Journal: Translational Neurodegeneration

Article Title: Deficient AMPK-SENP1-Sirt3 signaling impairs mitochondrial complex I function in Parkinson’s disease model

doi: 10.1186/s40035-025-00489-2

Figure Lengend Snippet: MPTP/MPP + attenuates AMPK activity and diminishes mitochondrial translocation of SENP1. a Relative intensities of AMPK (total lysates), P-AMPK/AMPK (total lysates), SENP1 (total lysates), SENP1 (mitochondrial lysates) and SUMO1-conjugated proteins (mitochondrial lysates) in the ventral midbrain of 3-month-old WT mice injected with MPTP or saline. Data were from 3 biological replicates, and are expressed as mean ± SD, n = 3 mice. b Left, western blotting for SENP1 (total lysates), SENP1 (mitochondrial lysates), AMPK (total lysates) and P-AMPK (total lysates) from SH-SY5Y cells treated with culture medium, MPP + , or MPP + and metformin (2 mmol/L) for 24 h, as well as blotting for SUMO1 in anti-Sirt3 immunoprecipitant from SH-SY5Y cell mitochondrial lysates. The total levels of Sirt3, SUMO1-conjugated proteins, and acetyl-conjugated proteins in mitochondrial lysates were also detected. Right, relative intensities of AMPK (total lysates), P-AMPK/AMPK (total lysates), SENP1 (total lysates), SENP1 (mitochondrial lysates), SUMOylated Sirt3 to total Sirt3 (mitochondrial lysates), SUMO1-conjugated proteins (mitochondrial lysates) and acetyl-conjugated proteins (mitochondrial lysates) from 3 biological replicates. Data expressed as mean ± SD, n = 3. c SH-SY5Y cells were treated with culture medium, MPP + , or MPP + + metformin for 24 h. CCK8 assay was performed to measure cell viability. Data expressed as mean ± SD, n = 3. One-way ANOVA with Tukey's post-hoc test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: For Western blotting assays, the following primary antibodies were used: anti-GAPDH (1:1000; Cell Signaling, 5174; Danvers, MA), anti-ATP5A (1:300; Abcam, 110273; Cambridge, MA), anti-Sirt3 (1:1000; Cell Signaling, 5490), anti-NDUFS2 (1:4000; Abcam, 110249), anti-NDUFA5 (1:1000; Proteintech, 16640; Rosemont, IL), anti-NDUFS3 (1:2000; Abcam, 14711), anti-NDUFA9 (1:1000; Abcam, 14713), anti-NDUFV1 (1:2000; Proteintech, 11238), anti-NDUFS7 (1:2000; Proteintech, 15728), anti-SENP1 (1:1000; Abcam, 108981), anti-AMP-activated protein kinase (AMPK) (1:1000; Cell Signaling, 2532), anti-SUMO1 (1:1000, custom-made by Cheng's team) [ ], anti-phosphorylated AMPK (P-AMPK) (1:1000; Cell Signaling, 2535), anti-Pan-AcK (Acetyl-conjugated proteins; 1:1000; Cell Signaling, 9441), and anti-tyrosine hydroxylase (TH)(1:200; Abcam, 112).

Techniques: Activity Assay, Translocation Assay, Injection, Saline, Western Blot, CCK-8 Assay

Journal: Frontiers in Bioscience-Landmark

Article Title: Trimetazidine: Activating AMPK Signal to Ameliorate Coronary Microcirculation Dysfunction after Myocardial Infarction

doi: 10.31083/fbl25565

Figure Lengend Snippet: Fig. 4. Trimetazidine regulates the expression of AMPK signaling pathway during myocardial I/R. The expressions of adenosine

Article Snippet: The primary antibodies used in this study included endothelial nitric oxide synthase (eNOS) (1:1000, 27120-1-AP,Wuhan, China), phosphorylated eNOS (p-eNOS) (1:1500, AF3247, Affinity, Jiangsu, China), endothelin-1 (ET-1) (1:1000, 12191- 1-AP, Proteintech, Wuhan, China), ZO-1 (1:5000, 21773- 1-AP, Proteintech, Wuhan, China), Occludin (1:1000, #91131, CST, Danvers, MA, USA), VE-cadherin (1:1000, 27956-1-AP, Proteintech, Wuhan, China), AMPK (1:2000, 10929-2AP, Proteintech, Wuhan, China), phosphorylated Adenosine Monophosphate (AMP)-activated protein kinase (p-AMPK) (1:1500, AP1002, ABclonal, Wuhan, China), kruppel-like factor 4 (KLF4) (1:1000, ab241666, Abcam, Cambridge, MA, USA), peroxisome proliferatoractivated receptor delta (PPARδ) (1:1000, 60193-1-lg, Proteintech, Wuhan, China), CD31 (1:1000, #77699, CST, Danvers, MA,USA), vascular endothelial growth factor (VEGF) (1:1500, AF5131, Affinity, Jiangsu, China), protein kinase B (AKT) (1:3000, 10176-2-AP, Proteintech, Wuhan, China), phosphorylated AKT (p-AKT) (1:2000, 66444-1-lg, Proteintech, Wuhan, China), phosphatidylinositol 3-Kinase (PI3K) (1:5000, 60225-1-lg, Proteintech, Wuhan, China), phosphorylated-PI3K (p-PI3K) (1:1000, AF4372, Affinity, Jiangsu, China) and GAPDH (1:1500, AF7021, Affinity, Jiangsu, China).

Techniques: Expressing

Journal: Frontiers in Bioscience-Landmark

Article Title: Trimetazidine: Activating AMPK Signal to Ameliorate Coronary Microcirculation Dysfunction after Myocardial Infarction

doi: 10.31083/fbl25565

Figure Lengend Snippet: Fig. 6. Effect of trimetazidine with varying concentrations on AMPK signaling pathway in primary mouse heart microvascular

Article Snippet: The primary antibodies used in this study included endothelial nitric oxide synthase (eNOS) (1:1000, 27120-1-AP,Wuhan, China), phosphorylated eNOS (p-eNOS) (1:1500, AF3247, Affinity, Jiangsu, China), endothelin-1 (ET-1) (1:1000, 12191- 1-AP, Proteintech, Wuhan, China), ZO-1 (1:5000, 21773- 1-AP, Proteintech, Wuhan, China), Occludin (1:1000, #91131, CST, Danvers, MA, USA), VE-cadherin (1:1000, 27956-1-AP, Proteintech, Wuhan, China), AMPK (1:2000, 10929-2AP, Proteintech, Wuhan, China), phosphorylated Adenosine Monophosphate (AMP)-activated protein kinase (p-AMPK) (1:1500, AP1002, ABclonal, Wuhan, China), kruppel-like factor 4 (KLF4) (1:1000, ab241666, Abcam, Cambridge, MA, USA), peroxisome proliferatoractivated receptor delta (PPARδ) (1:1000, 60193-1-lg, Proteintech, Wuhan, China), CD31 (1:1000, #77699, CST, Danvers, MA,USA), vascular endothelial growth factor (VEGF) (1:1500, AF5131, Affinity, Jiangsu, China), protein kinase B (AKT) (1:3000, 10176-2-AP, Proteintech, Wuhan, China), phosphorylated AKT (p-AKT) (1:2000, 66444-1-lg, Proteintech, Wuhan, China), phosphatidylinositol 3-Kinase (PI3K) (1:5000, 60225-1-lg, Proteintech, Wuhan, China), phosphorylated-PI3K (p-PI3K) (1:1000, AF4372, Affinity, Jiangsu, China) and GAPDH (1:1500, AF7021, Affinity, Jiangsu, China).

Techniques:

Journal: Frontiers in Bioscience-Landmark

Article Title: Trimetazidine: Activating AMPK Signal to Ameliorate Coronary Microcirculation Dysfunction after Myocardial Infarction

doi: 10.31083/fbl25565

Figure Lengend Snippet: Fig. 8. Trimetazidine alleviates functional impairment of OGD/R-induced primary mouse heart microvascular endothelial cells through AMPK signaling pathway. (A) The expressions of eNOS, p-eNOS and ET-1 were detected by western blot. (B) The NO

Article Snippet: The primary antibodies used in this study included endothelial nitric oxide synthase (eNOS) (1:1000, 27120-1-AP,Wuhan, China), phosphorylated eNOS (p-eNOS) (1:1500, AF3247, Affinity, Jiangsu, China), endothelin-1 (ET-1) (1:1000, 12191- 1-AP, Proteintech, Wuhan, China), ZO-1 (1:5000, 21773- 1-AP, Proteintech, Wuhan, China), Occludin (1:1000, #91131, CST, Danvers, MA, USA), VE-cadherin (1:1000, 27956-1-AP, Proteintech, Wuhan, China), AMPK (1:2000, 10929-2AP, Proteintech, Wuhan, China), phosphorylated Adenosine Monophosphate (AMP)-activated protein kinase (p-AMPK) (1:1500, AP1002, ABclonal, Wuhan, China), kruppel-like factor 4 (KLF4) (1:1000, ab241666, Abcam, Cambridge, MA, USA), peroxisome proliferatoractivated receptor delta (PPARδ) (1:1000, 60193-1-lg, Proteintech, Wuhan, China), CD31 (1:1000, #77699, CST, Danvers, MA,USA), vascular endothelial growth factor (VEGF) (1:1500, AF5131, Affinity, Jiangsu, China), protein kinase B (AKT) (1:3000, 10176-2-AP, Proteintech, Wuhan, China), phosphorylated AKT (p-AKT) (1:2000, 66444-1-lg, Proteintech, Wuhan, China), phosphatidylinositol 3-Kinase (PI3K) (1:5000, 60225-1-lg, Proteintech, Wuhan, China), phosphorylated-PI3K (p-PI3K) (1:1000, AF4372, Affinity, Jiangsu, China) and GAPDH (1:1500, AF7021, Affinity, Jiangsu, China).

Techniques: Functional Assay, Western Blot

Journal: Frontiers in Bioscience-Landmark

Article Title: Trimetazidine: Activating AMPK Signal to Ameliorate Coronary Microcirculation Dysfunction after Myocardial Infarction

doi: 10.31083/fbl25565

Figure Lengend Snippet: Fig. 9. Trimetazidine alleviates barrier damage of OGD/R-induced primary mouse heart microvascular endothelial cells through AMPK signaling pathway. (A) The endothelial cell permeability experiment was used to detect the cell permeability (scale bar: 50

Article Snippet: The primary antibodies used in this study included endothelial nitric oxide synthase (eNOS) (1:1000, 27120-1-AP,Wuhan, China), phosphorylated eNOS (p-eNOS) (1:1500, AF3247, Affinity, Jiangsu, China), endothelin-1 (ET-1) (1:1000, 12191- 1-AP, Proteintech, Wuhan, China), ZO-1 (1:5000, 21773- 1-AP, Proteintech, Wuhan, China), Occludin (1:1000, #91131, CST, Danvers, MA, USA), VE-cadherin (1:1000, 27956-1-AP, Proteintech, Wuhan, China), AMPK (1:2000, 10929-2AP, Proteintech, Wuhan, China), phosphorylated Adenosine Monophosphate (AMP)-activated protein kinase (p-AMPK) (1:1500, AP1002, ABclonal, Wuhan, China), kruppel-like factor 4 (KLF4) (1:1000, ab241666, Abcam, Cambridge, MA, USA), peroxisome proliferatoractivated receptor delta (PPARδ) (1:1000, 60193-1-lg, Proteintech, Wuhan, China), CD31 (1:1000, #77699, CST, Danvers, MA,USA), vascular endothelial growth factor (VEGF) (1:1500, AF5131, Affinity, Jiangsu, China), protein kinase B (AKT) (1:3000, 10176-2-AP, Proteintech, Wuhan, China), phosphorylated AKT (p-AKT) (1:2000, 66444-1-lg, Proteintech, Wuhan, China), phosphatidylinositol 3-Kinase (PI3K) (1:5000, 60225-1-lg, Proteintech, Wuhan, China), phosphorylated-PI3K (p-PI3K) (1:1000, AF4372, Affinity, Jiangsu, China) and GAPDH (1:1500, AF7021, Affinity, Jiangsu, China).

Techniques: Permeability

Journal: Diabetes & Metabolism Journal

Article Title: Shionone Inhibits Glomerular Fibirosis by Suppressing NLRP3 Related Inflammasome though SESN2-NRF2/HO-1 Pathway

doi: 10.4093/dmj.2024.0024

Figure Lengend Snippet: Shionone (SH) improved glomerulus fibrosis and inflammatory in diabetic mice. (A) The histopathology analysis of glomeruli using Masson staining, ×200. (B) Quantitative analysis of Masson staining. (C) The histopathology analysis of glomeruli using Periodic acid–Schiff (PAS) staining, ×200. (D) Quantitative analysis of PAS staining. (E) Subcellular structure analysis of podocytes detected by transmission electron microscopy, ×10,000. (F) Expression of α-smooth muscle actin (α-SMA), NLR family pyrin domain containing 3 (NLRP3), interleukin 1β (IL-1β), phosphorylated adenosine monophosphate-activated protein kinase (P-AMPK) in glomeruli through immunohistochemistry, ×200. (G) α-SMA, NLRP3, IL-1β, P-AMPK proteins from wild-type mice detected by Western blot. (H) Quantitative analysis of Western blot. Data are expressed as the mean±standard error of the mean. a P <0.05, b P <0.01, c P <0.001, compared with the control group; d P <0.05, e P <0.01, f P <0.001 compared with the type 2 diabetes mellitus (T2DM) group; g P <0.05, compared with the irbesartan group.

Article Snippet: The sections were incubated with SESN2, α-smooth muscle actin (α-SMA), NLPR3, and phosphorylated AMPK (P-AMPK) antibodies obtained from Santa Cruz Biotechnology (Dallas, TX, USA) at a dilution of 1:200.

Techniques: Histopathology, Staining, Transmission Assay, Electron Microscopy, Expressing, Immunohistochemistry, Western Blot, Control

Journal: Diabetes & Metabolism Journal

Article Title: Shionone Inhibits Glomerular Fibirosis by Suppressing NLRP3 Related Inflammasome though SESN2-NRF2/HO-1 Pathway

doi: 10.4093/dmj.2024.0024

Figure Lengend Snippet: Shionone (SH)’s anti-fibrosis effect was sestrin-2 (SESN2) dependent in diabetic mice. (A) The histopathology analysis of SESN2 -/- glomeruli using Masson staining, ×200. (B) Quantitative analysis of Masson staining. (C) The histopathology analysis of SESN2 -/- glomeruli using Periodic acid–Schiff (PAS) staining, ×200. (D) Quantitative analysis of PAS staining. (E) Subcellular structure analysis of podocytes in SESN2 -/- mice detected by transmission electron microscopy, ×10,000. (F) Expression of SESN2, phosphorylated adenosine monophosphate-activated protein kinase (PAMPK), NLR family pyrin domain containing 3 (NLRP3), interleukin 1β (IL-1β), α-smooth muscle actin (α-SMA) in SESN2 -/- glomeruli through immunohistochemistry, ×200. (G) P-AMPK, NLRP3, IL-1β, α-SMA proteins from SESN2 -/- mice detected by Western blot. (H) Quantitative analysis of Western blot. (I) SESN2, nuclear factor erythroid 2-related factor 2 (NRF2), heme oxygenase- 1 (HO-1) proteins from SESN2 -/- mice detected by Western blot. (J) Effect of SH on the nucleus expression of NRF2 in type 2 diabetes mellitus (T2DM) SESN2 -/- mice kidney was detected by Western blot. (K) Quantitative analysis of Western blot. (L) SESN2, NRF2 in detected by confocal scanning microscopy in SESN2 -/- mice, ×200, ×500. The position indicated by the white arrows is the site of nuclear import of NRF2. Data are expressed as the mean±standard error of the mean. DAPI, 4´,6-diamidino-2-phenylindole. a P <0.05, b P <0.01, c P <0.001, compared with the control group; d P <0.05, e P <0.01, f P <0.001 compared with the SESN2 -/- group.

Article Snippet: The sections were incubated with SESN2, α-smooth muscle actin (α-SMA), NLPR3, and phosphorylated AMPK (P-AMPK) antibodies obtained from Santa Cruz Biotechnology (Dallas, TX, USA) at a dilution of 1:200.

Techniques: Histopathology, Staining, Transmission Assay, Electron Microscopy, Expressing, Immunohistochemistry, Western Blot, Microscopy, Control

Journal: Diabetes & Metabolism Journal

Article Title: Shionone Inhibits Glomerular Fibirosis by Suppressing NLRP3 Related Inflammasome though SESN2-NRF2/HO-1 Pathway

doi: 10.4093/dmj.2024.0024

Figure Lengend Snippet: Shionone (SH) could inhibit NLR family pyrin domain containing 3 (NLRP3) via sestrin-2 (SESN2)-nuclear factor erythroid 2-related factor 2 (NRF2)/heme oxygenase-1 (HO-1) in high glucose (HG) treated mouse podocyte clone 5 (MPC-5) cells. (A) Expression of SESN2, phosphorylated adenosine monophosphate-activated protein kinase (P-AMPK), NRF2, HO-1, NLRP3, interleukin 1β (IL-1β), α-smooth muscle actin (α-SMA) in MPC-5 cells detected by Western blot. (B) Quantitative analysis of Western blot. (C) Effect of SH on the mRNA level of SESN2, NRF2, NLRP3, α-SMA in MPC-5 cells. Quantitative analysis of Western blot. Data are expressed as the mean±standard error of the mean. a P <0.05, b P <0.01, c P <0.001, compared with the control group; d P <0.05, e P <0.01, f P <0.001 compared with the SESN2 -/- group; g P <0.05, h P <0.01, i P <0.001 compared with the SESN2 +/+ group.

Article Snippet: The sections were incubated with SESN2, α-smooth muscle actin (α-SMA), NLPR3, and phosphorylated AMPK (P-AMPK) antibodies obtained from Santa Cruz Biotechnology (Dallas, TX, USA) at a dilution of 1:200.

Techniques: Expressing, Western Blot, Control