Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression of DDIT4, mTOR, GLUT4, PI3K, and AKT in MHY1485-treated IR C2C12 cells. (A) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, AKT after the activation of mTOR. (B) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, AKT and protein expression levels of DDIT4, GLUT4 after MHY1485 treatment. (C) Glucose levels in culture medium after MHY1485 treatment. (D) Cell TG levels after MHY1485 treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: C2C12 cells were lysed in RIPA lysis buffer (cat. no. C1053-100; Applygen Technologies, Inc.) to extract total cellular proteins.

Techniques: Expressing, Activation Assay, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT in DDIT4-siRNA-treated IR C2C12 cells. (A) DDIT4-siRNA-1 had the most significant silencing effect. (B) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT after DDIT4-siRNA treatment. (C) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, and AKT and protein expression levels of DDIT4 and GLUT4 after DDIT4-siRNA treatment. (D) Glucose levels in culture medium after DDIT4-siRNA treatment. (E) TG levels of C2C12 cells after DDIT4-siRNA treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; siRNA, small interfering RNA; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: C2C12 cells were lysed in RIPA lysis buffer (cat. no. C1053-100; Applygen Technologies, Inc.) to extract total cellular proteins.

Techniques: Expressing, Small Interfering RNA, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression of DDIT4, mTOR, GLUT4, PI3K, and AKT in MHY1485-treated IR C2C12 cells. (A) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, AKT after the activation of mTOR. (B) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, AKT and protein expression levels of DDIT4, GLUT4 after MHY1485 treatment. (C) Glucose levels in culture medium after MHY1485 treatment. (D) Cell TG levels after MHY1485 treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: The C2C12 mouse myoblast cell line (cat. no. CL-0044; Procell Life Science & Technology Co., Ltd.) was cultured in culture medium (cat. no. CM-0044 Procell Life Science & Technology Co., Ltd.) until the cells reached 70-80% confluence.

Techniques: Expressing, Activation Assay, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT in DDIT4-siRNA-treated IR C2C12 cells. (A) DDIT4-siRNA-1 had the most significant silencing effect. (B) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT after DDIT4-siRNA treatment. (C) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, and AKT and protein expression levels of DDIT4 and GLUT4 after DDIT4-siRNA treatment. (D) Glucose levels in culture medium after DDIT4-siRNA treatment. (E) TG levels of C2C12 cells after DDIT4-siRNA treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; siRNA, small interfering RNA; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: The C2C12 mouse myoblast cell line (cat. no. CL-0044; Procell Life Science & Technology Co., Ltd.) was cultured in culture medium (cat. no. CM-0044 Procell Life Science & Technology Co., Ltd.) until the cells reached 70-80% confluence.

Techniques: Expressing, Small Interfering RNA, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression of DDIT4, mTOR, GLUT4, PI3K, and AKT in MHY1485-treated IR C2C12 cells. (A) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, AKT after the activation of mTOR. (B) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, AKT and protein expression levels of DDIT4, GLUT4 after MHY1485 treatment. (C) Glucose levels in culture medium after MHY1485 treatment. (D) Cell TG levels after MHY1485 treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: Differentiated C2C12 cells were then incubated with 0.5 mM palmitic acid (PA; cat. no. 800508; Merck KGaA) for 24 h, designated the PA group.

Techniques: Expressing, Activation Assay, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT in DDIT4-siRNA-treated IR C2C12 cells. (A) DDIT4-siRNA-1 had the most significant silencing effect. (B) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT after DDIT4-siRNA treatment. (C) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, and AKT and protein expression levels of DDIT4 and GLUT4 after DDIT4-siRNA treatment. (D) Glucose levels in culture medium after DDIT4-siRNA treatment. (E) TG levels of C2C12 cells after DDIT4-siRNA treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; siRNA, small interfering RNA; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: Differentiated C2C12 cells were then incubated with 0.5 mM palmitic acid (PA; cat. no. 800508; Merck KGaA) for 24 h, designated the PA group.

Techniques: Expressing, Small Interfering RNA, Control

Journal: Cell Insight

Article Title: Intratumor HIF-1α modulates production of a cachectic ligand to cause host wasting

doi: 10.1016/j.cellin.2025.100247

Figure Lengend Snippet: Tumor-derived IGFBP-5 causes organ wasting. ( A ) Immunoblots indicating IGFBP-5 expression in 3T3-L1 adipocytes, C2C12 myotubes, and C26 cells. ( B ) Differentiation (MHC, red) of C2C12 myoblasts treated with 1 μg/mL IGFBP-5 (BP5) in the differentiation medium for 5 days. ( C ) - ( E ) The widths and morphologies ( C ), gene expression ( D , n = 3), and protein degradation rates indicated by 3 H-Tyrosine release ( E , n = 3) of C2C12 myotubes that were differentiated for 5 days and subsequently treated with fresh differentiation medium containing 1 μg/mL IGFBP-5 (BP5), C26-conditioned differentiation medium (CM), or CM containing 100 ng/mL anti-IGFBP-5 antibodies (Ab) for 2 days. ( F ) Lipolysis rates indicated by released glycerol in differentiated 3T3-L1 adipocytes that were treated 2 μg/mL IGFBP-5 (BP5), or C26-conditioned medium (CM) with or without anti-IGFBP-5 antibodies (Ab) (right, n = 3) for 24 h. ( G ) - ( I ) IGF signaling responses, as indicated by pAkt, in differentiated 3T3-L1 adipocytes that were treated with 100 ng/mL IGF-1 plus 1 μg/mL IGFBP-5 (BP5) in growth medium for 1 h ( G ), or 1 μg/mL IGFBP-5 (BP5) in growth medium at different time points ( H ), or C26-conditioned growth (CM) medium containing anti-IGFBP-5 antibodies (Ab) for 1 h ( I ). ( J ) - ( K ) Immunoblots indicating IGFBP-5 expression in C26 cells ( K ) with frameshift-associated IGFBP5 knockout (BP5KO) caused by a single-base deletion in the Igfbp5 coding region ( J ). ( L ) - ( N ) Tumor weights ( L ), initial injection and post-dissection body weights ( M ), muscle ( N , left) and fat ( N , right) weights of mice bearing control or BP5KO C26 tumors for 25 days ( n = 6). Data were presented as means ± SEM. Unpaired Student's t -test and one-way ANOVA followed by post hoc test were performed to assess differences. ∗p < 0.05.

Article Snippet: Both mouse pre-adipocyte 3T3-L1 (ATCC) and myoblast C2C12 cells (ATCC) were cultured in growth medium (DMEM, 10% FBS, antibiotics).

Techniques: Derivative Assay, Western Blot, Expressing, Gene Expression, Knock-Out, Injection, Dissection, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Primers used for real-time quantitative polymerase chain reaction.

Article Snippet: Some of the PA + RSV group cells were again divided into two groups: The PA + RSV + MHY1485 group C2C12 cells were treated with 10 µM MHY1485 (cat. no. HY-B0795; MedChemExpress) to activate mTOR, and the PA + RSV + DDIT4-siRNA group C2C12 cells were transfected with a small interfering (si)RNA against DDIT4 (DDIT4-siRNA; Shanghai Genechem Co. Ltd.), all incubated at 37˚C for 24 h. The cells were collected for western blotting and cellular TG content determination.

Techniques: Sequencing

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Effects of RSV on the mRNA and protein expression levels of DDIT4 in skeletal muscle. (A) Relative mRNA expression of DDIT4. (B) Protein bands of DDIT4. (C) DDIT4/β-actin protein expression levels. Data are presented as the mean ± SD (n=11). * P<0.05 vs. the CON group; # P<0.05 vs. the HFD group. RSV, resveratrol; DDIT4, DNA-damage-inducible transcript 4; CON, control; HFD, high-fat diet.

Article Snippet: Some of the PA + RSV group cells were again divided into two groups: The PA + RSV + MHY1485 group C2C12 cells were treated with 10 µM MHY1485 (cat. no. HY-B0795; MedChemExpress) to activate mTOR, and the PA + RSV + DDIT4-siRNA group C2C12 cells were transfected with a small interfering (si)RNA against DDIT4 (DDIT4-siRNA; Shanghai Genechem Co. Ltd.), all incubated at 37˚C for 24 h. The cells were collected for western blotting and cellular TG content determination.

Techniques: Expressing, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression of DDIT4, mTOR, GLUT4, PI3K, and AKT in MHY1485-treated IR C2C12 cells. (A) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, AKT after the activation of mTOR. (B) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, AKT and protein expression levels of DDIT4, GLUT4 after MHY1485 treatment. (C) Glucose levels in culture medium after MHY1485 treatment. (D) Cell TG levels after MHY1485 treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: Some of the PA + RSV group cells were again divided into two groups: The PA + RSV + MHY1485 group C2C12 cells were treated with 10 µM MHY1485 (cat. no. HY-B0795; MedChemExpress) to activate mTOR, and the PA + RSV + DDIT4-siRNA group C2C12 cells were transfected with a small interfering (si)RNA against DDIT4 (DDIT4-siRNA; Shanghai Genechem Co. Ltd.), all incubated at 37˚C for 24 h. The cells were collected for western blotting and cellular TG content determination.

Techniques: Expressing, Activation Assay, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT in DDIT4-siRNA-treated IR C2C12 cells. (A) DDIT4-siRNA-1 had the most significant silencing effect. (B) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT after DDIT4-siRNA treatment. (C) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, and AKT and protein expression levels of DDIT4 and GLUT4 after DDIT4-siRNA treatment. (D) Glucose levels in culture medium after DDIT4-siRNA treatment. (E) TG levels of C2C12 cells after DDIT4-siRNA treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; siRNA, small interfering RNA; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: Some of the PA + RSV group cells were again divided into two groups: The PA + RSV + MHY1485 group C2C12 cells were treated with 10 µM MHY1485 (cat. no. HY-B0795; MedChemExpress) to activate mTOR, and the PA + RSV + DDIT4-siRNA group C2C12 cells were transfected with a small interfering (si)RNA against DDIT4 (DDIT4-siRNA; Shanghai Genechem Co. Ltd.), all incubated at 37˚C for 24 h. The cells were collected for western blotting and cellular TG content determination.

Techniques: Expressing, Small Interfering RNA, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Resveratrol ameliorates IR via the DDIT4/mTOR/PI3K/AKT/GLUT4 pathway. IR, insulin resistance; DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; GLUT4, glucose transporter 4; PA, palmitic acid.

Article Snippet: Some of the PA + RSV group cells were again divided into two groups: The PA + RSV + MHY1485 group C2C12 cells were treated with 10 µM MHY1485 (cat. no. HY-B0795; MedChemExpress) to activate mTOR, and the PA + RSV + DDIT4-siRNA group C2C12 cells were transfected with a small interfering (si)RNA against DDIT4 (DDIT4-siRNA; Shanghai Genechem Co. Ltd.), all incubated at 37˚C for 24 h. The cells were collected for western blotting and cellular TG content determination.

Techniques: