Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 3. VAC alleviated the renal inflammatory response and oxidative stress in T2DM mice (A) MDA contents in diabetic kidney tissues. n=6. (B) GSH-Px activity in diabetic kidney tissues. n=6. (C) Averaged fluorescence intensity of DHE fluorescence in diabetic kidney tissues. n=6. (D) Averaged fluorescence intensity of DCFH-DA fluorescence staining of diabetic kidney tissues. n=6. (E) DHE fluorescence staining or DCFH-DA fluorescence staining of diabetic kidney tissues. Scale bar: 100 μm. (F) F4/80 staining of diabetic kidney tissues. Scale bar: 50 μm, and the average fluorescence intensity of F4/80-expressing diabetic kidney tissues is shown. (G‒M) Representative blot images and quantitative analysis of phosphorylated NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. n=4. *P<0.05, **P<0.01, ***P<0.001 vs Ctrl. #P<0.05, ##P<0.01, ###P<0.001 vs DN.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Activity Assay, Fluorescence, Staining, Expressing

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 5. VAC alleviated the inflammatory response and oxidative stress in diabetic kidneys HK-2 cells were preincubated with 5 μM VAC for 12 h, followed by exposure to 35 mM HG for 48 h. (A–C) Relative mRNA levels of IL-1β, VCAM-1 and COX2. (D-F) DHE fluorescence staining or DCFH-DA fluorescence staining of HK-2 cells. Scale bar: 100 μm. (G–K) Relative mRNA levels of Nrf2, catalase, SOD3, SOD2 and SOD1. (L–R) Representative blot images and quantitative analysis of phosphorylated NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. *P<0.05, **P<0.01, ***P<0.001 vs NG. #P<0.05, ##P<0.01, ###P<0.001 vs HG. n=4.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Fluorescence, Staining

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 8. VAC ameliorated fibrosis, the inflammatory response and oxidative stress in HG-induced HK-2 cells exposed to the EGFR inhibitor AG1478 or the ERK inhibitor U0126 (A–D) Relative mRNA levels of collagen-1, TGF-β1, α-SMA and E-cadherin in HK-2 cells. (E–G) Relative mRNA levels of IL-1β, VCAM-1 and COX-2 in HK-2 cells. (H) Averaged fluorescence intensity of DHE fluorescence in HK-2 cells. (I) DHE staining was performed on HG-induced HK-2 cells. Scale bar: 100 μm. (J–P) Representative blot images and quantitative analysis of phosphorylated and total NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. *P<0.05, **P<0.01, ***P<0.001 vs NG. #P<0.05, ##P<0.01, ###P<0.001 vs HG. n=4.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Fluorescence, Staining

Journal: Cell metabolism

Article Title: Ejection of damaged mitochondria and their removal by macrophages ensure efficient thermogenesis in brown adipose tissue

doi: 10.1016/j.cmet.2022.02.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: PDHβ (MR205484L4V, Origene, Rockville, MD, USA), SOD2 (MR201642L2V, Origene, Rockville, MD, USA), PINK1 (sc-44599-V, Santa Cruz Biotechnolgies, Dallas, TX, USA) or empty vector (pLenti-C-mGFP-P2A-Puro) was inoculated at 10 multiplicities of infection (MOI) with 2 μg/mL polybrene reagent in complete growth medium and then incubated for 48 h. Cells were selected with 2 μg/mL puromycin for three passages and maintained in 1 μg/mL puromycin.

Techniques: Immunofluorescence, Recombinant, Cytometry, Purification, shRNA, Cell Culture, Lysis, XF Assay, Staining, Isolation, Transfection, Live Cell Imaging, Mouse Assay, Software, Expressing

Journal: Frontiers in Pharmacology

Article Title: Enhancing Fatty Acids Oxidation via L-Carnitine Attenuates Obesity-Related Atrial Fibrillation and Structural Remodeling by Activating AMPK Signaling and Alleviating Cardiac Lipotoxicity

doi: 10.3389/fphar.2021.771940

Figure Lengend Snippet: LCA inhibits cardiac oxidative stress and mitigates inflammation. Representative images and analysis of the subcellular localization of the oxidation products of (A,B) MitoSOX and (C,D) DHE. Red: oxidation products-staining. Scar: 10 μm n = 4. (E,F) Comparisons of serum MDA and SOD using commercially available kits among the groups. (G,H) Levels of atrial MDA and SOD normalized to tissue protein concentration. (I) Schematic diagram illustrating NRF2-related signals. (J,K) Localization of NRF2 in atria by confocal immune-cyto-chemical analysis: Blue: nucleus (DAPI); Red: NRF2-staining; Pink: merge of blue and red indicated nuclear localization of NRF2 (Arrow). Scar: 30 μm n = 4. (L,M) Representative images and quantitative analysis of anti-oxidative system involved protein expressions (NRF2 and SOD2) using Western blot. (N,O) Representative images and analysis of oxidative DNA damage by 8-OH-dG staining. Green: 8-OH-dG -staining; Blue: DAPI. Scar: 50 μm n = 4. (P,Q) Representative images and analysis of DNA damage by TUNEL staining. Red: TUNEL-staining; Blue: DAPI; Pink: merge. Scar: 50 μm n = 4. Comparisons of (R) serum LDH and serum (S) CK-MB using commercially available kits. n = 8. (T) Quantitative analysis of the expression of inflammation-related genes in the atria using RT-qPCR. (U) Representative images and quantitative analysis of expression and phosphorylation of NFκB using Western blot. One-way ANOVA with Bonferroni post-hoc test was used to compare data among groups. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01. STD, standard diet; HFD, high-fat diet; LCA, l -carnitine; DHE, dihydroethidium; MDA, molondialdehyde; SOD, superoxide dismutase; NRF2, nuclear erythroid 2 p45-related factor 2; p-, phoso-; SOD2, manganese superoxide dismutase, superoxide dismutase 2; 8-OH-Dg, 8-hydroxydeoxyguanosine; IL, interleukin; TNF-α, tumor necrosis factor-α; MCP-1, monocyte chemoattractant protein-1; NFκB, the nuclear factor kappa B; LDH, lactate dehydrogenase; CK-MB, creatine kinase-MB.

Article Snippet: The membranes were blocked with 5% skim milk and incubated with antibodies against total-AMPK (1:1,000; Cell Signaling Technology/CST, Massachusetts, United States), CD36 (1:1,000; Abcam), collagen I (1:1,000; Abcam), collagen III (1:1,000; Abcam), connexin-43 (1:200; Invitrogen), CPT1B (1:1,000; Proteintech), GLUT4 (1:500; Proteintech), NFκB (1:1,000; Proteintech), NRF2 (1:1,000; Proteintech), phoso-AMPK (Thr172; 1:1,000; CST), phoso-Akt (Ser473; 1:1,000; CST), phoso-NFκB (1:1,000; CST), PGC1α (1:1,000; Proteintech), SOD2 (1:1,000; Proteintech), TGF-β (1:1,000; Abcam), α-SMA (1:1,000; CST), β-actin (1:5,000; Proteintech).

Techniques: Staining, Protein Concentration, Western Blot, TUNEL Assay, Expressing, Quantitative RT-PCR, Phospho-proteomics

Journal: Frontiers in Pharmacology

Article Title: Enhancing Fatty Acids Oxidation via L-Carnitine Attenuates Obesity-Related Atrial Fibrillation and Structural Remodeling by Activating AMPK Signaling and Alleviating Cardiac Lipotoxicity

doi: 10.3389/fphar.2021.771940

Figure Lengend Snippet: Pharmacological inhibition of AMPK via CC attenuated LCA-conferred beneficial effects in palmitate-treated primary atrial cardiomyocytes. (A) Schematic diagram illustrating the cell isolation, culture and treatment. (B) FAO rate in different treatment groups. The measure of substrate utilization after 18°C unsaturated fatty acid (Oleate, 100 μM) addition was normalized with maximal O 2 consumption in Control cells. (C) Representative images and (D) quantitative analysis of the expression of FAO-related proteins using Western blot. (E) Cellular MDA concentrations among the groups. (F) Representative images and (G) quantitative analysis of the expression of oxidative stress-related proteins (NRFS and SOD2) and inflammation-related protein (NFκB) using Western blot. n = 3 or 4 each group. Two-way ANOVA with Bonferroni post-hoc test was used to compare data among groups. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01. CC, Compound C; LCA, l -carnitine; PA, palmitate; FAO, fatty acids oxidation; MDA, molondialdehyde; NRF2, nuclear erythroid 2 p45-related factor 2; AMPK, AMP-activated protein kinase; CD36, FAT; CPT1B, carnitine palmitoyltransferase-1B; p-, phoso-; PGC1α, peroxisome proliferator-activated receptor γ coactivator1α; NRF2, Nuclear factor erythroid 2-related factor 2; NFκB, the nuclear factor kappa B; SOD2, manganese superoxide dismutase, superoxide dismutase 2.

Article Snippet: The membranes were blocked with 5% skim milk and incubated with antibodies against total-AMPK (1:1,000; Cell Signaling Technology/CST, Massachusetts, United States), CD36 (1:1,000; Abcam), collagen I (1:1,000; Abcam), collagen III (1:1,000; Abcam), connexin-43 (1:200; Invitrogen), CPT1B (1:1,000; Proteintech), GLUT4 (1:500; Proteintech), NFκB (1:1,000; Proteintech), NRF2 (1:1,000; Proteintech), phoso-AMPK (Thr172; 1:1,000; CST), phoso-Akt (Ser473; 1:1,000; CST), phoso-NFκB (1:1,000; CST), PGC1α (1:1,000; Proteintech), SOD2 (1:1,000; Proteintech), TGF-β (1:1,000; Abcam), α-SMA (1:1,000; CST), β-actin (1:5,000; Proteintech).

Techniques: Inhibition, Cell Isolation, Control, Expressing, Western Blot

Journal: Biology of Sex Differences

Article Title: Mitochondrial function and oxidative stress in white adipose tissue in a rat model of PCOS: effect of SGLT2 inhibition

doi: 10.1186/s13293-022-00455-x

Figure Lengend Snippet: TaqMan assay IDs

Article Snippet: Mitochondrial superoxide dismutase (SOD2) , Sod2 , Rn00690588_g1.

Techniques: TaqMan Assay

Journal: Biology of Sex Differences

Article Title: Mitochondrial function and oxidative stress in white adipose tissue in a rat model of PCOS: effect of SGLT2 inhibition

doi: 10.1186/s13293-022-00455-x

Figure Lengend Snippet: Effect of EMPA on mRNA expression on antioxidant enzymes in white adipose depots in PCOS. Effect of EMPA on subcutaneous white adipose tissue (WAT) mRNA expression of A cytosolic superoxide dismutase (SOD1), B mitochondrial superoxide dismutase (SOD2), and C catalase; retroperitoneal WAT mRNA expression of D SOD1, E SOD2, and F catalase; and mesenteric WAT mRNA expression of G SOD1, H SOD2, and I catalase after 3 weeks of EMPA treatment. Expression was normalized by the geometric mean of three housekeeping genes (GMHK) and standardized to untreated control rats. Log2 values are expressed as mean ± SEM. Data were analyzed by two-way ANOVA followed by Tukey post hoc tests. Significant interactions were observed for SOD1, SOD2, and catalase in subcutaneous WAT, in SOD2 in retroperitoneal WAT, and in SOD1 in mesenteric WAT. * P < 0.05. n = 7–8 per group

Article Snippet: Mitochondrial superoxide dismutase (SOD2) , Sod2 , Rn00690588_g1.

Techniques: Expressing, Control

Journal: Biology of Sex Differences

Article Title: Mitochondrial function and oxidative stress in white adipose tissue in a rat model of PCOS: effect of SGLT2 inhibition

doi: 10.1186/s13293-022-00455-x

Figure Lengend Snippet: Effect of EMPA on protein expression on antioxidant enzymes in subcutaneous white adipose tissue in PCOS. Effect of EMPA on subcutaneous white adipose tissue (WAT) protein expression of A cytosolic superoxide dismutase (SOD1, ~ 18 kDa), B mitochondrial superoxide dismutase (SOD2, ~ 22 kDa), and C catalase (~ 60 kDa) after 3 weeks of EMPA treatment. Data were normalized by total protein content (TPC). Data are expressed as mean ± SEM and were analyzed by two-way ANOVA followed by Tukey post hoc tests. Significant interactions were observed for SOD1 and SOD2 in subcutaneous WAT. * P < 0.05. n = 3–4 per group

Article Snippet: Mitochondrial superoxide dismutase (SOD2) , Sod2 , Rn00690588_g1.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: A Preliminary Study of the Effect of Quercetin on Cytotoxicity, Apoptosis, and Stress Responses in Glioblastoma Cell Lines

doi: 10.3390/ijms23031345

Figure Lengend Snippet: The effect of QCT on oxidative stress in glioblastoma cells. A172 and LBC3 cells were incubated with 50 or 100 μmol/L of QCT for 24 h or 48 h. ROS generation in A172 ( A ) and LBC3 ( B ) cells. RT-qPCR analysis of antioxidant-enzyme genes Sod1 and Sod2 in A172 ( C ) and LBC3 ( D ) cell lines. Results shown as relative fold change in mRNA expression in comparison to untreated controls, where expression level was set as 1. Western blot analysis of SOD1 and SOD2 expressions in A172 and LBC3 cells ( E ). Mean ± SD from three independent experiments are shown. * p < 0.05.

Article Snippet: HRP-conjugated anti-rabbit IgG, polyclonal (rabbit) anti-β-tubulin, monoclonal (rabbit) anti-CHOP, monoclonal (rabbit) anti-Casp 3 (cleaved), monoclonal (rabbit) anti-Casp 9 (cleaved), polyclonal (rabbit) anti-SOD1, and monoclonal (rabbit) anti-SOD2 antibodies were provided by Cell Signaling Technology (Boston, MA, USA).

Techniques: Incubation, Quantitative RT-PCR, Expressing, Comparison, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Mitochondrial Biogenesis in Response to Chromium (VI) Toxicity in Human Liver Cells

doi: 10.3390/ijms18091877

Figure Lengend Snippet: Primers sequences used in TaqMan ® Gene Expression Assays.

Article Snippet: SOD2 , Hs00167309_m1 , 4331182.

Techniques: Gene Expression