Journal: Scientific Reports

Article Title: Voluntary exercise in mice fed an obesogenic diet alters the hepatic immune phenotype and improves metabolic parameters – an animal model of life style intervention in NAFLD

doi: 10.1038/s41598-018-38321-9

Figure Lengend Snippet: The effects of diet and running wheel access on body weight, caloric intake and physical activity. C57BL/6J mice received a high-fat, high-carbohydrate diet (HFD) or corresponding control diet (CD) for 12 weeks. After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD. Data represent means ± SEM of ( A–H ) n = 7 VWR/CD, ( A–H ) n = 8 VWR/HFD, ( A , B , D and E ) n = 11 SED/CD and ( A , B , D and E ) n = 20 SED/HFD mice. *p < 0.05, **p < 0.01, ***p < 0.001 for CD vs. HFD and $ p < 0.05, $$$ p < 0.001 for SED vs. VWR using Mann-Whitney U test ( A,F–H ) and two-tailed Student’s t -test ( B – E ).

Article Snippet: After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD.

Techniques: Activity Assay, MANN-WHITNEY, Two Tailed Test

Journal: Scientific Reports

Article Title: Voluntary exercise in mice fed an obesogenic diet alters the hepatic immune phenotype and improves metabolic parameters – an animal model of life style intervention in NAFLD

doi: 10.1038/s41598-018-38321-9

Figure Lengend Snippet: Effects of VWR on liver enzymes and metabolic parameters. Fasting levels of glucose, triglycerides, total cholesterol and ALT of CD- and HFD-fed mice were assessed ( A ) pre-exercise and ( B ) post-exercise in comparison with diet-matched SED groups. ( C ) Fasting insulin levels and ( D ) HOMA-IR (fasting insulin concentration (pmol/l) x fasting glucose concentration (mmol/l)/22.5) in the VWR and SED groups after 12 weeks of feeding. Data are means ± SEM of ( A – D ) n = 7 VWR/CD, ( A – D ) n = 8 VWR/HFD, ( B – D ) n = 11 SED/CD and ( B – D ) n = 20 SED/HFD mice. ***p < 0.001 for CD vs. HFD and $$ p < 0.01 for SED vs. VWR using two-tailed Student’s t -test ( A and B (fasting glucose, total cholesterol)) or Mann-Whitney U test ( B (ALT)- D ).

Article Snippet: After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD.

Techniques: Concentration Assay, Two Tailed Test, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Voluntary exercise in mice fed an obesogenic diet alters the hepatic immune phenotype and improves metabolic parameters – an animal model of life style intervention in NAFLD

doi: 10.1038/s41598-018-38321-9

Figure Lengend Snippet: Effects of VWR on hepatic lipid metabolism and steatosis in HFD-fed mice. ( A ) Absolute liver weight, ( B ) liver triglyceride content, ( C ) representative liver histology by H&E staining (scale bar: 200 µm) and NAFLD activity score (NAS), and ( D ) relative mRNA expression of key regulators of hepatic lipid metabolism determined by qRT-PCR in VWR and SED mice after 12 weeks of dietary feeding. Data represent means ± SEM of ( A ) n = 7 VWR/CD, n = 8 VWR/HFD, n = 11 SED/CD, n = 20 SED/HFD, ( B ) n = 5 mice/group, ( C ) n = 7 VWR/CD, n = 8 VWR/HFD, n = 4 SED/CD, n = 8 SED/HFD, ( D ) n = 7 VWR/CD, n = 8 VWR/HFD, n = 5 SED/CD and n = 5 SED/HFD mice. *p < 0.05, **p < 0.01, ***p < 0.001 for CD vs. HFD and $ p < 0.05, $$ p < 0.01 for SED vs. VWR using Mann-Whitney U test ( A , C ) or two-tailed Student’s t -test ( D ).

Article Snippet: After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD.

Techniques: Staining, Activity Assay, Expressing, Quantitative RT-PCR, MANN-WHITNEY, Two Tailed Test

Journal: Scientific Reports

Article Title: Voluntary exercise in mice fed an obesogenic diet alters the hepatic immune phenotype and improves metabolic parameters – an animal model of life style intervention in NAFLD

doi: 10.1038/s41598-018-38321-9

Figure Lengend Snippet: Effects of VWR on AMPK-α activation and glucose metabolism in the liver. ( A ) Western blot and densitometric analyses of phospho-AMPK-α-Thr172 and total AMPK-α protein in liver lysates from VWR vs. SED mice fed the diets for 12 weeks and ( B ) from VWR mice on CD or HFD with increasing individual physical activity levels (increasing from left to right). ( C ) qRT-PCR analyses of gluconeogenetic enzymes and ( D ) Western blot with densitometric analysis of phosphorylated (Ser473) and total Akt protein in the liver of the different experimental groups after 12 weeks of feeding. In A, B and D representative immunoblots with densitometric analysis are shown. Uncropped images of original blots are shown in Suppl. Fig. . Data in C represent means ± SEM of n = 7 VWR/CD, n = 8 VWR/HFD, n = 5 SED/CD and n = 5 SED/HFD mice. *p < 0.05 for CD vs. HFD and $ p < 0.05, $$ p < 0.01 for SED vs. VWR using two-tailed Student’s t -test ( A – D ).

Article Snippet: After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD.

Techniques: Activation Assay, Western Blot, Activity Assay, Quantitative RT-PCR, Two Tailed Test

Journal: Scientific Reports

Article Title: Voluntary exercise in mice fed an obesogenic diet alters the hepatic immune phenotype and improves metabolic parameters – an animal model of life style intervention in NAFLD

doi: 10.1038/s41598-018-38321-9

Figure Lengend Snippet: Impact of VWR on HFD-induced hepatic inflammation and macrophage infiltration. ( A ) Relative mRNA expression levels in whole liver of pro-inflammatory and pro-fibrotic cytokines, and ( B ) the macrophage-marker F4/80, ( C ) relative numbers of living, intrahepatic CD45 + F4/80 + cells quantified by flow cytometry, and ( D ) CCL2 serum levels in VWR and SED mice after 12 weeks of exposure to CD or HFD. Data in A and B are means ± SEM of ( A , B ) n = 7 VWR/CD, n = 8 VWR/HFD, n = 5 SED/CD, n = 5 SED/HFD mice, ( C , D ) n = 5 VWR/CD, n = 5 VWR/HFD, n = 5 SED/CD and n = 15 SED/HFD mice. *p < 0.05 for CD vs. HFD and $ p < 0.05 for SED vs. VWR using Mann-Whitney U test ( A , C ) or two-tailed Student’s t -test ( B,D ).

Article Snippet: After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD.

Techniques: Expressing, Marker, Flow Cytometry, MANN-WHITNEY, Two Tailed Test

Journal: Scientific Reports

Article Title: Voluntary exercise in mice fed an obesogenic diet alters the hepatic immune phenotype and improves metabolic parameters – an animal model of life style intervention in NAFLD

doi: 10.1038/s41598-018-38321-9

Figure Lengend Snippet: Effects of VWR on NF-κB activation in the liver of HFD-fed mice. ( A ) Phospho-NF-κB p65 (Ser536) and total NF-κB p65 protein expression were determined in liver lysates from VWR mice compared with SED mice and ( B ) from VWR mice with increasing individual physical activity levels (increasing from left to right) fed the diets for 12 weeks by western blot, and ( C ) activated p65 was quantified by a specific DNA-binding ELISA. Data are shown as fold change relative to the mean of SED/CD mice, which was considered 1. In ( A,B ) representative immunoblots with densitometric analysis are shown. Uncropped images of original blots are shown in Suppl. Fig. . Data in C are means ± SEM of n = 5 mice/group. *p < 0.05, **p < 0.01 for CD vs. HFD using two-tailed Student’s t -test ( A – C ).

Article Snippet: After 8 weeks of dietary feeding, mice were randomly assigned to either a group given access to voluntary running wheels (VWR) for 4 weeks or a sedentary control group (SED). ( A ) Body weight curve of the VWR/CD, VWR/HFD, SED/CD, and SED/HFD group was monitored and ( B ) % change in body weight in the VWR groups during the 4 week-exercise period was compared with diet-matched SED groups. ( C ) Caloric intake of the VWR/CD and VWR/HFD group during the pre-exercise, during-exercise and ( D ) whole experimental period compared with SED controls and ( E ) ratio of average caloric intake to final body weight for all experimental groups. ( F ) Cumulative running distance, ( G ) average daily running distance, and ( H ) ratio of total running distance to final body weight for mice fed the CD or HFD.

Techniques: Activation Assay, Expressing, Activity Assay, Western Blot, Binding Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Applied Physiology

Article Title: Voluntary wheel running prevents salt-induced endothelial dysfunction: role of oxidative stress

doi: 10.1152/japplphysiol.00421.2018

Figure Lengend Snippet: Endothelium-dependent relaxation (EDR) in sedentary and voluntary wheel running (VWR) rats fed either a normal-salt (NS) or a high-salt (HS) diet: dose responses of femoral rings to acetylcholine (ACh) under control (solid lines) and NG-nitro-l-arginine methyl ester (l-NAME)-treated (dashed lines) conditions (A), maximal relaxation (Emax) to ACh (B), and area under the curve (AUC) to ACh (C). Values are means ± SE; n: NS = 12, NS-VWR = 13, HS = 14, HS-VWR = 12. Data analysis was performed with a 1-way ANOVA with a Tukey’s post hoc test. *vs. HS, α vs. NS, †vs. NS-VWR, ‡vs. HS-VWR, §vs. respective control group (P < 0.05).

Article Snippet: Values are means ± SE; n : NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12.

Techniques:

Journal: Journal of Applied Physiology

Article Title: Voluntary wheel running prevents salt-induced endothelial dysfunction: role of oxidative stress

doi: 10.1152/japplphysiol.00421.2018

Figure Lengend Snippet: Endothelium-independent relaxation (EIR) in sedentary and voluntary wheel running (VWR) rats fed either a normal-salt (NS) or a high-salt (HS) diet. Values are means ± SE; n: NS = 12, NS-VWR = 13, HS = 14, HS-VWR = 12. Data analysis was performed with a 1-way ANOVA with a Tukey’s post hoc test. SNP, sodium nitroprusside.

Article Snippet: Values are means ± SE; n : NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12.

Techniques:

Journal: Journal of Applied Physiology

Article Title: Voluntary wheel running prevents salt-induced endothelial dysfunction: role of oxidative stress

doi: 10.1152/japplphysiol.00421.2018

Figure Lengend Snippet: A–C: total endothelial nitric oxide synthase (eNOS) protein expression in the femoral artery normalized to β-actin (A), phosphorylated eNOS (eNOS-P, ser1177) relative to total eNOS (B), and representative Western blots (C). D–F: total eNOS protein expression in the aorta normalized to β-actin (D), eNOS-P (ser1177) relative to total eNOS (E), and representative Western blots (F). Values are means ± SE; n = 9 or 10 per group. Data analysis was performed with a 1-way ANOVA with a Tukey’s post hoc test. HS, high salt; NS, normal salt; Sed, sedentary; VWR, voluntary wheel running. *vs. HS, †vs. NS-VWR (P < 0.05).

Article Snippet: Values are means ± SE; n : NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12.

Techniques: Expressing, Western Blot

Journal: Journal of Applied Physiology

Article Title: Voluntary wheel running prevents salt-induced endothelial dysfunction: role of oxidative stress

doi: 10.1152/japplphysiol.00421.2018

Figure Lengend Snippet: Femoral nitrotyrosine expression normalized to β-actin (A), total aortic NOX4 protein expression normalized to β-actin (B), aortic Gp91phox normalized to β-actin (C), and representative Western blots (D). Values are means ± SE; n = 8 or 9 per group. Data analysis was performed with a 1-way ANOVA with a Tukey’s post hoc test. HS, high salt; NS, normal salt; VWR, voluntary wheel running. α vs. NS, †vs. NS-VWR, ‡ vs. HS-VWR (P < 0.05).

Article Snippet: Values are means ± SE; n : NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12.

Techniques: Expressing, Western Blot

Journal: Journal of Applied Physiology

Article Title: Voluntary wheel running prevents salt-induced endothelial dysfunction: role of oxidative stress

doi: 10.1152/japplphysiol.00421.2018

Figure Lengend Snippet: Superoxide dismutase-1 (SOD-1) protein expression in the femoral artery relative to β-actin (A), total SOD-2 protein expression in the aorta normalized to β-actin (B), representative Western blots of SOD-1 and SOD-2 (C), catalase protein expression in the aorta normalized to β-actin (D), GPx-1/2 expression in the aorta normalized to β-actin (E), and representative Western blots of catalase and GPx-1/2 (F). Values are means ± SE; n = 7 or 8 per group. Data analysis was performed with a 1-way ANOVA with a Tukey’s post hoc test. HS, high salt; NS, normal salt; VWR, voluntary wheel running. *vs. HS, α vs. NS (P < 0.05).

Article Snippet: Values are means ± SE; n : NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12.

Techniques: Expressing, Western Blot

Journal: Journal of Applied Physiology

Article Title: Voluntary wheel running prevents salt-induced endothelial dysfunction: role of oxidative stress

doi: 10.1152/japplphysiol.00421.2018

Figure Lengend Snippet: A: endothelium-dependent relaxation (EDR) in the femoral artery in normal-salt (NS) rats in response to acetylcholine (ACh) with and without addition of the superoxide dismutase mimetic TEMPOL. B: EDR in the femoral artery in high-salt (HS) rats in response to ACh with and without the addition of TEMPOL. C: EDR in the femoral artery in NS rats in response to ACh with and without addition of the NADPH oxidase inhibitor apocynin (APO). D: EDR in femoral artery in HS rats in response to ACh with and without the addition of the NADPH oxidase inhibitor APO. E: EDR in the femoral artery in NS rats in response to ACh with and without addition of TEMPOL + APO (T/A). F: EDR in femoral artery in HS rats in response to ACh with and without the addition of T/A. G: maximal relaxation with and without the addition of TEMPOL, APO, or T/A. H: area under the curve (AUC) with and without the addition of TEMPOL, APO, or T/A. Values are means ± SE; n: NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12. Data analysis was performed with a (4 × 3) 2-way repeated-measures ANOVA.*vs. HS, §vs. respective control group (P < 0.05).

Article Snippet: Values are means ± SE; n : NS = 12, NS-voluntary wheel running (VWR) = 13, HS = 14, HS-VWR = 12.

Techniques: