Journal: Drug Design, Development and Therapy

Article Title: Semaglutide Alleviates Ovary Inflammation via the AMPK/SIRT1/NF‑κB Signaling Pathway in Polycystic Ovary Syndrome Mice

doi: 10.2147/dddt.s484531

Figure Lengend Snippet: Figure 1 (A) Experimental flow chart; (B–D) Three weeks after modeling, PCOS mice were treated with semaglutide for 4 weeks. (B) Body weight; (C) Fasting glucose; (D) IPGTT and AUC. Data are presented as mean ± SD. Vs NC, ##P <0.01; vs PCOS, **P <0.01; n = 6 per group.

Article Snippet: After blocking with skimmed milk at room temperature for 2 hours the membranes were incubated overnight at 4°C with primary antibodies against AMPK (dilution 1:1000; No. 5831S, CST, USA), pAMPK (dilution 1:1000; No. 2535S, CST, USA), SIRT1 (dilution 1:1000; No. 8469S, CST, USA), NF-κB (dilution 1:1000; No. BA0610, BOSTER, China), IκBα (dilution 1:1000; No. 4812S, CST, USA), pIκBα (dilution 1:1000; No. 2859S, CST, USA), IL-1β (dilution 1:1000; No. 516288, ZEN-BIOSCIENCE, China), and IL-6 (dilution 1:1000; No. BA4339, BOSTER, China), TNF-α (dilution 1:1000; No. BA0103, BOSTER, China).

Techniques:

Journal: Drug Design, Development and Therapy

Article Title: Semaglutide Alleviates Ovary Inflammation via the AMPK/SIRT1/NF‑κB Signaling Pathway in Polycystic Ovary Syndrome Mice

doi: 10.2147/dddt.s484531

Figure Lengend Snippet: Figure 4 (A) Immunohistochemical images of IL-1β, IL-6, TNF-α, and NF-κB; Mean density: (B) IL-1β; (C) IL-6; (D) TNF-α; (E) NF-κB; (F) Western blot of IL-1β, IL-6, and TNF-α expression; (G) Peripheral blood IL-1β content. Data are presented as mean ± SD. Vs NC, ##P <0.01; Vs PCOS, *P <0.05, **P <0.01; n = 6 per group.

Article Snippet: After blocking with skimmed milk at room temperature for 2 hours the membranes were incubated overnight at 4°C with primary antibodies against AMPK (dilution 1:1000; No. 5831S, CST, USA), pAMPK (dilution 1:1000; No. 2535S, CST, USA), SIRT1 (dilution 1:1000; No. 8469S, CST, USA), NF-κB (dilution 1:1000; No. BA0610, BOSTER, China), IκBα (dilution 1:1000; No. 4812S, CST, USA), pIκBα (dilution 1:1000; No. 2859S, CST, USA), IL-1β (dilution 1:1000; No. 516288, ZEN-BIOSCIENCE, China), and IL-6 (dilution 1:1000; No. BA4339, BOSTER, China), TNF-α (dilution 1:1000; No. BA0103, BOSTER, China).

Techniques: Immunohistochemical staining, Western Blot, Expressing

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 1. PUN alleviated liver injury in diet-induced NASH. (A) Serum ALT and AST levels and liver hydroxyproline concentrations. Data are presented as the mean ± SD (n = 10, from three independent experiments). (B, C) The H&E staining of liver tissues in the Control, vehicle and PUN (100, 300, 500 mg/kg) groups (original magnification, 200×). (D-E) Detection of ALT and AST concentrations in serum of PUN- treated mice after NASH. (F, G) H&E and Masson staining of liver tissues (original magnification, 200×). Similar results were obtained from three independent experiments. All data are presented as the mean ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs. Control group; ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Staining, Control

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 2. PUN attenuated hepatic inflammatory response in mice with NASH. (A-C) Effects of PUN on NASH- induced serum TNF-α, IL-6, and IL-1β generation. Similar results were obtained from three independent experiments. (D-G) Effects of PUN on NLRP3, ASC, cleaved-caspase-1, and mature-IL-1β protein expression was measured by western blotting and quantification of protein expression was performed by densitometric analysis. Similar results were obtained from three independent experiments. All data are presented as the mean ± SEM (n = 6 in each group). *p < 0.05 and **P < 0.01 vs. Control group; ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Expressing, Western Blot, Control

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 4. PUN upregulated the p62/Nrf-2 signaling in mice with NASH-induced liver injury. (A-C) Immunofluorescence stained analysis of Nrf2 and p-p62 in liver tissues (original magnification, 200×). (D-G) Liver tissue was analyzed by western blot for assessment of nuclear abundance of Nrf2, HO-1 and phosphorylation of p62 at ser349. β-actin was used as an internal control. Similar results were obtained from three independent experiments. All data are presented as means ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs. Control group; #P < 0.05 and ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Immunofluorescence, Staining, Western Blot, Phospho-proteomics, Control

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 5. PUN regulates up-regulated autophagy via the AMPK/mTOR/ULK1 signaling pathway. (A-E) Effects of PUN on protein abundance of Beclin-1, LC3, Atg5 and Atg7 measured by western blotting. (F-I) Effects of PUN on protein abundance of p-AMPK, p-mTOR and p-ULK1 measured by western blotting. Similar results were obtained from three independent experiments. All data are presented as means ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs. Control group; #P < 0.05 and ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Quantitative Proteomics, Western Blot, Control

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 6. PUN improved liver injury caused by NASH dependent upon Nrf2 deficiency in mice. WT and Nrf2−/− mice were injected intraperitoneally with PUN (300 mg/kg) for 24 h followed by challenge with NASH. (A-B) Blood serum collected assessment of ALT and AST activities. (C-F) Liver tissues serum TNF-α, IL-6, and IL-1β and ROS generation. Similar results were obtained from three independent experiments. (G, H) Representative histological sections of liver were stained with H&E and Masson (magnification×400). Similar results were obtained from three independent experiments. All data are presented as means ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs. Control group; #P < 0.05 and ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Injection, Staining, Control

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 7. PUN improved Txnip-NLRP3 signaling pathway caused by NASH dependent upon Nrf2 deficiency in mice. WT and Nrf2−/− mice were injected intraperitoneally with PUN (300 mg/kg) for 24 h followed by challenge with NASH. (A-F) Effects of PUN on protein abundance of Txinp, NLRP3, ASC, Cleaved-Caspase1 and Mature-IL-1β measured by western blot. β-actin was used as an internal control. Similar results were obtained from three independent experiments. All data are presented as means ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs. Control group; #P < 0.05 and ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Injection, Quantitative Proteomics, Western Blot, Control

Journal: Scientific reports

Article Title: Punicalagin relieves hepatic injury by antioxidation and enhancement of autophagy in diet-induced nonalcoholic steatohepatitis.

doi: 10.1038/s41598-025-98044-6

Figure Lengend Snippet: Fig. 8. PUN improved Autophagy caused by NASH dependent upon Nrf2 deficiency in mice. WT and Nrf2−/− mice were injected intraperitoneally with PUN (300 mg/kg) for 24 h followed by challenge with NASH. (A-E) Effects of PUN on protein abundance of Beclin-1, Atg5, Atg7, LC3, p-AMPK, p-ULK1 and p-mTOR measured by western blot. β-actin was used as an internal control. Similar results were obtained from three independent experiments. All data are presented as means ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs. Control group; #P < 0.05 and ##P < 0.01 vs. NASH group.

Article Snippet: Antibodies against Nrf2, SQSTM1/p62 (phospho S349), p-ULK1, p-mTOR and p-AMPK were obtained from Abcam (Cambridge, UK); HO-1, Beclin1, LC3, Atg5, Atg7, NLRP3, ASC, Cleaved Caspase1, IL-1β, ULK1, mTOR and AMPK and β-actin were obtained from Proteintech; Test kits of malondialdehyde (MDA), reactive oxygen species (ROS), GSH, and superoxide dismutase (SOD), alanine transaminase (ALT), aspartate aminotransferase (AST) were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Injection, Quantitative Proteomics, Western Blot, Control