Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 2 | Activating effects of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of PD173074 (FGFR1 inhibitor, 50 nmol L−1) or palmitic acid (PA, 500 µmol L−1), on FGF21 signaling in HepG2 human hepatocytes. Coffee by-product’s bioactive compounds interacted in silico with the subunits of the FGF21 receptor (FGFR) (A), exhibiting different binding (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: In Silico, Binding Assay

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 4 | Role of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (PA, 500 µmol L−1) on lipo/cytotoxicity and inflammation markers in HepG2 human hepatocytes. Coffee by-products’ bioactive compounds preserved cell viability (A) and diminished lactate dehydrogenase (LDH) release (B). PA-triggered tumor necrosis factor α (TNF-α) (C), interleukin (IL)-6 (D), and IL-1β (E) release and nitric oxide synthase (NOS) activity (F) were reduced. Hierarchical cluster analysis and heat map [from the lowest (■red) to the highest (■green) value for each parameter] (G) and an integrative diagram illustrating the effects of the bioactive compounds from coffee by-products on lipo/cytotoxicity and inflammation (H). The results are expressed as mean ± SD (n = 3). Bars with different letters significantly (p < 0.05) differ according to ANOVA and Tukey’s multiple range test. NT, non-treated cells; CAF, caffeine; CGA, chlorogenic acid; CA, caffeic acid; PCA, protocatechuic acid; GA, gallic acid; KMP, kaempferol; FGF21, fibroblast growth factor 21.

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: Activity Assay

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 5 | Protective effects of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (PA, 500 µmol L−1) against oxidative stress in HepG2 human hepatocytes. Coffee by-products’ bioactive compounds diminished the production of reactive oxygen species (ROS) (A) and mitochondrial O•− 2 (B), preserving the mitochondrial membrane potential (19m) (C) in PA-challenged HepG2 cells. The enzymatic activity NADPH oxidase (NOX) (D), superoxide dismutase (SOD) (E), and catalase (F) activities, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) phosphorylation (G) were regulated, thereby diminishing oxidative stress. Hierarchical cluster analysis and heat map [from the lowest (■red) to the highest (■green) value for each parameter] (H) and an integrative diagram illustrating the effects of the bioactive compounds from coffee by-products on oxidative stress (I). The results are expressed as mean ± SD (n = 3). Bars with different letters significantly (p < 0.05) differ according to ANOVA and Tukey’s multiple range test. NT, non-treated cells; CAF, caffeine; CGA, chlorogenic acid; CA, caffeic acid; PCA, protocatechuic acid; GA, gallic acid; KMP, kaempferol; FGF21, fibroblast growth factor 21.

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: Preserving, Membrane, Activity Assay, Derivative Assay, Phospho-proteomics

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 6 | Regulative role of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (500 µmol L−1) on the mitochondrial bioenergetics of HepG2 human hepatocytes. Coffee (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques:

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 7 | Modulatory effects of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (500 µmol L−1) on lipid metabolism in HepG2 human hepatocytes. Palmitic acid-treated (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques:

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 8 | Regulatory role of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (500 µmol L−1) on glucose metabolism in HepG2 human hepatocytes. Hepatocytes exhibited a modulation on glucose uptake (A), glucokinase (GK) activity (B), glucose production (C), phosphoenolpyruvate carboxykinase (PEPCK) activity (D), insulin receptor (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: Activity Assay

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 9 | Integrative hierarchical cluster analysis and heat map (from the lowest (■red) to the highest (■green) value for each parameter) unifying all parameters demonstrates that chlorogenic and protocatechuic acids are the bioactive compounds exhibiting the highest NAFLD-protecting effects (A). Diagram illustrating the molecular mechanisms from the effects of the bioactive compounds in coffee by-products on hepatic FGF21 signaling, oxidative stress, mitochondrial bioenergetics, and energy metabolism (B). NT, non-treated cells; PA, palmitic acid; CAF, caffeine; CGA, chlorogenic acid; CA, caffeic acid; PCA, protocatechuic acid; GA, gallic acid; KMP, kaempferol; FGF21, fibroblast growth factor 21.

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: GLN’s induction of FGF21 expression in the hippocampus, cortex, and striatum. After two weeks of GLN injection, the protein levels of FGF21 were measured in the hippocampus, cortex, and striatum of ND ( A ) or HFD mice ( B ) using Western blotting with β-actin as an internal control. The results represent the means ± S.E.M. ( n = 8). * p < 0.05 and *** p < 0.001 compared with the GLN 0 mg/mouse group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Injection, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: GLN-induced expression of FGF21 in hippocampal and striatal cells. HT22 hippocampal ( A ) and STHdh Q7/Q7 striatal cells ( B ) were treated with GLN (0, 1.25, 2.5, 5, 10, 20 mM) for 6 h for mRNA determination or 24 h for protein measurement. FGF21 mRNA was quantified using real-time PCR with GAPDH as an internal control, while FGF21 protein was examined through Western blotting with β-actin as an internal control. FGF21 production in the cultured medium was assessed with ELISA. The results represent the means ± S.E.M. ( n = 4). * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the GLN 0 mM group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: Attenuation of GLN-enhanced learning and memory functions and FGF21 production through FGFR1 inhibition in the hippocampus, hippocampal cells, and striatal cells. The ND-fed mice were administrated GLN (0, 30 mg/mouse) alone or in combination with PD173074 (2 mg/kg) through IP injection for 2 weeks, followed by the NORT ( A ). The FGF21 protein levels in the hippocampus ( B ) and in HT22 or STHdh Q7/Q7 cells treated with GLN (20 mM) alone or in combination with PD173074 (0.025, 0.1, 1 μM) for 24 h ( C ) were determined through Western blotting or ELISA. The results represent the means ± S.E.M. from 6 animals ( A ) or 4 animals ( B ) in each treatment group or from 4 independent experiments ( C ). * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the GLN 0 mg/mouse group or the GLN 0 mM group. # p < 0.05 and ### p < 0.001 compared with the GLN 30 mg/mouse or the GLN 20 mM only group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Inhibition, Injection, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: Illustration of the potential involvement of signaling molecules in GLN-mediated FGF21 production in hippocampal and striatal cells. ( A ) HT22 cells or ( B ) STHdh Q7/Q7 cells were pretreated with specific inhibitors targeting NF-κB (Parthenolide, 5 µM), MAPK p38 (SB203580, 20 µM), JNK (SP600125, 20 µM), ERK (PD98059, 20 µM), Akt (LY294002, 10 μM), PKA (H89, 5 µM), PKC (Bisindolylmaleimide, BIMI, 5 µM), PPARα (GW6471, 10 µM), or PPARγ (GW9662, 10 µM) for 1 h before the addition of GLN (20 mM) for an additional 24 h. The FGF21 concentration in the cultured medium was quantified using ELISA. The results represent the means ± S.E.M. ( n = 4). * p < 0.05 and *** p < 0.001 compared with the 0 mM group. # p < 0.05 and ### p < 0.001 compared with the GLN 20 mM group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: Demonstration of the further induction of FGF21 through overexpression of NF-κB p65 in HT22 cells. HT22 cells were transfected with EGFP-p65 or EGFP plasmid and treated with GLN (20 mM) for 24 h. Western blotting was performed to detect the proteins for FGF21, GFP, EGFP-RelA, and p65 using β-actin as an internal control. The results represent the means ± S.E.M. ( n = 3). * p < 0.05 and *** p < 0.001 compared with the 0 mM group. && p < 0.01 between the EGFP and EGFP-RelA groups.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Over Expression, Transfection, Plasmid Preparation, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Construction, purification, and characterization of FGF21-164. ( A ) FGF21-164 sequence and FGF21 sequence. In the FGF21-164 sequence, red marked the mutated amino acid and blue represents the TR acid sequence from hCD164. In the FGF21 sequence, the red-labeled amino acid represents the amino acid sequence whose n-terminal is deleted in FGF21-164. ( B ) Subcloned cell culture supernatants expressing FGF21-164 and FGF21 were analyzed using Western blotting (Lane M: molecular weight marker. Lane 1~7: FGF21-164 clones 1, 2, 3, A4, B2, F3, and H4. Line 8: FGF21 clone2). ( C ) The purified FGF21-164 and FGF21 were assayed using SDS-PAGE and Western blotting (Lane M: molecular weight marker. Lane 1: purified FGF21-164. Line 2: purified FGF21). ( D ) Purity and hydrodynamic radius of FGF21-164 were analyzed using SEC-HPLC. ( E ) MALDI-TOF mass-spectrometry analysis of FGF21-164. ([M + H] + denotes the singly charged ionic forms, and [M + 2H] 2+ denotes doubly charged ionic forms). ( F ) Circular Dichroism (CD) spectra of FGF21-164 and FGF21.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Purification, Sequencing, Labeling, Cell Culture, Expressing, Western Blot, Molecular Weight, Marker, Clone Assay, SDS Page, Mass Spectrometry, Circular Dichroism

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Glycan forms analysis of the protein FGF21-164. ( A ) O-linked glycan forms of FGF21-164. ( B ) N-linked glycan forms of FGF21-164.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of FGF21-164 in adipocytes induced by 3T3-L1. ( A ) The process of inducing 3T3-L1’s differentiation into adipocytes. ( B ) Reduction in glucose in medium stimulated by FGF21-164 in adipocytes induced by 3T3-L1. Data are means ± SEM (n = 6). * p < 0.05, *** p < 0.001, and **** p < 0.0001 compared to 0 μg/mL FGF21-164. ( C ) Phospho-Erk1/2-specific bands (Thr202/Tyr204; 44–42 kDa) were detected in 3T3-L1-derived adipocytes after FGF21-164 stimulation. ( D ) Evaluation of lipid droplet accumulation in adipocytes induced by 3T3-L1 with or without FGF21-164. The scale bar represents 50 μm.Data are the means ± SEM (n = 6). * p < 0.05, **** p < 0.0001.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Pharmacokinetics of FGF21-164. FGF21-164 plasma levels in C57BL/6 mice as determined via targeted LC-MS after i.v. (tail vein) bolus injection of FGF21-164 protein.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Liquid Chromatography with Mass Spectroscopy, Injection

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: The results of an oral glucose tolerance test applied to ob / ob mice after a single FGF21-164 treatment. ( A ) Blood glucose levels were measured at 0, 2, 3, 4, 5, 6, 7, and 8 h after the administration of FGF21-164. Data are the means ± SEM (n = 4–5). * p < 0.05 and ** p < 0.01 compared with the model. ( B ) The decrease in blood glucose levels post-administration relative to the baseline measurement at 0 h. Data are the means ± SEM (n = 4–5). * p < 0.05 compared with the model.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of a single administration of FGF21-164 on ob / ob mice. ( A , D , G ) Fasting glucose levels. Blood glucose levels were measured at 7, 21, and 28 days after a single dose of FGF21-164 was administered. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, ** p < 0.01, and **** p < 0.0001. ( B , E , H ) The OGTT at 7, 21, and 28 days. After oral glucose administration, blood glucose levels were measured. Data are the means ± SEM (n = 4–5). * p < 0.05, *** p < 0.001, and **** p < 0.0001 at 6 mg·kg −1 vs. model. # p < 0.05, ## p < 0.01 and #### p < 0.0001 at 12 mg·kg −1 vs. model. ( C , F , I ) The glucose AUC during the OGTT process. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, *** p < 0.001. ( J ) Body weight measured at 0, 7, 21, and 28 days. Data are the means ± SEM (n = 4–5). ns: no significance.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of repeated administration of FGF21-164. ( A ) Body weight changes. The DIO mice were treated with PBS or FGF21-164 once every 2 days. Data are the means ± SEM (n = 5–7). **** p < 0.0001 vs. Ctrl. #### p < 0.0001 for FGF21-164 vs. the model. ( B ) The liver sections were stained with Oil-red-O. The scale bar represents 100 μm. DIO mice showed many red lipid droplets in hepatic tissue (indicated by the black arrow), while lipid droplet levels were lower after the FGF21-164 treatment.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Staining