Journal: Advanced Science

Article Title: Trigonelline Improves Metabolism and Cardiac Function of HFpEF Mice Via Gut Microbiome Alterations‐Mediated AMPK Activation

doi: 10.1002/advs.202513956

Figure Lengend Snippet: AMPK activation was required for the cardioprotective effects of trigonelline in HFpEF mice. A) Representative immunoblot images of total and phosphorylated AMPK in the heart and liver tissues from HFpEF mice receiving trigonelline. B,C) Quantification of cardiac p‐AMPK/AMPK ( n = 6 mice per group). D) Representative immunoblot images of total and phosphorylated AMPK in the heart and liver tissues from HFpEF mice receiving trigonelline with or without AMPK inhibitor. E,F) Quantification of hepatic p‐AMPK/AMPK ratio ( n = 6 mice per group). G,H) SBP and DBP of different experimental groups ( n = 5 mice per group). I) Representative echocardiography‐derived M‐mode tracings (top), pulsed‐wave Doppler (middle), and tissue Doppler (bottom) tracings of mice in the indicated group. J) Percent left ventricular ejection fraction (LVEF%). K) The ratio between mitral E wave and E’ wave (E/E’). L) Running distance during the exercise exhaustion test of mice. M) The ratio between wet and dry lung weight (LW). N) Ratio between heart weight and tibia length (HW/TL) (for LVEF%, E/E’ ratio, running distance, LW wet/LW dry ratio, and HW/TL ratio, n = 6 mice per group). O) Representative images of WGA in transversal sections of the left ventricle of mice of different experimental groups. Scale bar: 50 µm for WGA. P) WGA quantification of cardiomyocyte cross‐sectional area ( n = 5 mice per group). Data are presented as mean ± SEM and analyzed using one‐way ANOVA followed by Tukey's multiple comparisons test. ns, no significant; * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: To test the role of AMPK in the therapeutic effects of trigonelline in vivo, HFpEF mice were pre‐treated with AMPK inhibitor Compound C (CC) (MedChemExpress, HY‐13418, 5 mg kg −1 day −1 ) or vehicle for 1 week.

Techniques: Activation Assay, Western Blot, Derivative Assay

Journal: Advanced Science

Article Title: Trigonelline Improves Metabolism and Cardiac Function of HFpEF Mice Via Gut Microbiome Alterations‐Mediated AMPK Activation

doi: 10.1002/advs.202513956

Figure Lengend Snippet: AMPK inhibition abolished the beneficial effects of trigonelline on metabolic disorders in HFpEF mice. A) Food intake of mice per day of different experimental groups per day ( n = 5). B) Body weight was monitored weekly in each experimental group. C) Representative images of MRI of mice from different experimental groups. Red indicates higher fat content, green represents moderate fat content, and blue corresponds to low fat content. Scale bar: 1 cm. Fat mass D) and lean mass E) ratios of mice in the indicated groups ( n = 5). F) Glucose tolerance tests in the indicated groups ( n = 5). G) Insulin sensitivity tests in the indicated groups ( n = 5). H) Total cholesterol of serum in each group. I) Low‐density lipoprotein cholesterol (LDL‐C) of the serum in each group. J) Representative images of hematoxylin and eosin staining and oil red staining of liver from mice in each treated group, Scale bar = 100 µm. K) Serum ALT in each group. L) Serum AST in each group. M) Serum Cre in each group. N) Serum BUN in each group (for total cholesterol, LDL‐C, Serum ALT, AST, Cre, and BUN, n = 5 mice per group). Data are presented as mean ± SEM and analyzed using one‐way ANOVA followed by Tukey's multiple comparisons test. ns, no significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: To test the role of AMPK in the therapeutic effects of trigonelline in vivo, HFpEF mice were pre‐treated with AMPK inhibitor Compound C (CC) (MedChemExpress, HY‐13418, 5 mg kg −1 day −1 ) or vehicle for 1 week.

Techniques: Inhibition, Staining

Journal: Advanced Science

Article Title: Trigonelline Improves Metabolism and Cardiac Function of HFpEF Mice Via Gut Microbiome Alterations‐Mediated AMPK Activation

doi: 10.1002/advs.202513956

Figure Lengend Snippet: Trigonelline activates AMPK in a gut microbiota‐dependent manner. A–C) Diversity of the gut microbiota in each group, as indicated by the observed species, Shannon, and Chao1 indices. D) PCA score plot analysis based on the relative abundance of OTUs. E) Scheme for the experimental strategy in HFpEF mice treated with trigonelline and antibiotics. F) Representative immunoblot images of total and phosphorylated AMPK in the heart and liver tissues of mice in each group. Quantification of cardiac pAMPK/AMPK ratio in the heart G) and liver H) tissues of mice in each group. ( n = 6 mice per group). I) Representative immunoblot images of total and phosphorylated AMPK in HL‐1 and HepG2 cell lines in each group. Quantification of cardiac pAMPK/AMPK ratio in the HL‐1 J) and HepG2 cell lines K) in each group. ( n = 4 mice per group). Data are presented as mean ± SEM and analyzed using one‐way ANOVA followed by Tukey's multiple comparisons test. ns, no significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. Abx, antibiotic cocktail.

Article Snippet: To test the role of AMPK in the therapeutic effects of trigonelline in vivo, HFpEF mice were pre‐treated with AMPK inhibitor Compound C (CC) (MedChemExpress, HY‐13418, 5 mg kg −1 day −1 ) or vehicle for 1 week.

Techniques: Western Blot