Journal: Arthritis Research & Therapy

Article Title: Sirt3 improves monosodium urate crystal-induced inflammation by suppressing Acod1 expression

doi: 10.1186/s13075-023-03107-6

Figure Lengend Snippet: MSU crystals stimulation reduced the expression and deacetylation activity of Sirt3. a Sirt3 mRNA and protein levels were detected in PBMCs of health controls (HC), patients with acute gout (AG), and intermittent gout (IG). Sirt3 mRNA detection (HC, n = 24), (AG, n = 24), (IG, n = 24); GAPDH was used as an internal reference. Sirt3 protein detection (HC, n = 6), (AG, n = 6), (IG, n = 6), TUBULIN as an internal reference. b BMDMs were respectively stimulated with C16:0, MSU, or C16:0 + MSU for 12 h and Western blot was used to detect Sirt3 protein expression. c The protein levels of Sirt3 in the mouse paw treated with MSU crystals or vehicle. Data presented as mean ± SD ( n = 5 mice per group). * P < 0.05. NS means no statistical difference. d Sirt3 agonists (Viniferin) inhibited the acetylation level of mitochondrial proteins in BMDMs treated with Viniferin (1 μM) and C16:0 + MSU crystals for 12 h. Acetyl-conjugated proteins in mitochondrial proteins were blotted with acetylated-lysine antibody. e Viniferin treatment decreased K68 lysine acetylation of mitochondrial SOD2 protein and mitochondrial proteins were analyzed by Western blot in BMDMs. f The activity of SOD in the mitochondrial proteins of BMDMs was detected with WST-8 method. Data are mean ± SD. Three independent repetitive experiments were conducted for each result. *P < 0.05. NS represents P > 0.05

Article Snippet: HY-133987, Mito-TEMPO, and Viniferin were purchased from MedChemExpress (MCE).

Techniques: Expressing, Activity Assay, Western Blot

Journal: Journal of Molecular Endocrinology

Article Title: Uncoupling protein-2 increases nitric oxide production and TNFAIP3 pathway activation in pancreatic islets

doi: 10.1530/JME-10-0117

Figure Lengend Snippet: Characterization of basal NF-κB pathway and apoptosis in islets from UCP2 +/+ and UCP2 −/− mice. (A) Basal NF-κB (RelA subunit) activation ( n =4) in control cells and those incubated with IKKβ inhibitor BAY 11-7082 (10 μM) or iNOS inhibitor 1400W (10 μM) for 2 h. Data were normalized to protein concentration. (B) Basal IκBα phosphorylation in the presence or absence of BAY ( n =4). Effects were quantified by chemiluminescence and normalized to protein concentration. (C) Basal apoptosis, as a percentage of total β-cells, in UCP2 +/+ and UCP2 −/− islets, comparing the effects of BAY or 1400W with untreated controls. Cell apoptosis was assessed by TUNEL staining. Two thousand cells were counted on average at 200× magnification ( n =4). * P <0·05, effect of inhibitor; § P <0·05, effect of genotype).

Article Snippet: The active NF-κB contained in the nuclear extracts was measured by its DNA-binding activity on immobilized oligonucleotides encoding a specific consensus site using a NF-κB RelA transcription factor ELISA kit (Active Motif).

Techniques: Activation Assay, Control, Incubation, Protein Concentration, Phospho-proteomics, TUNEL Assay, Staining

Journal: Antioxidants

Article Title: Oleanolic Acid Nanofibers Attenuated Particulate Matter-Induced Oxidative Stress in Keratinocytes

doi: 10.3390/antiox10091411

Figure Lengend Snippet: Oleanolic acid inhibits protein expression related to inflammation and aging in PM−induced keratinocytes damage through the ROS/MAPKs pathway. ( A ) Cell viability, ( B ) ROS production, ( C ) COX-2 and NF-κB, ( D ) MMP and TIMP protein expression, and ( E ) phosphorylation of MAPKs. # represents p < 0.05 when compared with negative control. * represents p < 0.05 when compared with PM-induced control.

Article Snippet: The primary antibodies used in this study included cyclooxygenase-2 (COX-2), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metallopro-teinase-1(TIMP-1), stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK) (Cell Signaling Technology, Danvers, MA, USA), GAPDH (Santa Cruz Bi-otechnology, Dallas, TX, USA), matrix metalloproteinase-1 (MMP-1) (Proteintech Group, Rosemont, IL, USA), p38α, extracellular regulated protein kinases (ERK), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) (Merck Millipore, Burlington, MA, USA).

Techniques: Expressing, Negative Control

Journal: Antioxidants

Article Title: Oleanolic Acid Nanofibers Attenuated Particulate Matter-Induced Oxidative Stress in Keratinocytes

doi: 10.3390/antiox10091411

Figure Lengend Snippet: Oleanolic acid nanofiber inhibits the protein expression of inflammation and aging in PM−induced keratinocytes damage through the ROS/MAPKs pathway. ( A ) Cell viability, ( B ) ROS production, ( C ) COX-2 and NF-κb, ( D ) MMPs and TIMP-1 protein expression, ( E ) phosphorylation of MAPKs. A p -value < 0.05 was interpreted as statistically significant, # refers to PM treatment versus negative control, * refers to PM treatment versus OAnf, and $ refers to OA/PBS versus OAnf.

Article Snippet: The primary antibodies used in this study included cyclooxygenase-2 (COX-2), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metallopro-teinase-1(TIMP-1), stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK) (Cell Signaling Technology, Danvers, MA, USA), GAPDH (Santa Cruz Bi-otechnology, Dallas, TX, USA), matrix metalloproteinase-1 (MMP-1) (Proteintech Group, Rosemont, IL, USA), p38α, extracellular regulated protein kinases (ERK), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) (Merck Millipore, Burlington, MA, USA).

Techniques: Expressing, Negative Control