In IL-1β-challenged human placental explants, 200 μM resveratrol for 20 h reduced several inflammatory transcripts and secreted mediators. MCP-1 mRNA reduction was nonsignificant (P = 0.07). This was an ex vivo model using tissue from normal-glucose-tolerant pregnancies. [3]
In TNF-α-challenged human skeletal-muscle explants, resveratrol co-treatment improved insulin-signalling readouts and insulin-stimulated glucose uptake. These findings describe a tissue model and do not establish prevention or treatment of gestational diabetes. [3]
Substrate-dependent SIRT1 assay response
Biochemical assay · SIRT1 deacetylation of TAMRA-p53 and unlabeled native p53 peptides [1]
HPLC product quantification showed activation with the labeled peptide, but no activation of the native p53 peptide at 30 or 100 μM; EX-527 inhibited the native-peptide reaction.
Literature-Reported Treatment Conditions
- Treatment concentration
- Labeled peptide: resveratrol 0.01–600 μM; native peptide: 30 and 100 μM
- Exposure duration
- 30 min at room temperature
- Method
- HPLC; 1 μM peptide, 150 μM NAD+; 5 nM SIRT1 (TAMRA) or 2.5 nM (native)
- Vehicle
- 2% DMSO
- Co-treatment
- None; other compounds were separate comparators.
- Reported limits
- Numerical replicate count not established in the selected text; plotted error bars are SD.
Methods and source locations
Compound reported in the paper: Resveratrol; Compound label in the paper: resveratrol
Reported resveratrol experiments; stereochemical purity and equivalence to A10785 are not established.
Experimental Procedures: TAMRA-p53 and native-p53 HPLC assays; Results and Fig. 2C, Fig. 3C
TAMRA-p53 deacetylation: Concentration giving 50% increased enzyme activity; not EC50 or IC50. — Experimental Procedures: TAMRA-p53 and native-p53 HPLC assays; Results and Fig. 2C, Fig. 3C
Native p53 peptide deacetylation: HPLC; Fig. 3C, SD error bars; EX-527 100 μM inhibitory control. — Experimental Procedures: TAMRA-p53 and native-p53 HPLC assays; Results and Fig. 2C, Fig. 3C
Fluorophore-free peptide validation of SIRT1 modulation
Biochemical assay · Full-length human Sirt1 with SF38A-K23, DDX3X-K118 and histone H3-K116 peptides [2]
At 100 μM resveratrol, SF38A peptide deacetylation increased, DDX3X was unchanged, and H3 was inhibited. SF38A titration yielded a substrate-specific EC50.
Literature-Reported Treatment Conditions
- Treatment concentration
- 100 μM comparison; concentration titration for SF38A
- Exposure duration
- Time series; product levels after 30 min or fitted deacetylation rates
- Method
- LC-MS; comparison: 50 μM NAD+, 5 μM peptide, 25–200 nM Sirt1 at 37°C. Titration: 500 μM NAD+ at 25°C.
- Vehicle
- 1% (v/v) DMSO maintained in controls and titration
- Co-treatment
- None
- Reported limits
- At least three repetitions; displayed data are representative. Meaning of ± in reported EC50 not specified. Exact concentration assigned to each enzyme/peptide comparison not separately given.
Methods and source locations
Compound reported in the paper: Resveratrol; Compound label in the paper: resveratrol
Reported resveratrol experiments; stereochemical purity and equivalence to A10785 are not established.
Main narrative paragraph on solution validation; Fig. 1B–C; Materials and Methods: Chemicals and Mass spectrometry deacetylation assays
Peptide deacetylation response: LC-MS comparison with no enzyme, enzyme alone, and enzyme plus resveratrol in Fig. 1B. — Main narrative paragraph on solution validation; Fig. 1B–C; Materials and Methods: Chemicals and Mass spectrometry deacetylation assays
SF38A-K23 peptide deacetylation activation: Text-reported fit; Fig. 1C. ± retained without assigning SD or SEM. — Main narrative paragraph on solution validation; Fig. 1B–C; Materials and Methods: Chemicals and Mass spectrometry deacetylation assays
IL-1β-challenged human placental explants
Cell-based study · Human term placenta from normal-glucose-tolerant women undergoing elective Caesarean section; n = 6 donors [3]
Resveratrol reduced IL-1α, IL-6 and IL-8 mRNA and IL-6, IL-8 and MCP-1 release versus IL-1β alone. MCP-1 mRNA reduction did not reach significance.
Literature-Reported Treatment Conditions
- Treatment concentration
- 200 μM resveratrol; 5 ng/ml IL-1β
- Exposure duration
- 20 h
- Method
- Ex vivo co-incubation; qRT-PCR normalized to B2M and 18S rRNA; ELISA corrected for total protein
- Co-treatment
- IL-1β challenge; basal and IL-1β-only controls
- Reported limits
- Vehicle and a resveratrol-only control are not specified for this experiment. Tissue viability control not located in acquired main text.
Methods and source locations
Compound reported in the paper: Resveratrol; Compound label in the paper: resveratrol
Reported resveratrol experiments; stereochemical purity and equivalence to A10785 are not established.
Materials and methods: Tissue collection, Tissue explants, qRT-PCR, Enzyme immunoassays, Statistical analysis; Results: placenta; Fig. 2A–G
Inflammatory mRNA and mediator release: n = 6 patients; duplicate experiments averaged per donor; mean ± SEM; Fig. 2A–C,E–G. — Materials and methods: Tissue collection, Tissue explants, qRT-PCR, Enzyme immunoassays, Statistical analysis; Results: placenta; Fig. 2A–G
MCP-1 mRNA: qRT-PCR; nonsignificant decrease, Fig. 2D. — Materials and methods: Tissue collection, Tissue explants, qRT-PCR, Enzyme immunoassays, Statistical analysis; Results: placenta; Fig. 2A–G
TNF-α-challenged human skeletal-muscle explants
Cell-based study · Rectus pyramidalis explants from term elective Caesarean donors with normal glucose tolerance; n = 6 patients [3]
Resveratrol co-treatment countered the TNF-α-associated decreases in insulin-signalling readouts and insulin-stimulated 2-deoxyglucose uptake.
Literature-Reported Treatment Conditions
- Treatment concentration
- 200 μM resveratrol; 10 ng/ml TNF-α
- Exposure duration
- 20 h co-incubation, then 0.1 μM insulin for 30 min
- Method
- Western blot pIR-β/IR-β, pIRS-1/IRS-1 and GLUT4/β-actin; radiolabeled 2DG uptake with cytochalasin-B subtraction
- Co-treatment
- TNF-α challenge followed by insulin stimulation; basal and TNF-α-only controls
- Reported limits
- Vehicle and resveratrol-only control are not specified; no inference of direct SIRT1 causation.
Methods and source locations
Compound reported in the paper: Resveratrol; Compound label in the paper: resveratrol
Reported resveratrol experiments; stereochemical purity and equivalence to A10785 are not established.
Materials and methods: Tissue collection, Tissue explants, Western blotting, Glucose uptake, Statistical analysis; Results: skeletal muscle insulin signalling; Fig. 7A–D
Insulin-signalling proteins and glucose uptake: n = 6 patients, mean ± SEM; authors describe restoration to basal levels. No exact fold changes inferred from bars. — Materials and methods: Tissue collection, Tissue explants, Western blotting, Glucose uptake, Statistical analysis; Results: skeletal muscle insulin signalling; Fig. 7A–D
