Journal: Experimental and Therapeutic Medicine

Article Title: Resveratrol‑mediated activation of SIRT1 inhibits the PERK‑eIF2α‑ATF4 pathway and mitigates bupivacaine‑induced neurotoxicity in PC12 cells

doi: 10.3892/etm.2023.12132

Figure Lengend Snippet: RSV protects PC12 cells against BUP-induced cytotoxicity via SIRT1 upregulation. (A) Cell viability in PC12 cells exposed to increasing concentrations of EX527. (B) Morphology of PC12 cells in the control, BUP, RSV + BUP and EX527 + RSV + BUP groups observed under a phase-contrast microscope (magnification, x200; scale bar, 50 µm). (C) Representative western blot images and semi-quantification of SIRT1 protein levels in each group. (D) Cell viability in each group. (E) Representative immunofluorescence images of SIRT1 (green) and cell nuclei (blue) staining (scale bar, 50 µm) and the relative mean intensity of SIRT1 immunofluorescence in each group. Data are presented as the mean ± SEM (n=3). * P<0.05 vs. the control group; # P<0.05 vs. the BUP group; & P<0.05 vs. the RSV + BUP group. RSV, resveratrol; BUP, bupivacaine, SIRT1, sirtuin 1; EX527, SIRT1 inhibitor.

Article Snippet: BUP hydrochloride and RSV were purchased from Sigma-Aldrich (Merck KGaA), while CCT020312 and EX527 were obtained from MedChemExpress.

Techniques: Control, Microscopy, Western Blot, Immunofluorescence, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Resveratrol‑mediated activation of SIRT1 inhibits the PERK‑eIF2α‑ATF4 pathway and mitigates bupivacaine‑induced neurotoxicity in PC12 cells

doi: 10.3892/etm.2023.12132

Figure Lengend Snippet: RSV protects against BUP-induced apoptosis via the upregulation of SIRT1 expression. (A) Representative western blot images and semi-quantification of Bax, Bcl-2 and cleaved caspase-3 protein levels in the control, BUP, RSV + BUP and EX527 + RSV + BUP groups. Q1 (upper-left quadrant) represents dead cells; Q2 (upper-right quadrant) represents late apoptotic cells; Q3 (lower-right quadrant) represents early apoptotic cells; Q4 (lower-left quadrant) represents live cells. (B) Representative images of the flow cytometric analysis of Annexin V/7-AAD staining and quantification of the apoptotic rates in each group. Data are presented as the mean ± SEM (n=3). * P<0.05 vs. the control group; # P<0.05 vs. the BUP group; & P<0.05 vs. the RSV + BUP group. RSV, resveratrol; BUP, bupivacaine; SIRT1, sirtuin 1; EX527, SIRT1 inhibitor; 7-AAD, 7-aminoactinomycin D.

Article Snippet: BUP hydrochloride and RSV were purchased from Sigma-Aldrich (Merck KGaA), while CCT020312 and EX527 were obtained from MedChemExpress.

Techniques: Expressing, Western Blot, Control, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Resveratrol‑mediated activation of SIRT1 inhibits the PERK‑eIF2α‑ATF4 pathway and mitigates bupivacaine‑induced neurotoxicity in PC12 cells

doi: 10.3892/etm.2023.12132

Figure Lengend Snippet: RSV inhibits the PERK-eIF2α-ATF4 pathway via the upregulation of SIRT1 expression in BUP-treated PC12 cells. (A) Representative western blot images and semi-quantification of p-PERK/PERK, p-eIF2α/eIF2α and ATF4 protein levels in the control, BUP, RSV + BUP and EX527 + RSV + BUP groups. (B) Representative immunofluorescence images of ATF4 (red) and cell nuclei (blue) staining (scale bar, 50 µm) and relative mean intensity analysis of ATF4 immunofluorescence in each group. Data are presented as the mean ± SEM (n=3). * P<0.05 vs. the control group; # P<0.05 vs. the BUP group; & P<0.05 vs. the RSV+BUP group. RSV, resveratrol; PERK, protein kinase RNA-like ER kinase; eIF2α, eukaryotic translation initiation factor 2 α; ATF4, activating transcription factor 4; SIRT1, sirtuin 1; BUP, bupivacaine; p-, phosphorylated; EX527, SIRT1 inhibitor.

Article Snippet: BUP hydrochloride and RSV were purchased from Sigma-Aldrich (Merck KGaA), while CCT020312 and EX527 were obtained from MedChemExpress.

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Staining

Journal: PLoS ONE

Article Title: Synergistic Interactions between HDAC and Sirtuin Inhibitors in Human Leukemia Cells

doi: 10.1371/journal.pone.0022739

Figure Lengend Snippet: Primary AML cells were incubated with or without sirtuin inhibitors (EX527, sirtinol, cambinol) in the presence or absence of VA at the indicated concentrations. Viability was assessed 48 h later by PI cell staining and flow cytometry. CI values refer to the highest drug concentrations used. CI CT s for each drug combination are presented in the lower insets. An effect of 1 corresponds to 100% specific death whereas an effect of 0 corresponds to 0% specific death.

Article Snippet: EX527 was from Tocris Bioscience (Bristol, UK).

Techniques: Incubation, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Synergistic Interactions between HDAC and Sirtuin Inhibitors in Human Leukemia Cells

doi: 10.1371/journal.pone.0022739

Figure Lengend Snippet: A, 3×10 6 primary AML cells/well were plated in 6-well plates and incubated in the presence or absence of 50 µM cambinol, 100 µg/ml VA, or their combination. ΔΨ m was monitored at the indicated time points by TMRE staining and flow cytometry. B, 1×10 6 Jurkat cells were plated in 6-well plates and incubated for 48 h in the presence or absence of 100 µg/ml VA. Thereafter, intracellular Bax content was determined by flow cytometry. C, D, Jurkat cells were transduced with either pMIG or pMIG-Bax. Infected cells were FACS sorted, allowed to expand, and subsequently used for flow cytometric detection of intracellular Bax (C) and for viability assays (D). For these, pMIG- or pMIG-Bax-transduced Jurkat were plated in 96-well plates and incubated in the presence or absence of EX527 or cambinol at the indicated concentrations. Viability was determined by PI staining and flow cytometry 48 h later. A–C, one representative experiment out of three is presented. D, Results are means ± SD of three separate experiments.

Article Snippet: EX527 was from Tocris Bioscience (Bristol, UK).

Techniques: Incubation, Staining, Flow Cytometry, Transduction, Infection

Journal: PLoS ONE

Article Title: Synergistic Interactions between HDAC and Sirtuin Inhibitors in Human Leukemia Cells

doi: 10.1371/journal.pone.0022739

Figure Lengend Snippet: A, B, U937 and 697 cells were transduced to either express an anti-EGFP shRNA (EGFP-sh) or a validated anti-Bax shRNA (Bax-sh). Thereafter cells were used for cell lysate preparation and Bax and γ-tubulin were detected by immunoblotting. B, EGFP-sh or Bax-sh 697 cells were incubated with or without 100 µg/ml VA, 75 µM EX527, 50 µM cambinol, or their combinations. Viability was assessed 36 h later by PI staining and flow cytometry. C, D, EGFP-sh or Bax-sh U937 cells were incubated with or without 100 µg/ml VA, 150 µM EX527, 100 µM cambinol, or their combinations. 36 h later, cells were imaged by light microscopy (D) and cell death was determined by flow cytometric quantification of PI-positive cells (C). A, D, one representative experiment out of three is presented. B, C, Results are means ± SD of three separate experiments. *: p<0.05.

Article Snippet: EX527 was from Tocris Bioscience (Bristol, UK).

Techniques: shRNA, Western Blot, Incubation, Staining, Flow Cytometry, Light Microscopy

Journal: Scientific Reports

Article Title: Marein from Coreopsis tinctoria Nutt. alleviates oxidative stress and lipid accumulation via SIRT1/Nrf2 signaling

doi: 10.1038/s41598-025-97964-7

Figure Lengend Snippet: Marein restored SIRT1/Nrf2 signaling in H 2 O 2 -induced HepG2 cells. HepG2 cells were exposed 500μM H 2 O 2 condition for 24h, and then administrated by 5 μM Marein for another 24h. ( a , b ) mRNA expression levels of SIRT1 ( a ) and Nrf2 ( b ) were assessed using RT-qPCR. ( c ) Western blot analysis of SIRT1 and Nrf2 protein levels. ( d ) Immunofluorescence staining of Nrf2 nuclear localization (scale bar = 25 μm). Data are represented as mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Besides, HepG2 cells was incubated with 5 μM Marein (≥ 96% purity, Shanghai Yuanye Bio-Technology Co., Ltd., China) for 24 h . For inhibitor experiments, cells were pre-treated with SIRT1 inhibitor EX-527 (100 nM, #A10377, AdooQ BioScience, Nanjing, China) for 2 h before Marein treatment.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Marein from Coreopsis tinctoria Nutt. alleviates oxidative stress and lipid accumulation via SIRT1/Nrf2 signaling

doi: 10.1038/s41598-025-97964-7

Figure Lengend Snippet: Inactivation of SIRT1/Nrf2 reversed the antioxidant ability of marein in H 2 O 2 -induced HepG2 cells. HepG2 cells were subjected to different groups: control, H 2 O 2 , H 2 O 2 + Marein, H 2 O 2 + Marein + EX-527 (SIRT1 inhibitor). ( a ) CCK-8 detection was used to examine cell viability. ( b ) LDH content was detected to examine oxidative stress injury. ( c – e ) MDA ( c ), SOD ( d ), and GSH-Px ( e ) levels were detected by ELISA. ( f ) DCFDA detected ROS level. Data are represented as mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Besides, HepG2 cells was incubated with 5 μM Marein (≥ 96% purity, Shanghai Yuanye Bio-Technology Co., Ltd., China) for 24 h . For inhibitor experiments, cells were pre-treated with SIRT1 inhibitor EX-527 (100 nM, #A10377, AdooQ BioScience, Nanjing, China) for 2 h before Marein treatment.

Techniques: Control, CCK-8 Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Marein from Coreopsis tinctoria Nutt. alleviates oxidative stress and lipid accumulation via SIRT1/Nrf2 signaling

doi: 10.1038/s41598-025-97964-7

Figure Lengend Snippet: Inactivation of SIRT1/Nrf2 restrained protective role of marein in H 2 O 2 -triggered lipid accumulation. HepG2 cells were subjected to different groups: control, H 2 O 2 , H 2 O 2 + Marein, H 2 O 2 + Marein + EX-527 (SIRT1 inhibitor). ( a – d ) Determination of TC ( a ), TG ( b ), LDL-C ( c ), and HDL-C ( d ) levels by ELISA. ( e , f ) Detection of lipid metabolism related genes HMGCR ( e ) and LDLR ( f ) by RT-qPCR. ( g ) Western blot analysis of HMGCR and LDLR protein levels. Values were expressed as mean ± SD of three separate determinations. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Besides, HepG2 cells was incubated with 5 μM Marein (≥ 96% purity, Shanghai Yuanye Bio-Technology Co., Ltd., China) for 24 h . For inhibitor experiments, cells were pre-treated with SIRT1 inhibitor EX-527 (100 nM, #A10377, AdooQ BioScience, Nanjing, China) for 2 h before Marein treatment.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: COX-2/sEH Dual Inhibitor Alleviates Hepatocyte Senescence in NAFLD Mice by Restoring Autophagy through Sirt1/PI3K/AKT/mTOR

doi: 10.3390/ijms23158267

Figure Lengend Snippet: PTUPB enhances hepatocyte autophagy by promoting Sirt1 expression. The protein expressions of Sirt1 in the liver (( A , B ), n = 6) and AML12 cells (( C , D ), n = 3) were measured by Western blot and quantitatively analyzed. Cells were treated with PTUPB (1 μM) or EX527 (10 μM) for 1 h before treatment with PA (200 µM) for 48 h. The protein expressions of Sirt1, γH2AX, Fundc1, and LC3II/I in AML12 cells were measured by Western blot and quantitatively analyzed (( E – I ), n = 3). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: An autophagy inhibitor 3-MA (5 mM, MCE, Dallas, TX, USA) or Sirt1 inhibitor EX527 (10 μM, Topscience, Shandong, China) was used to clarify the role of autophagy or Sirt1 in the anti-senescence effects of PTUPB in AML12 cells.

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: COX-2/sEH Dual Inhibitor Alleviates Hepatocyte Senescence in NAFLD Mice by Restoring Autophagy through Sirt1/PI3K/AKT/mTOR

doi: 10.3390/ijms23158267

Figure Lengend Snippet: PTUPB inhibits the PI3K/AKT/mTOR signaling pathway by promoting Sirt1 expression. The mTOR protein and its phosphorylation level in the liver were measured by Western blot and quantitatively analyzed (( A – C ), n = 6). Cells were treated with PTUPB (1 μM) for 1 h before treatment with PA (200 µM). The levels of p-mTOR, p-AKT, and p-PI3K in AML12 cells were measured by Western blot and quantitatively analyzed after PA stimulation for 30 min (( D – G ), n = 3). Cells were treated with PTUPB (1 μM) or EX527 (10 μM) for 1 h before treatment with PA (200 µM). The levels of p-mTOR, p-AKT, and p-PI3K in AML12 cells were measured by Western blot and quantitatively analyzed after PA stimulation for 30 min (( H – K ), n = 3). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: An autophagy inhibitor 3-MA (5 mM, MCE, Dallas, TX, USA) or Sirt1 inhibitor EX527 (10 μM, Topscience, Shandong, China) was used to clarify the role of autophagy or Sirt1 in the anti-senescence effects of PTUPB in AML12 cells.

Techniques: Expressing, Phospho-proteomics, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: COX-2/sEH Dual Inhibitor Alleviates Hepatocyte Senescence in NAFLD Mice by Restoring Autophagy through Sirt1/PI3K/AKT/mTOR

doi: 10.3390/ijms23158267

Figure Lengend Snippet: Antibodies were used in this study.

Article Snippet: An autophagy inhibitor 3-MA (5 mM, MCE, Dallas, TX, USA) or Sirt1 inhibitor EX527 (10 μM, Topscience, Shandong, China) was used to clarify the role of autophagy or Sirt1 in the anti-senescence effects of PTUPB in AML12 cells.

Techniques: