Journal: PLOS One

Article Title: FNDC5/Irisin-dependent renoprotection of resistance training in myocardial infarction–induced Type 2 cardiorenal syndrome

doi: 10.1371/journal.pone.0342468

Figure Lengend Snippet: Myocardial infarction (MI) induces renal oxidative stress and activates the profibrotic TGF-β1/Smad2/3 signaling pathway, leading to interstitial collagen deposition and renal dysfunction. Resistance exercise upregulates renal FNDC5/Irisin expression. Irisin, potentially in concert with exercise-induced AMPK activation, attenuates oxidative stress and inhibits TGF-β1/Smad2/3 pathway activation, thereby reducing renal fibrosis and improving function. Solid lines indicate pathways supported by direct evidence from this study; dashed lines indicate proposed or associative links.

Article Snippet: Eight-week-old healthy male C57BL/6J mice (purchased from the Animal Experiment Center of Xi’an Jiaotong University) and global FNDC5/Irisin knockout mice (customized by Cyagen Biosciences Inc.) were housed under standard laboratory conditions.

Techniques: Expressing, Activation Assay

Journal: Renal Failure

Article Title: Irisin targets the HK1–glycolysis–NLRP3 pyroptosis axis to prevent chronic kidney disease-associated vascular calcification

doi: 10.1080/0886022X.2025.2610545

Figure Lengend Snippet: Fndc5 knockout aggravated aortic calcification in a CKD mouse model. (A) Experimental scheme of the CKD-associated VC mouse model. (B–E) Serum levels of creatinine (Cr), urea nitrogen (BUN), calcium (Ca), and phosphate (P) measured. *** p < 0.001, WT CKD compared with the WT group; KO CKD compared with the KO group ( n = 6). (F) Representative images of Von Kossa-stained aortic rings in different groups (positive staining: black represents calcified tissue, scale bar = 500 μm). (G) Von Kossa positive staining area of mice aortic tissue in different groups. *** p < 0.001, **** p < 0.0001 (H) Calcium content in aortic tissue from each group ( n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. indicated group; and n.s., not significant. WT: control group; KO: Fndc5 KO group; WT CKD: CKD group; KO CKD: Fndc5 KO CKD group. One-way ANOVA followed by Bonferroni’s hoc test was used for statistical analysis. Results were shown as mean ± SEM.

Article Snippet: A Fndc5 knockout cell line was constructed via the use of lentiviral vectors encoding short hairpin RNAs (shRNAs) targeting mouse Fndc5 (GeneChem Company, Shanghai, China).

Techniques: Knock-Out, Staining, Control

Journal: Renal Failure

Article Title: Irisin targets the HK1–glycolysis–NLRP3 pyroptosis axis to prevent chronic kidney disease-associated vascular calcification

doi: 10.1080/0886022X.2025.2610545

Figure Lengend Snippet: Fndc5 gene knockout promoted glycolysis and aggravated calcium deposition in β-GP-treated VSMCs. (A) Western blot analysis of Fndc5 expression in shRNA-NC VSMCs and shRNA-Fndc5 VSMCs. (B) Quantification of the results shown in (A) ( n = 3). Student’s t test was used for statistical analysis. Results were shown as mean ± SEM. * p < 0.05, vs. shRNA-NC group. (C–N) For shRNA-NC + β-GP group or shRNA-Fndc5 + β-GP group, shRNA-NC VSMCs or shRNA-Fndc5 VSMCs were treated with β-GP (10 mM) for 72 h. In the shRNA-NC + β-GP + Irisin group, shRNA-NC VSMCs were pretreated with Irisin (100 ng/ml) for 24 h before β-GP (10 mM) treatment. (C–F) Activity of glycolysis rate-limiting enzymes (HK, PFK, PK, and LDH) in culture supernatants was determined by ELISA ( n = 6). (G) Lactate levels in the culture supernatants were determined by ELISA ( n = 6). (H) Glucose uptake was determined using a 2-NBDG glucose fluorescent probe ( n = 4). Magnification: ×100; Scale bar = 50 μm; green fluorescence indicates labeled glucose molecules. (I) Quantification of the results shown in (H). (J) Glycolytic activities in VSMCs were measured using a Seahorse glycolysis assay and quantification of ECARs ( n = 3). (K–L) Glycolysis and glycolysis capacity of the results shown in (J). (M) Calcium deposition in VSMCs was detected by Alizarin red staining ( n = 3). Positive staining: red represents calcified cells; Scale bar = 100 μm. (N) Quantitative analysis of the calcium content in VSMCs using a Ca assay kit. * p < 0.05, ** p < 0.01, ** p < 0.01, *** p < 0.001, and **** p < 0.0001, vs. indicated group; and n.s., not significant. One-way ANOVA followed by Bonferroni’s hoc test was used for statistical analysis. Results were shown as mean ± SEM.

Article Snippet: A Fndc5 knockout cell line was constructed via the use of lentiviral vectors encoding short hairpin RNAs (shRNAs) targeting mouse Fndc5 (GeneChem Company, Shanghai, China).

Techniques: Gene Knockout, Western Blot, Expressing, shRNA, Activity Assay, Enzyme-linked Immunosorbent Assay, Fluorescence, Labeling, Staining

Journal: Renal Failure

Article Title: Irisin targets the HK1–glycolysis–NLRP3 pyroptosis axis to prevent chronic kidney disease-associated vascular calcification

doi: 10.1080/0886022X.2025.2610545

Figure Lengend Snippet: Irisin/FNDC5 deficiency promoted VSMC glycolysis and pyroptosis and accelerated medial calcification in CKD mice. (A) Representative images of Von Kossa-stained aortas from various groups of mice ( n = 6). Scale bar = 500 μm. (B) The protein levels of HK1, PKM2, and pyroptosis-related markers in the aortas of the different groups of mice were assessed by Western blot ( n = 6). (C) Quantification of the results shown in (B). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. indicated group; n.s., not significant. CTRL: WT mice; CKD: CKD mice; Irisin + CKD: CKD mice treated with Irisin; KO CKD: Fndc5 KO CKD mice. One-way ANOVA followed by Bonferroni’s hoc test was used for statistical analysis. Results were shown as mean ± SEM.

Article Snippet: A Fndc5 knockout cell line was constructed via the use of lentiviral vectors encoding short hairpin RNAs (shRNAs) targeting mouse Fndc5 (GeneChem Company, Shanghai, China).

Techniques: Staining, Western Blot