methyl cellulose  (Valiant Co Ltd)

Journal: Cell Death & Disease

Article Title: The triterpenoid sapogenin (2α-OH-Protopanoxadiol) ameliorates metabolic syndrome via the intestinal FXR/GLP-1 axis through gut microbiota remodelling

doi: 10.1038/s41419-020-02974-0

Figure Lengend Snippet: Six-week-old male mice were fed a high-fat diet for 8 weeks, and then treated with or without GP2-200 mg/kg/d for 5 weeks ( n = 8–10). a Body weight changes in 32 days. b Body weight gain of mice after 32 days of GP2 treatment. c Fat mass and lean mass were assessed by NMR after 4 weeks of GP2 treatment. d Organ weight of mice treated with or without GP2. e Representative pictures of H&E staining of iWAT, eWAT, BAT and the liver at the end of dietary intervention. Scale bar = 100 μm. f Levels of hepatic TG in vehicle-treated and GP2-treated mice. g – j Mice were fasted for 6 h, and the fasting blood triglyceride ( g ), cholesterol ( h ), glucose ( i ) and insulin ( j ) levels of vehicle-treated mice and GP2-treated mice were measured. k , l Oral glucose tolerance test (OGTT, 2.0 g/kg) was conducted after three weeks treatment ( k ); the area under curve of glucose level in 90 min of the OGTT ( l ). The results are shown as the mean ± s.e.m., * P < 0.05, ** P < 0.01 compared with vehicle.

Article Snippet: GP2 was dissolved in a final solution containing 0.5% methylcellulose (155496, MP Biomedicals, CA, USA), 1% dimethyl sulfoxide (DMSO; D4540-500mL, Sigma-Aldrich, Missouri, USA) and 1% Kolliphor EL (C5135-500g, Sigma-Aldrich, Missouri, USA).

Techniques: Staining

Journal: Cell Death & Disease

Article Title: The triterpenoid sapogenin (2α-OH-Protopanoxadiol) ameliorates metabolic syndrome via the intestinal FXR/GLP-1 axis through gut microbiota remodelling

doi: 10.1038/s41419-020-02974-0

Figure Lengend Snippet: a , b Immunoblot ( a ) for pro-glucagon in RIPA lysis extracts from the ilea of mice treated with or without GP2 for 2 weeks and quantification ( b ) ( n = 5). c – f Plasma concentrations of active GLP-1 ( c ) and insulin ( e ) in HFD-fed mice in response to oral of 2.0 g/kg glucose with or without GP2 treatment (200 mg/kg/d, bid) for 2 weeks ( n = 5–6); area under the curve of active GLP-1 level ( d ) and insulin level ( f ) in 15 min after glucose challenge in ( c ) and ( e ). Results are shown as the mean ± s.e.m., * P < 0.05, ** P < 0.01 compared with vehicle.

Article Snippet: GP2 was dissolved in a final solution containing 0.5% methylcellulose (155496, MP Biomedicals, CA, USA), 1% dimethyl sulfoxide (DMSO; D4540-500mL, Sigma-Aldrich, Missouri, USA) and 1% Kolliphor EL (C5135-500g, Sigma-Aldrich, Missouri, USA).

Techniques: Western Blot, Lysis

Journal: Cell Death & Disease

Article Title: The triterpenoid sapogenin (2α-OH-Protopanoxadiol) ameliorates metabolic syndrome via the intestinal FXR/GLP-1 axis through gut microbiota remodelling

doi: 10.1038/s41419-020-02974-0

Figure Lengend Snippet: a , b mRNA expression of the target genes in the ileum ( a ) and liver ( a , b ) ( n = 7–8). c Gene expression of the target genes in the ileum ( n = 7–8). d Individual taurine-conjugated bile acid levels were detected in the feces of HFD-fed mice ( n = 3). e BSH enzymatic activity in the feces of HFD-fed mice treated with or without GP2 (200 mg/kg). The ratio of CDCA-d5 to TCDCA-d5 was used to represent the enzymatic activity of BSH ( n = 4). f BSH enzymatic activity in the feces of HFD-fed mice treated with different concentrations of GP2 in vitro. The ratio of CDCA-d5 to TCDCA-d5 was used to represent the enzymatic activity of BSH ( n = 3). g GLP-1 concentration in the supernatants of STC-1 cells were measured after incubation with DMSO or TβMCA for 24 h, and the TGR5 agonist INT-777 (10 μM) was used as a positive control ( n = 4). h Gene expression of the target genes in STC-1 cells treated with 300 μM TβMCA for 12 h ( n = 4). i Quantitation of the extracellular acidification rate (ECAR) by the seahorse assay. STC-1 cells were incubated with DMSO or TβMCA for 24 h and then successively injected with glucose (10 mM), oligomycin (1 μM), 2-deoxyglucose (100 mM) and rotenone (1 μM)/antimycin A (1 μM) for the seahorse flux assay at the indicated time. j – l Eight-week-old male WT or intestinal specific FXR KO (FXR −/− ) mice were fed a high-fat diet for 5 weeks, and then treated with or without GP2-200 mg/kg/d for 2 weeks ( n = 5–6). Intestinal Fxr mRNA levels ( j ) and pro-glucagon protein levels ( k , l ) from WT mice and FXR −/− mice after GP2 treatment were detected. The results are shown as the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001 compared with vehicle or DMSO.

Article Snippet: GP2 was dissolved in a final solution containing 0.5% methylcellulose (155496, MP Biomedicals, CA, USA), 1% dimethyl sulfoxide (DMSO; D4540-500mL, Sigma-Aldrich, Missouri, USA) and 1% Kolliphor EL (C5135-500g, Sigma-Aldrich, Missouri, USA).

Techniques: Expressing, Activity Assay, In Vitro, Concentration Assay, Incubation, Positive Control, Quantitation Assay, Injection, Flux Assay

Journal: Cell Death & Disease

Article Title: The triterpenoid sapogenin (2α-OH-Protopanoxadiol) ameliorates metabolic syndrome via the intestinal FXR/GLP-1 axis through gut microbiota remodelling

doi: 10.1038/s41419-020-02974-0

Figure Lengend Snippet: Six-week-old male mice were fed a high-fat diet for 8 weeks, and then treated with or without 200 mg/kg/d GP2 for 5 weeks ( n = 4). a , b Sobs index analysis of the gut microbiome of HFD-fed mice after oral administration of GP2 for 5 weeks. c Principal coordinate analysis of gut microbiota composition based on unweighted UniFrac. d Pie charts of the average relative abundance of the gut microbiota in the feces at the phylum level. e Heatmap depicting the relative abundance of the microbiota in the feces at the genus level as in d . The results are shown as the mean ± s.e.m., *** P < 0.001 compared with vehicle.

Article Snippet: GP2 was dissolved in a final solution containing 0.5% methylcellulose (155496, MP Biomedicals, CA, USA), 1% dimethyl sulfoxide (DMSO; D4540-500mL, Sigma-Aldrich, Missouri, USA) and 1% Kolliphor EL (C5135-500g, Sigma-Aldrich, Missouri, USA).

Techniques:

Journal: Cell Death & Disease

Article Title: The triterpenoid sapogenin (2α-OH-Protopanoxadiol) ameliorates metabolic syndrome via the intestinal FXR/GLP-1 axis through gut microbiota remodelling

doi: 10.1038/s41419-020-02974-0

Figure Lengend Snippet: a Schematic diagram of the experiment ( n = 9–11). In brief, 6-week-old male mice were fed a high-fat diet for 10 weeks, and then the mice were randomly divided into four groups according to body weight and fasting blood glucose levels. Two groups of mice were treated with an antibiotic mix (ABX) for 1 week and then treated with or without GP2 combined with ABX for 3 weeks; the other two groups of mice were fed a high-fat diet for 1 week, and then treated with or without GP2 for 3 weeks. b Fasting blood glucose concentrations were measured after treatment with or without GP2 (200 mg/kg/d), and treatment with or without ABX for 3 weeks. The animals were fasted for 6 h before blood glucose determination. c The oral glucose tolerance test (OGTT) results of vehicle-, GP2-, vehicle+ ABX-and GP2+ABX-treated mice after glucose (2.0 g/kg) administration by gavage were plotted. d Area under the curve of glucose level in 90 min of OGTT in ( c ), e , f Quantitative mRNA expression of pro-glucagon in the ileum ( e ) and colon ( f ) after GP2 treatment combined with or without ABX treatment. The results are shown as the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001 compared with vehicle.

Article Snippet: GP2 was dissolved in a final solution containing 0.5% methylcellulose (155496, MP Biomedicals, CA, USA), 1% dimethyl sulfoxide (DMSO; D4540-500mL, Sigma-Aldrich, Missouri, USA) and 1% Kolliphor EL (C5135-500g, Sigma-Aldrich, Missouri, USA).

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: The triterpenoid sapogenin (2α-OH-Protopanoxadiol) ameliorates metabolic syndrome via the intestinal FXR/GLP-1 axis through gut microbiota remodelling

doi: 10.1038/s41419-020-02974-0

Figure Lengend Snippet: a Schematic diagram of the pharmacodynamic experiment. In brief, 6-week-old male mice were fed a high-fat diet for 20 weeks, and then the gut microbiota was depleted by antibiotics in drinking water for 3 days. Twelve hours after last dose of antibiotics given, the mice were oral administered the feces (resuspended in PBS) isolated from vehicle (Re-Veh) or GP2-treated mice (Re-GP2) for three times (on day 0, day 3, day 6). Then the mice were fed a high-fat diet for another 3 weeks. b The mice were fasted for 6 h and an oral glucose tolerance test (OGTT) was conducted after glucose (2.0 g/kg) after administration by gavage ( n = 8–10). c Area under the curve of glucose level in 90 min of OGTT in b ( n = 8–10). d The levels of the individual taurine-conjugated bile acids TαMCA, TβMCA, and TUDCA were measured in the feces ( n = 5–6). e Plasma concentration of active GLP-1 in Re-Veh and Re-GP2 mice in response to oral administration of 2.0 g/kg glucose ( n = 5–6). f Area under the curve of active GLP-1 level in 15 min after glucose challenge in e . g , h Immunoblot ( g ) of pro-glucagon in RIPA lysis extracts from the ilea of Re-Veh and Re-GP2 mice and the quantification ( h ) ( n = 5). The results are shown as the mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001 compared with Re-Veh.

Article Snippet: GP2 was dissolved in a final solution containing 0.5% methylcellulose (155496, MP Biomedicals, CA, USA), 1% dimethyl sulfoxide (DMSO; D4540-500mL, Sigma-Aldrich, Missouri, USA) and 1% Kolliphor EL (C5135-500g, Sigma-Aldrich, Missouri, USA).

Techniques: Isolation, Concentration Assay, Western Blot, Lysis