ptr6330-sup-0001-FigureS1.docxWord 2007 document , 4.3 MB

Figure S1 │ Guarana did not affect hepatic fat accumulation (hepatosteatosis) measured by histology. (A) Representative liver sections stained with H&E. (B) Graphical representation of Chi‐square test from steatosis score. Significant differences (n = 4) are indicated by * p < 0.05. Figure S2 │ Neither WD nor guarana had any effect on cholesterol or the inflammatory profile. (A) Fasting HDL (mg/dL). (B) Fasting LDL (mg/dL). (C) Serum IL‐6 (pg/mL). (D) Serum TNF‐α/IL‐10 ration. Data are expressed as means ± SEM. Significant differences (n = 8) are indicated by * p < 0.05 (ANOVA followed by Tukey between Control, WD, and WD + Gua groups). Figure S3│ Guarana supplementation used in this study was not toxic. (A‐D) The guarana dose supplemented to WD (0.5%) did not lead to liver damage and renal function alteration in the animals. (A) Liver protein damage expressed as nmol carbonyl groups/mg protein. (B) Liver lipip peroxidation level expressed as nmol thiobarbituric acid reactive substances (TBARS)/mg protein. (C) Serum aspartate aminotransferase level (U/L). (D) Serum alanine aminotransferase level (U/L). (E‐F) Nephrotoxicity evaluation of guarana dose. (E) Serum creatinine level (mg/dL). (F) Serum uric acid level (mg/dL). (G‐H) Protein damage and Lipip peroxidation level on cortex, hippocampus, and hypothalamus. Data are expressed as means ± SEM. Significant differences (n = 8) are indicated by * p < 0.05 and ** p < 0.01 (ANOVA followed by Tukey). (I‐P) Guarana supplementation (0.5%) did not induce behavior changes or memory impairment in Wistar rats. (I‐L)Emotional behavioral parameters in open‐field test: total distance as a measure of horizontal exploratory activity (I), rearing time as a measure of vertical exploratory activity (J), center time as a measure of anxiety‐like behavior (K), and freezing time as a measure of behavioral inhibition (L). (M‐P) Behavioral parameters in learning and memory tasks: (M) % of spontaneous alternation and (N) total entries in arms in the Y‐maze test as a measure of spatial working memory, and (O‐P) time of novel object exploration 4 h (O) and 24 h (P) after object familiarization in the novel object recognition task as a measure of short and long‐term recognize memory, respectively. Data are expressed as means ± SEM. Significant differences (n = 12) are indicated by * p < 0.05 (ANOVA followed by Tukey). Figure S4│Guarana induces minor changes in the gut microbiota. (A) Calculations of within‐community richness (α‐diversity) by Chao1 estimate. (B) Calculation of ration between Firmicutes and Bacteroidetes phyla. (A‐B) Data are expressed as means ± SEM. Significant differences (n = 4) are indicated by * p < 0.05 (ANOVA followed by Tukey). (C‐E) Relative abundances of statistically different genera: (C) Butyricicoccus, (D) Holdemania, and (E) Streptococcus. (F) Venn diagram showing presence of unique genera of each group. (G‐H) Relative abundances of statistically different species: (C) Bacteroides uniformis and (H) Bacteroides eggerthii. (C‐E and G‐H) The box‐plot shows the median of the data as a line, the mean of the data as a star, and data points outside of the whiskers as crosses. Significant differences (n = 4) are indicated by * p < 0.05 and ** p < 0.01 (ANOVA followed by Tukey–Kramer post hoc).