Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: (A) Celluar study design. (B) After SIRT1 overexpression, western blotting was performed on H9c2 cells to determine PGC1α, PPARα, PDK4, CPT1, GPX4, and SLC7A11 levels, and grayscale values were analyzed, with β-actin serving as a protein loading control. Intracellular levels of FFA (C) and A-CoA (D) after SIRT1 overexpression in H9c2 cells. (E) Intracellular ATP levels in H9c2 cells after SIRT1 overexpression. (F) Intracellular GSH/GSSG levels in H9c2 cells after SIRT1 overexpression. (G) Representative images of TMRE staining and quantitative analysis of fluorescence (I) in H9c2 cells after SIRT1 overexpression. Scale bar = 100 μm. (H) Representative images of DCFH-DA staining and quantitative analysis of fluorescence (J) in H9c2 cells after SIRT1 overexpression. Scale bar = 100 μm. (K) CCK-8 assays were employed to assess the viability of H9c2 cells after SIRT1 overexpression. Statistical significance was indicated as *P < 0.05, **P < 0.01 vs. Control, #P < 0.05, ##P < 0.01 vs. Hypoxia.

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques: Over Expression, Western Blot, Control, Staining, Fluorescence, CCK-8 Assay

Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: (A) Celluar study design. (B) After overexpression of SIRT1, PPARα was inhibited by GW6471 in H9c2 cells. Western blotting was conducted to measure the levels of PGC1α, PPARα, PDK4, CPT1, GPX4, and SLC7A11 in H9c2 cardiomyocytes, and grayscale values were analyzed. β-actin was used as a protein loading control. Intracellular levels of FFA (C), A-CoA (D), ATP (E), and GSH/GSSG (F) in H9c2 cells after SIRT1 overexpression and PPARα inhibition. (G) Representative images of TMRE staining and quantitative analysis of fluorescence (I) in H9c2 cells after SIRT1 overexpression and PPARα inhibition. Scale bar = 100 μm. (H) Representative images of DCFH-DA staining and quantitative analysis of fluorescence (J) in H9c2 cells after SIRT1 overexpression and PPARα inhibition. Scale bar = 100 μm. (K) CCK-8 assays were employed to assess the viability of H9c2 cells after SIRT1 overexpression and PPARα inhibition. Statistical significance was indicated as *P < 0.05, **P < 0.01 vs. Control, #P < 0.05, ##P < 0.01 vs. Hypoxia, &P < 0.05, &&P < 0.01 vs. Hypoxia+SIRT1 OE .

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques: Over Expression, Western Blot, Control, Inhibition, Staining, Fluorescence, CCK-8 Assay

Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: (A) Animal study design. (B) Echocardiography in rats. (C-E) RSV decreased LVPWd and LVPWs levels and increased LVEDV, LVESV, SV, and CO levels of the heart in the HH test. (F) RSV decreased the cardiac coefficient compared with the HH group. (G) Transmission electron microscopy was utilized to visualize the left ventricular myocardium ultrastructure in rats (8,000×). Scale bar = 1 μm. (H, I) A detailed analysis was conducted to determine the number of mitochondria per unit area and the average mitochondrial size. Myocardial FFA (J), A-CoA (K), and ATP (L) levels in rats after treatment with RSV. (M) DHE staining of rat myocardial tissue (40×) after RSV treatment and quantitative analysis of fluorescence (O). (N) Western blotting was performed on rat myocardial tissue extracts to determine SIRT1, PGC1α, PPARα, PDK4, CPT1, GPX4, and SLC7A11 levels, and grayscale values were analyzed (Q), with β-actin serving as a protein loading control. (P) Myocardial GSH/GSSG levels in rats after treatment with RSV. Statistical significance was indicated as *P < 0.05, **P < 0.01 vs. Control, #P < 0.05, ##P < 0.01 vs. HH.

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques: Transmission Assay, Electron Microscopy, Staining, Fluorescence, Western Blot, Control

Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: Under conditions of HH exposure, the downregulation of SIRT1 triggers FAO and ferroptosis through the PPARα-PDK4/CPT1-GPX4/SLC7A11 pathway. This results in mitochondrial dysfunction, ultimately contributing to the development of cardiac impairment, as depicted in the pathway map.

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Cordycepin Ameliorates Constant Light-Induced Thermogenic Dysfunction in Brown Adipose Tissue by Activating SIRT1-Mediated Mitochondrial Homeostasis

doi: 10.3390/ijms27104351

Figure Lengend Snippet: Cordycepin enhances the SIRT1-PGC-1 α signaling axis via a dual mechanism involving direct binding and indirect modulation of NAD + . ( A , C ) Representative Western blots of SIRT1, acetylated PGC-1 α (Ac-PGC-1 α ), and total PGC-1 α in BAT lysates; ( B , D – E ) densitometric quantification of SIRT1,Ac-PGC-1 α and total PGC-1 α ; ( F ) representative Western blot and densitometric quantification of P-AMPK/AMPK protein levels; ( G , H ) in silico molecular docking simulations illustrating the predicted binding pose of Cpn within the catalytic pocket of human SIRT1 with key interacting residues highlighted; ( I , J ) Surface Plasmon Resonance (SPR) analysis showing the direct physical interaction between Cpn and recombinant human SIRT1 protein with representative sensorgram ( I ) and steady-state affinity analysis ( J ) shown. The equilibrium dissociation constant (KD) was calculated; ( K – M ) intracellular NAD + /NADH ratio ( M ) and SIRT1 deacetylase activity ( K , L ), measured in primary brown adipocytes treated with Cpn (Cpn, 10 μM) and/or the SIRT1-specific inhibitor EX-527 (10 μM) for 48 h. Tubulin was used as a loading control for Western blots. Data are presented as mean ± SD with individual data points shown ( n = 3 mice per group for Western blot; n = 3 independent experiments for in vitro studies). Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Recombinant human SIRT1 (MedChemExpress, HY- P71596 , Monmouth Junction, NJ, USA) was immobilized on a CM5 sensor chip via amine coupling using the standard EDC/NHS chemistry.

Techniques: Binding Assay, Western Blot, In Silico, SPR Assay, Recombinant, Histone Deacetylase Assay, Activity Assay, Control, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Cordycepin Ameliorates Constant Light-Induced Thermogenic Dysfunction in Brown Adipose Tissue by Activating SIRT1-Mediated Mitochondrial Homeostasis

doi: 10.3390/ijms27104351

Figure Lengend Snippet: The inhibition of SIRT1 abrogates the beneficial effects of cordycepin on mitochondrial function and metabolic reprogramming in brown adipocytes. All experiments were performed on cultured primary brown adipocytes. Cells were pre-treated with cordycepin (Cpn, 10 μM) and/or the SIRT1-specific inhibitor EX-527 (10 μM) for 48 h. ( A ) Cell viability determined by CCK8 assay after treatment with increasing concentrations of Cpn; ( B – E ) dose–response analysis of Cpn on key target genes to determine its optimal working concentration; ( F – J ) quantification of cellular oxidative stress markers, including mitochondrial ROS (mtROS) ( H – J ) Scale bar: 200 μm, malondialdehyde (MDA) ( F ), and superoxide dismutase (SOD) activity ( G ). Data are presented as mean ± SD with individual data points shown ( n = 3 independent experiments for in vitro studies). Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Recombinant human SIRT1 (MedChemExpress, HY- P71596 , Monmouth Junction, NJ, USA) was immobilized on a CM5 sensor chip via amine coupling using the standard EDC/NHS chemistry.

Techniques: Inhibition, Cell Culture, CCK-8 Assay, Concentration Assay, Activity Assay, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Cordycepin Ameliorates Constant Light-Induced Thermogenic Dysfunction in Brown Adipose Tissue by Activating SIRT1-Mediated Mitochondrial Homeostasis

doi: 10.3390/ijms27104351

Figure Lengend Snippet: The inhibition of SIRT1 abrogates the beneficial effects of cordycepin on mitochondrial function and metabolic reprogramming in brown adipocytes. All experiments were performed on cultured primary brown adipocytes. Cells were pre-treated with cordycepin (Cpn, 10 μM) and/or the SIRT1-specific inhibitor EX-527 (10 μM) for 48 h. ( A – P ) The total SIRT1 protein levels and representative Western blots for key proteins involved in PGC-1 α downstream signaling (p-ACC, TFAM, NRF1, CPT1 β , DRP1, P-DRP1, CD36) and mitophagy (PINK1, PARKIN, LC3B, p62). Corresponding densitometric quantification of the Western blot results. Tubulin and GAPDH were used as a loading control for Western blots. Data are presented as mean ± SD with individual data points shown ( n = 3 independent experiments for in vitro studies). Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Recombinant human SIRT1 (MedChemExpress, HY- P71596 , Monmouth Junction, NJ, USA) was immobilized on a CM5 sensor chip via amine coupling using the standard EDC/NHS chemistry.

Techniques: Inhibition, Cell Culture, Western Blot, Control, In Vitro

Journal: Metabolic Brain Disease

Article Title: The SIRT-1/Nrf-2/HO-1 antioxidant defense axis in adult attention-deficit/hyperactivity disorder

doi: 10.1007/s11011-026-01845-5

Figure Lengend Snippet: ROC Curve Analysis for HO-1, Nrf-2 and SIRT-1. HO-1 (cut-off value 0.87, sensitivity 78.3%, specificity 76.7%); Nrf-2 (cut-off value 9.8 sensitivity 70%, specificity 73.3%); SIRT-1 (cut-off value 8.3, sensitivity 75%, specificity 70%). Abbreviations: HO-1: heme oxygenases-1; Nrf-2: nuclear factor erythroid 2-related factor 2; SIRT-1: Sirtuin 1

Article Snippet: Serum HO-1, NRF-2, and SIRT-1 concentrations were analyzed using ELISA kits according to the manufacturers’ standard protocols (BT Lab, Human Heme Oxygenase-1: Cat. No. E0932Hu; BT Lab, Human Nuclear Factor Erythroid 2-Related Factor 2: Cat. No. E3244Hu; BT Lab, Human Sirtuin-1: Cat. No. E2557Hu; Jiaxing Korain Biotech, Jiaxing, China) on a Rel Assay automated ELISA reader (Biobase Biodusty Co., Ltd., Jinan, China).

Techniques:

Journal: Journal of Ovarian Research

Article Title: Yangjingzhongyu Decoction regulates primordial follicle initiation via LncRNA-Smad1/AMPK dual pathways in diminished ovarian reserve

doi: 10.1186/s13048-025-01955-1

Figure Lengend Snippet: Proposed mechanism of YJZYD in regulating the primordial follicle initiation rate and microenvironment. YJZYD inhibits LTCONS-00011173 expression, leading to enhanced Smad1 phosphorylation. This activates two downstream pathways: (1) AMPK/SIRT1/PGC-1α pathway promoting mitochondrial biogenesis and antioxidant function (increased SOD, ATP; decreased MDA). (2) Modulation of follicular regulatory factors (decreased PTEN, MST; increased AMH, GDF9) to balance primordial follicle initiation and growth. Solid arrows indicate direct activation; dashed arrows denote indirect regulation or material transport

Article Snippet: Protein bands were analyzed using an electrochemilluminescence (ECL) chemil uminescence kit and the bands intensity was quantified using the GIS1000 Imaging system with the following primary antibodies: Smad1 polyclonal antibody (10429-1-AP, 1:5000, Proteintech, USA), Anti-Mullerian hormone (AMH) polyclonal antibody (14461-1-AP, 1:5000, Proteintech, USA), phosphate and tensin homolog (PTEN) polyclonal antibody (10047-1-AP, 1:5000, Proteintech, USA), AMP-activated protein kinase (AMPK) polyclonal antibody (10929-2-AP, 1:5000, proteintech, Wuhan, China) and SIRT1 polyclonal antibody (A00018-1, 1:1000, Boster, Wuhan, China).

Techniques: Expressing, Phospho-proteomics, Activation Assay

Journal: Journal of Ovarian Research

Article Title: Yangjingzhongyu Decoction regulates primordial follicle initiation via LncRNA-Smad1/AMPK dual pathways in diminished ovarian reserve

doi: 10.1186/s13048-025-01955-1

Figure Lengend Snippet: Molecular docking of YJZYD monomers. A Heatmap of binding energies. B - H . 2D interaction diagrams: ( B ) AMPK-Curcumin, ( C ) AMPK-m-Cresol, ( D ) SIRT1-Apigenin, ( E ) SIRT1-Curcumin, ( F ) SIRT1-m-Cresol, ( G ) MAP3K7-Apigenin, ( H ) MAP3K7-m-Cresol. I Docking energy legend. Docking scores: Strong binding: < -8 kcal/mol; Moderate: < -5 kcal/mol

Article Snippet: Protein bands were analyzed using an electrochemilluminescence (ECL) chemil uminescence kit and the bands intensity was quantified using the GIS1000 Imaging system with the following primary antibodies: Smad1 polyclonal antibody (10429-1-AP, 1:5000, Proteintech, USA), Anti-Mullerian hormone (AMH) polyclonal antibody (14461-1-AP, 1:5000, Proteintech, USA), phosphate and tensin homolog (PTEN) polyclonal antibody (10047-1-AP, 1:5000, Proteintech, USA), AMP-activated protein kinase (AMPK) polyclonal antibody (10929-2-AP, 1:5000, proteintech, Wuhan, China) and SIRT1 polyclonal antibody (A00018-1, 1:1000, Boster, Wuhan, China).

Techniques: Binding Assay

Journal: Journal of Ovarian Research

Article Title: Yangjingzhongyu Decoction regulates primordial follicle initiation via LncRNA-Smad1/AMPK dual pathways in diminished ovarian reserve

doi: 10.1186/s13048-025-01955-1

Figure Lengend Snippet: YJZYD enhances mitochondrial function and antioxidant capacity. A AMPK/SIRT1/PGC-1α mRNA expression. B - C Corresponding protein expression and immunoblots. D - F SOD, MDA, and ATP levels. Statistical analysis ( n = 3): Model vs. Control: * p < 0.05; YJZYD vs. Model: # p < 0.05. MRNA compared to β-actin, protein compared to GAPDH

Article Snippet: Protein bands were analyzed using an electrochemilluminescence (ECL) chemil uminescence kit and the bands intensity was quantified using the GIS1000 Imaging system with the following primary antibodies: Smad1 polyclonal antibody (10429-1-AP, 1:5000, Proteintech, USA), Anti-Mullerian hormone (AMH) polyclonal antibody (14461-1-AP, 1:5000, Proteintech, USA), phosphate and tensin homolog (PTEN) polyclonal antibody (10047-1-AP, 1:5000, Proteintech, USA), AMP-activated protein kinase (AMPK) polyclonal antibody (10929-2-AP, 1:5000, proteintech, Wuhan, China) and SIRT1 polyclonal antibody (A00018-1, 1:1000, Boster, Wuhan, China).

Techniques: Expressing, Western Blot, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Electroacupuncture Pretreatment Ameliorates Perioperative Neurocognitive Disorder in Aged Mice by Inhibiting Ferroptosis Through the SIRT1 / NRF2 / GPX4 Pathway

doi: 10.1111/jcmm.71021

Figure Lengend Snippet: SIRT1/NRF2/GPX4 pathway is involved in hippocampal ferroptosis in aged mice. (A) WB images and quantification analysis of SIRT1, NRF2 and GPX4 in the hippocampus of aged mice. (B) WB images and quantification analysis of SLC7A11, TFR1, IRP2 and ferritin in the hippocampus of aged mice ( n = 3 per group). (C) qRT‐PCR expression of SIRT1, NRF2, GPX4, SLC7A11, TFR1, IRP2 and ferritin mRNA in the hippocampus of aged mice ( n = 3 per group). Values are presented as mean ± SEM. ** p < 0.01 compared with the C group; # p < 0.05 and ## p < 0.01 compared with the M group; + p < 0.05 and ++ p < 0.01 and compared with the EX group.

Article Snippet: The membrane was then incubated overnight at 4°C with primary antibodies: SIRT1 (1:850; Lot‐19G10A10; BOSTER), NRF2 (1:1500; Cat#YT3189; Immunoway), iron regulatory protein 2 (IRP2) (1:3000; Cat#YN3307; Immunoway), transferrin receptor 1 (TFR1) (1:750; LotNo‐23BP65E1; BOSTER), GPX4 (1:1500; Cat#YN3047; Immunoway), ferritin (1:3000; Cat#YT1692; Immunoway) and SLC7A11 (1:2000; Cat#YT8130; Immunoway).

Techniques: Quantitative RT-PCR, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Electroacupuncture Pretreatment Ameliorates Perioperative Neurocognitive Disorder in Aged Mice by Inhibiting Ferroptosis Through the SIRT1 / NRF2 / GPX4 Pathway

doi: 10.1111/jcmm.71021

Figure Lengend Snippet: EA ameliorates sevoflurane aesthesia‐induced ferroptosis in the hippocampus of aged mice through the SIRT1/NRF2/GPX4 signalling pathway. (A) Representative immunofluorescence staining of GPx4 (green) and NeuN (red) in the hippocampal CA1 region and fluorescence intensity of GPX4 levels in NeuN‐positive cells quantified in each group. Scale bar = 60 μm. (B) Representative TEM images of hippocampal neuron mitochondria in different groups. Scale bar = 2 μm. Values are presented as mean ± SEM ( n = 3 per group). **** p < 0.0001 compared with the C group; #### p < 0.0001 compared with the M group; ++++ p < 0.0001 compared with the EX group.

Article Snippet: The membrane was then incubated overnight at 4°C with primary antibodies: SIRT1 (1:850; Lot‐19G10A10; BOSTER), NRF2 (1:1500; Cat#YT3189; Immunoway), iron regulatory protein 2 (IRP2) (1:3000; Cat#YN3307; Immunoway), transferrin receptor 1 (TFR1) (1:750; LotNo‐23BP65E1; BOSTER), GPX4 (1:1500; Cat#YN3047; Immunoway), ferritin (1:3000; Cat#YT1692; Immunoway) and SLC7A11 (1:2000; Cat#YT8130; Immunoway).

Techniques: Immunofluorescence, Staining, Fluorescence