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: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Concentration-dependence of BrdU incorporation in A2B5+ OPCs cultured with adult mice serum (n = 6). (B) BrdU incorporation in A2B5+ OPCs 1 day after stimulation with adult mouse serum heated or pretreated with the indicated reagents (n = 6). (C) BrdU incorporation in OPCs after serum stimulation with PD173074 (10 nM), an inhibitor of FGFR (n = 4). (D) BrdU incorporation in OPCs after serum stimulation with NF449 (10 μM), an inhibitor of FGFR3 (n = 4). (E) BrdU incorporation in mouse OPCs with FGFR and β-klotho knockdown after serum stimulation (n = 7). (F) BrdU incorporation in OPCs after stimulation with recombinant FGF15, FGF21, and FGF23 (n = 4). (G) BrdU incorporation in OPCs after serum stimulation with neutralizing antibody against FGF21 (n = 5), determined by Student’s t test or by ANOVA with Tukey’s post hoc test or Dunnett’s test. Error bars represent SEM. *P < 0.05, **P < 0.01.

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Concentration Assay, BrdU Incorporation Assay, Cell Culture, Recombinant

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Quantitations of Fgf21 mRNA (left) and FGF21 protein (right) in individual organs of intact mice (n = 9); **P < 0.01. (B) Representative images of FGF21-immunolabeled pancreas of intact mice (n = 3). (C) Double IHC staining for FGF21 with the indicated cell markers in the pancreas of adult mice (n = 3). (D) BrdU incorporation in mouse OPCs after stimulation with serum from mice with FGF21 knockdown in the pancreas (n = 4); *P < 0.05, **P < 0.01, as determined by ANOVA with Tukey’s post hoc test. Error bars represent SEM. Scale bars: 50 μm (B); 10 μm (C).

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Immunolabeling, Immunohistochemistry, BrdU Incorporation Assay

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Quantitation of FGF21 protein in the spinal cord 1 day and 3 days after LPC injection (n = 5 for control, 5 for d1, 4 for d3). (B) Representative images of spinal cord sections double-labeled for PDGFRα and Ki67. Sections were obtained from FGF21-KO mice and control littermates 7 days after LPC injection. Graph shows quantitations as indicated in the images (n = 5). Arrows indicate Ki67+ cells colabeled with PDGFRα; arrowheads indicate Ki67+ cells not labeled with PDGFRα. (C) Representative images of spinal cord sections labeled for MBP. Sections were obtained from mouse spinal cord 14 days after LPC injection. Graph shows quantitations as indicated in the images (n = 5). (D) Representative immunoelectron microscopy images of myelin in the spinal cord. Sections were obtained from FGF21-KO mice and control littermates 14 days after LPC injection. Graphs show quantitations of g-ratio indicated in the images (n = 3). (E) Motor function was assessed by ladder-walk test (n = 11 for control littermates, 9 for FGF21-KO mice). (F) Representative images of spinal cord sections labeled for MBP. Graph shows quantitations as indicated in the images (n = 5 for control littermates + vehicle, 5 for FGF21-KO mice + vehicle, 4 for FGF21-KO mice + FGF21). (G) Representative images of spinal cord sections labeled for MBP. Graph shows quantitations as indicated in the images (n = 4); **P < 0.01 as determined by Student’s t test or by ANOVA with Tukey’s post hoc test or Dunnett’s test. *P < 0.05, **P < 0.01. Error bars represent SEM. Scale bars: 50 μm (B); 200 μm (C, F, and G); 2 μm (D).

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Quantitation Assay, Injection, Labeling, Immuno-Electron Microscopy

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Representative images of β-klotho expression in the mouse spinal cord 3 days after LPC injection. (B) Representative images of spinal cord sections double-labeled for PDGFRα and Ki67. Graph shows quantitations as indicated in images (n = 3 for control, 3 for CKO); *P < 0.05. Arrows indicate Ki67+ cells colabeled with PDGFRα. (C) Representative images of spinal cord sections labeled for MBP. Graph shows quantitations as indicated in the images (n = 4 each); **P < 0.01. (D) Representative images of brain sections double-labeled for PDGFRα and Ki67 in the mouse cortex, 7 days after traumatic brain injury. FGF21 was administered i.c.v. 24 hours after LPC injection (n = 4); *P < 0.05. Arrows indicate Ki67+ cells colabeled with PDGFRα; arrowheads indicate Ki67+ cells not labeled with PDGFRα. (E) Representative images of brain sections labeled for MBP in the mouse cortex, 14 days after traumatic brain injury. Graph shows quantitations as indicated in the images (n = 6 for vehicle, n = 4 for FGF21); *P < 0.05. (F) Representative immunoelectron microscopy images of myelin in the mouse cortex, 14 days after traumatic brain injury. Graphs show quantitations of g-ratio indicated in the images (n = 4); **P < 0.01 as determined by Student’s t test. Error bars represent SEM. Scale bars: 20 μm (A); 50 μm (B and D); 200 μm (C and E); 2 μm (F).

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Expressing, Injection, Labeling, Immuno-Electron Microscopy

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Representative image of β-klotho expression in an autopsied sample from healthy patient and a patient with multiple sclerosis. Graphs show quantitations as indicated in the images (n = 4 for healthy patients, 3 for multiple sclerosis patients); **P < 0.01. (B) BrdU incorporation in human OPCs after stimulation with recombinant FGF21 (n = 6 for control, 4 for FGF21); *P < 0.05 as determined by Student’s t test. Error bars represent SEM. Scale bar: 20 μm.

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Expressing, BrdU Incorporation Assay, Recombinant

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Antibodies utilized in the current study.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques:

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Short‐term semaglutide treatment improves glucose tolerance and reduces body weight in HFD‐challenged mice. (a) Diagram shows the animal experimental design. (b) Body weight changes during last 7 days in the three indicated groups. (c) Blood glucose level and area under the curve (AUC) during IPGTT. (d) Fasting (overnight) blood glucose levels at the end of the experiment for indicated groups. (e–g) Fasting plasma leptin (e), adiponectin (f), and FGF21 (g) levels. (h–m) Fat pad weights including epididymal (h, eWAT) and inguinal (i, iWAT) white adipose tissue and brown adipose tissue (j, BAT). (k) eWAT weight to body weight ratio. (l) iWAT weight to body weight ratio. (m) BAT weight to body weight ratio. Sema, semaglutide. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Clinical Proteomics

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment restores HFD‐induced attenuation on ERK phosphorylation to hFGF21 treatment in hepatocytes. (a) Western blotting show expression levels of indicated protein in MPH isolated from LFD‐fed mice after 1 h hFGF21 treatment with indicated dose. (b, c) Densitometric analyses for pERK (p44, Thr202) and pERK (p42, Tyr204) with indicated treatment. (d) Western blotting show expression levels of indicated protein in MPH isolated from HFD‐fed mice after 1 h hFGF21 treatment with indicated dose. (e, f) Densitometric analyses for pERK (p44, Thr202) and pERK (p42, Tyr204) with indicated treatment. (g) Western blotting show expression levels of indicated protein in MPH isolated from semaglutide‐treated mice after 1 h hFGF21 treatment with indicated dose. (h, i) Densitometric analyses for pERK (p44, Thr202) and pERK (p42, Tyr204) with indicated treatment. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Phospho-proteomics, Western Blot, Expressing, Isolation

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment restores the stimulatory effects of hFGF21 on its downstream target gene expressions in MPH. (a) qRT‐PCR show effect of indicated dose of hFGF21 treatment (4 h) on expression of cFos in MPH isolated from LFD‐fed, HFD‐fed, and semaglutide‐treated mice. (b) qRT‐PCR show effect of indicated dose of hFGF21 treatment (4 h) on expression of Egr1 in MPH isolated from LFD‐fed, HFD‐fed, and semaglutide‐treated mice. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Quantitative RT-PCR, Expressing, Isolation

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment improves FGF21 sensitivity and attenuates the effect of HFD feeding on hepatic gene expression. (a) Western blotting show expression levels of FGF21 in the liver of three indicated groups. (b–d) qRT‐PCR shows the comparison of expression levels on Fgf21 (b) and genes that encode its receptor, Fgfr1 (c) and co‐receptor Klb (d) in the liver of three indicated groups. (e) qRT‐PCR shows the comparison of expression levels of lipogenic and fatty acid oxidation genes in the liver of three indicated groups. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Gene Expression, Western Blot, Expressing, Quantitative RT-PCR, Comparison

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment recovers HFD‐induced alteration in a battery of adipose‐specific genes. (a–c) qRT‐PCR shows the comparison of expression levels on Fgfr1 (a), Klb (b) and Fgf21 (c) in the eWAT of three indicated groups. (d) qRT‐PCR shows the comparison of expression levels of a battery of adipose tissue‐specific genes in the eWAT of three indicated groups. (e) The diagram shows the observed effects of short term semaglutide treatment on mice fed with HFD. In MPH, seven‐day semaglutide treatment restores the response to hFGF21 treatment. In the liver and eWAT, the treatment improves FGF21 sensitivity and restores HFD‐induced attenuation on genes that involved in maintaining lipid homeostasis and other adipose tissue‐specific genes. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Battery, Quantitative RT-PCR, Comparison, Expressing