Journal: International Journal of Molecular Sciences

Article Title: Contemporary Antiretroviral Therapy Dysregulates Iron Transport and Augments Mitochondrial Dysfunction in HIV-Infected Human Microglia and Neural-Lineage Cells

doi: 10.3390/ijms241512242

Figure Lengend Snippet: ( a – l ) Expression of NDUFS1, SDHB, Nox2, and UQCRFS1 in uninfected, latently infected HIV+(GFP−), and activated HIV+(GFP+) hμglia after exposure to Regimens1 and 2 for 24 h. ( m ) Immunoblot images of protein expression in hμglia and ( n – q ) SH-SY5Y cells. Abbreviations: D− , no drug (vehicle control); R1 , Regimen1; R2, Regimen2. NDUFS1, SDHB, and UQCRFS1 are mitochondrial respiratory subunit proteins; Sod2 , a mitochondrial ROS-scavenging enzyme; Nox2 , NADPH oxidase 2, a major cytoplasmic oxidase; Tom20 , a mitochondrial protein translocase; Lrfn2 , a marker of neuronal/synaptic integrity; I + III 2 and III 2 + I V, mitochondrial supercomplexes. * p < 0.05; ** p < 0.005 (Mann–Whitney U test).

Article Snippet: Other antibodies used were: NADPH oxidase 2 (Nox2 rabbit polyclonal antibody, 1:500, 19013-1-AP, Proteintech, Chicago, IL, USA), mitochondrial superoxide dismutase 2, (Sod2, rabbit polyclonal antibody, 1:500, 24127-1, Proteintech, USA), NADH: ubiquinone oxidoreductase core subunit S1 (NDUFS1, rabbit polyclonal antibody, 1:500, 12444-1-AP, Proteintech, USA), succinate dehydrogenase complex iron-sulfur subunit B (SDHB, rabbit polyclonal antibody, 1:500, 10620-1-AP, Proteintech, USA), mitoferrin (rabbit polyclonal antibody, 1:500, 26469-1-AP Proteintech, USA), Tom20 antibody (rabbit monoclonal antibody, 1:500, 42,406, Cell Signaling Technology, Danvers, MA, USA), CD71/transferrin receptor (TfR, rabbit monoclonal antibody, 1:500, 13113, Cell Signaling Technology, USA), and tubulin (rabbit monoclonal antibody, 1:500, 9099, Cell Signaling Technology, USA) overnight at 4 °C.

Techniques: Expressing, Infection, Western Blot, Marker, MANN-WHITNEY

Journal: Disease Markers

Article Title: Pyrrosia petiolosa Extract Ameliorates Ethylene Glycol-Induced Urolithiasis in Rats by Inhibiting Oxidative Stress and Inflammatory Response

doi: 10.1155/2022/1913067

Figure Lengend Snippet: Effect of Pyrrosia petiolosa extract on oxidative stress in renal tissue of ethylene glycol-induced urolithiasis in rats. (a–c) Biochemical detection for the level of SOD, GSH, and MDA in the kidneys of rats in each group, respectively. (d) Western blot detection of NOX2 and NOX4 protein expression level in the kidney tissues of rats in each group ( n = 3). ∗∗ P < 0.01 vs. control group; # P < 0.05, ## P < 0.01 vs. EG group.

Article Snippet: Then, 5% skimmed milk was added for blocking 1 h. Next, antibodies NADPH oxidases 2 (NOX2), NOX4, TGF- β 1, p-Smad3, Smad3, p-Smad2, and Smad2 (Proteintech, Chicago, USA) were added, respectively, and the incubation was performed overnight at 4°C in a shaker.

Techniques: Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Losartan Ameliorates Calcium Oxalate-Induced Elevation of Stone-Related Proteins in Renal Tubular Cells by Inhibiting NADPH Oxidase and Oxidative Stress

doi: 10.1155/2018/1271864

Figure Lengend Snippet: ROS and NADPH oxidase were upregulated under stimulation of CaOx crystals. (a) NRK-52E cells were induced by COM (1 mM) for 6 h with or without apocynin preincubation, and then intracellular ROS was detected by flow cytometry. Expression of Nox2 and Nox4 was upregulated in CaOx-induced NRK-52E cells (b) and rat kidneys in the hyperoxaluria group (c), as determined by Western blotting analysis, and the alteration could be reversed by apocynin or losartan administration. (d) COM (1 mM) reduced cellular SOD and CAT activities and increased MDA and 8-OHdG expression in NRK-52E cells which was reversed by apocynin administration. The data are expressed as mean ± SEM. ∗ P < 0.05 compared with the Ang II group or control group and # P < 0.05 compared with the COM + Ang II group or hyperoxaluria group.

Article Snippet: Anti-AT1R, anti-NADPH oxidase 2 (Nox2), anti-p50, and anti-p65 antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Flow Cytometry, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Losartan Ameliorates Calcium Oxalate-Induced Elevation of Stone-Related Proteins in Renal Tubular Cells by Inhibiting NADPH Oxidase and Oxidative Stress

doi: 10.1155/2018/1271864

Figure Lengend Snippet: CaOx induced ROS generation and overproduction of stone-related proteins was activated via Ang II/AT1R. (a) The mRNA level of AT1R in NRK-52E cells transfected with AT1R siRNA-1, siRNA-2, or siRNA-3 compared with the negative control group (NC group) (scrambled siRNA transfected) and the mock group (transfection reagent treated only). ∗ P < 0.05 compared with the NC group and # P < 0.05 compared with the mock group. (b) The protein level of AT1R was downregulated in the COM + Ang II + AT1R siRNA-3 group compared with the COM + Ang II group. (c) The ROS generation was decreased in the COM + Ang II + AT1R siRNA-3 group compared with the COM + Ang II group. The expression of NADPH oxidase subunits (Nox2 and Nox4) (d), NF- κ B subunits (p50 and p65) (e), and stone-related proteins (OPN, CD44, and MCP-1) (f) in the COM + Ang II and COM + Ang II + AT1R siRNA-3 groups was detected by Western blotting in NRK-52E cells. The data are expressed as mean ± SEM. ∗ P < 0.05 compared with the Ang II group and # P < 0.05 compared with the COM + Ang II group. ∗# P < 0.05 compared with the Ang II group and the COM + Ang II group.

Article Snippet: Anti-AT1R, anti-NADPH oxidase 2 (Nox2), anti-p50, and anti-p65 antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Transfection, Negative Control, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Losartan Ameliorates Calcium Oxalate-Induced Elevation of Stone-Related Proteins in Renal Tubular Cells by Inhibiting NADPH Oxidase and Oxidative Stress

doi: 10.1155/2018/1271864

Figure Lengend Snippet: Losartan downregulated NF- κ B pathway activity and stone-related protein expression by attenuation of renal tubular cell oxidative stress. (a) NRK-52E cells were induced by COM (1 mM) for 6 h with or without losartan preincubation, and then intracellular ROS was detected by flow cytometry. The expression of NADPH oxidase subunits (Nox2 and Nox4) (b), NF- κ B subunits (p50 and p65) (c), and stone-related proteins (OPN, CD44, and MCP-1) (d) was decreased in the COM + Ang II + losartan group compared with the COM + Ang II group in NRK-52E cells. (e) Losartan administration ameliorated the CaOx depositions (black arrow) compared with the rat kidneys of the hyperoxaluria group (Von Kossa staining, magnification: ×400). The data are expressed as mean ± SEM. ∗ P < 0.05 compared with the Ang II group and # P < 0.05 compared with the COM + Ang II group. ∗# P < 0.05 compared with the Ang II group and the COM + Ang II group.

Article Snippet: Anti-AT1R, anti-NADPH oxidase 2 (Nox2), anti-p50, and anti-p65 antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Activity Assay, Expressing, Flow Cytometry, Staining

Journal: BioMed Research International

Article Title: Schisandrin B Diet Inhibits Oxidative Stress to Reduce Ferroptosis and Lipid Peroxidation to Prevent Pirarubicin-Induced Hepatotoxicity

doi: 10.1155/2022/5623555

Figure Lengend Snippet: SchB diet inhibited ferroptosis and oxidative stress on liver of THP-induced hepatotoxicity rats. The levels of total Fe (a), Fe 2+ (b), and Fe 3+ (c) decreased in the liver of THP rats. The levels of NRF2, SOD2, GPX4, and Bcl-2/Bax decreased in the liver of THP rats (d). In addition, the expression of NOX2/4 and cleaved caspase-3 was slightly increased (d). However, SchB diet eliminated these adverse reactions (a–d). Semiquantitative analysis provides more evidence (e). Values are expressed as mean ± SEM. ∗ P < 0.05 vs. CON and # P < 0.05 vs. THP. CON: control; SchB: schisandrin B; THP: pirarubicin; NRF2: nuclear factor erythroid 2 like 2; NOX2/4: NADPH oxidase 2/4; SOD2: superoxide disproportionation 2; GPX4: glutathione peroxidase 4; Bcl-2: B-cell lymphoma-2; Bax: Bcl-2-associated X.

Article Snippet: Antibodies recognizing nuclear factor erythroid 2 like 2 (NRF2, Cat No. 16396-1-AP), NADPH oxidase 2/4 (NOX2/4, Cat No. 19013-1-AP/14347-1-AP), superoxide disproportionation 2 (SOD2, Cat No. 24127-1-AP), glutathione peroxidase 4 (GPX4, Cat No. 14432-1-AP), B-cell lymphoma-2 (Bcl-2, Cat No. 12789-1-AP), Bcl-2-associated X (Bax, Cat No. 50599-2-Ig), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Cat No. 10494-1-AP) were bought from Proteintech (Wuhan, China).

Techniques: Expressing

Journal: BioMed Research International

Article Title: Schisandrin B Diet Inhibits Oxidative Stress to Reduce Ferroptosis and Lipid Peroxidation to Prevent Pirarubicin-Induced Hepatotoxicity

doi: 10.1155/2022/5623555

Figure Lengend Snippet: SchB inhibited ferroptosis and oxidative stress on hepatocytes cultured with THP. Effects of different concentrations of THP on cell viability (a). Effects of different concentrations of SchB with 5 μ M THP on cell viability (b). The levels of ROS increased, which decreased with Fer-1 or SchB treatment in THP hepatocytes (c). The relative fluorescence intensity provides more evidence (d). The levels of NRF2, SOD2, GPX4, and Bcl-2/Bax decreased in THP hepatocytes (e). In addition, the expression of NOX2/4 and cleaved caspase-3 was slightly increased (e). However, SchB treatment eliminated these adverse reactions (e). Semiquantitative analysis provides more evidence (f). Values are expressed as mean ± SEM. ∗ P < 0.05 vs. CON and # P < 0.05 and & P < 0.05 vs. THP. CON: control; SchB: schisandrin B; THP: pirarubicin; Fer-1: ferrostatin-1; NRF2: nuclear factor erythroid 2 like 2; NOX2/4: NADPH oxidase 2/4; SOD2: superoxide disproportionation 2; GPX4: glutathione peroxidase 4; Bcl-2: B-cell lymphoma-2; Bax: Bcl-2-associated X.

Article Snippet: Antibodies recognizing nuclear factor erythroid 2 like 2 (NRF2, Cat No. 16396-1-AP), NADPH oxidase 2/4 (NOX2/4, Cat No. 19013-1-AP/14347-1-AP), superoxide disproportionation 2 (SOD2, Cat No. 24127-1-AP), glutathione peroxidase 4 (GPX4, Cat No. 14432-1-AP), B-cell lymphoma-2 (Bcl-2, Cat No. 12789-1-AP), Bcl-2-associated X (Bax, Cat No. 50599-2-Ig), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Cat No. 10494-1-AP) were bought from Proteintech (Wuhan, China).

Techniques: Cell Culture, Fluorescence, Expressing