphatase activities. GC-MS analysis of the extracts revealed the presence of THC. In silico analysis predicted THC to be permeable across the blood-brain-barrier. THC was also predicted to have an oral LD50 and toxicity class values of 482 mg/kg and 4 respectively. These results indicate that C. sativa improves glucose consumption with concomitant suppression of oxidative stress and cholinergic dysfunction, and modulation of purinergic and gluconeogenic activities in brain tissues

Design, synthesis of 1,3-dimethylpyrimidine-2,4-diones as potent and selective aldehyde dehydrogenase 1A1 (ALDH1A1) inhibitors with glucose consumption improving activity

LDH1A1, one of 19 NAD(P)+-dependent aldehyde dehydrogenases, participates in multiple metabolic pathways and has been indicated to play an important role in obesity and diabetes. In this study, a series of 1,3-dimethylpyrimidine-2,4-diones were designed, synthesized and evaluated as novel selective aldehyde dehydrogenase 1A1 inhibitors. Among them, compounds 46, 50, 53, 56 and 57 exhibited excellent inhibitory activity against ALDH1A1 with IC50 values in the low nanomolar range and high selectivity over ALDH1A2, ALDH1A3, ALDH2 and ALDH3A1. Furthermore, in vitro study demonstrated that compound 57 effectively improved glucose consumption in HepG2 cells compared to compound 1 (CM026).

Enhanced biohydrogen production from cotton stalk hydrolysate of Enterobacter cloacae WL1318 by overexpression of the formate hydrogen lyase activator gene

BackgroundBiohydrogen production from lignocellulose has become an important hydrogen production method due to its diversity, renewability, and cheapness. Overexpression of the formate hydrogen lyase activator (fhlA) gene is a promising tactic for enhancement of hydrogen production in facultative anaerobic Enterobacter. As a species of Enterobacter, Enterobacter cloacae was reported as a highly efficient hydrogen-producing bacterium. However, little work has been reported in terms of cloning and expressing the fhlA gene in E. cloacae for lignocellulose-based hydrogen production.ResultsIn this study, the formate hydrogen lyase activator (fhlA) gene was cloned and overexpressed in Enterobacter cloacae WL1318. We found that the recombinant strain significantly enhanced cumulative hydrogen production by 188% following fermentation of cotton stalk hydrolysate for 24 h, and maintained improved production above 30% throughout the fermentation process compared to the wild strain. Accordingly, overexpression of the fhlA gene resulted in an enhanced hydrogen production potential (P) and maximum hydrogen production rate (Rm), as well as a shortened lag phase time (λ) for the recombinant strain. Additionally, the recombinant strain also displayed improved glucose (12%) and xylose (3.4%) consumption and hydrogen yield Y(H2/S) (37.0%) compared to the wild strain. Moreover, the metabolites and specific enzyme profiles demonstrated that reduced flux in the competitive branch, including succinic, acetic, and lactic acids, and ethanol generation, coupled with increased flux in the pyruvate node and formate splitting branch, benefited hydrogen synthesis.ConclusionsThe results conclusively prove that overexpression of fhlA gene in E. cloacae WL1318 can effectively enhance the hydrogen production from cotton stalk hydrolysate, and reduce the metabolic flux in the competitive branch. It is the first attempt to engineer the fhlA gene in the hydrogen-producing bacterium E. cloacae. This work provides a highly efficient engineered bacterium for biohydrogen production from fermentation of lignocellulosic hydrolysate in the future.

Discovery and anti-diabetic effects of novel isoxazole based flavonoid derivatives

3-O-[(E)-4-(4-cyanophenyl)-2-oxobut-3-en-1-yl] kaempferol is a novel lead compound to discover anti-diabetic and anti-obesity drugs. The present study reported the scaffold hopping of the lead compound to obtain a new isoxazole derivative, C45, which has improved glucose consumption at the nanomolar level (EC50 = 0.8 nM) in insulin resistant (IR) HepG2 cells. Western blotting showed that C45 markedly enhanced the phosphorylation of AMPK (AMP-activated protein kinase) and reduced the levels of the gluconeogenesis key enzymes PEPCK (phosphoenolpyruvate carboxykinase) and G6Pase (glucose 6-phosphatase) in HepG2 cells. The potential molecular mechanism of C45 may be activation of the AMPK/PEPCK/G6Pase pathways. We concluded that C45 might be a valuable candidate to discover anti-diabetic drugs.

Flavonoid derivatives synthesis and anti-diabetic activities

In high fat diet-induced obese mice, the flavonoid derivative of tiliroside, Fla-CN, has antihyperglycemic effects, can improve insulin sensitivity, ameliorate metabolic lipid disorders, and benefits certain disorders characterized by insulin resistance. Fla-CN is a novel lead compound to discovery anti-diabetic and anti-obesity drugs. The present study reported the optimization of Fla-CN to obtain a new derivative, 10b, which has improved glucose consumption at the nanomolar level (EC50 = 0.3 nM) in insulin resistant (IR) HepG2 cells. 10b also increased the glycogen content and glucose uptake, and concurrently inhibited gluconeogenesis in HepG2 cells. Western blotting showed that 10b markedly enhanced the phosphorylation of AMPK (AMP-activated protein kinase) and AS160 (protein kinase B substrate of 160 kDa) and reduced the levels of the gluconeogenesis key enzymes PEPCK (phosphoenolpyruvate carboxykinase) and G6P (glucose 6-phosphatase) in HepG2 cells. The potential molecular mechanism of 10b may be activation of the AMPK/AS160 and AMPK/PEPCK/G6P pathways. We concluded that 10b might be a valuable candidate to discover anti-diabetic drugs.