Journal: Lipids in health and disease

Article Title: Puerarin ameliorates metabolic dysfunction-associated fatty liver disease by inhibiting ferroptosis and inflammation.

doi: 10.1186/s12944-023-01969-y

Figure Lengend Snippet: Fig. 3 Puerarin inhibits hepatic ferroptosis and activates the SIRT1/Nrf2 signaling pathway in MAFLD mice. (A) The protein levels of SIRT1, p-Nrf2, Nrf2, HO-1, SLC7A11, and GPX4 were determined by Western blotting. (B) Representative images of immunohistochemical staining showing the expression of SIRT1 in liver sections. (C) Molecular docking model of SIRT1 and Puerarin (i. Spherical diagram, ii. 3D structures of the binding interface, iii. Puerarin was predicted to interact with SIRT1 via the amino acids PRO271, GLN345, ASN346, HIS363, GLN294, LYS444, and VAL445 via hydrophobic interactions or hydrogen bonds. The data are expressed as the mean ± SD (n = 5). *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The specific primary antibodies were as follows: SIRT1 (1:1000, Abcam, USA, Cat. No. ab189494), p-Nrf2 (Ser 40) (1:1000, Affinity, China, Cat. No. DF7519), Nrf2 (1:1000, CST, USA, Cat. No. 12,721), HO-1 (1:1000, ABclonal, China, Cat. No. A1346), SLC7A11 (1:1000, ABclonal, China, Cat. No. A2413), GPX4 (1:1000, Abcam, USA, Cat. No. ab125066), phospho-NF-κB p65 (1:1000, CST, USA, Cat. No. 3033T), NF-κB p65 (1:1000, CST, USA, Cat. No. 8242T), TNF-α (1:1000, Bioss, China, Cat. No. bs-2081R), IL-1β (1:100, Abcam, USA, Cat. No. ab283818), IL-6 (1:100, Abcam, USA, Cat. No. ab290735), and β-actin (1:1000, Abways, China, Cat. No. Ab0035).

Techniques: Western Blot, Immunohistochemical staining, Staining, Expressing, Binding Assay

Journal: Lipids in health and disease

Article Title: Puerarin ameliorates metabolic dysfunction-associated fatty liver disease by inhibiting ferroptosis and inflammation.

doi: 10.1186/s12944-023-01969-y

Figure Lengend Snippet: Fig. 4 Puerarin increases Nrf2 nuclear translocation and reduces liver inflammation in MAFLD mice. (A) Representative images of immunofluorescence staining of p-Nrf2. Scale Bar, 20 μm. (B) F4/80, IL-1β, and IL-6 were determined by immunohistochemical staining. Scale Bar, 50 μm. (C) The protein levels of p-p65/p65, TNF-α, IL-1β, and IL-6 were determined by Western blotting. (D) Hepatic mRNA levels of Tnf-α, Il-1β, and Il-6 were determined by RT‒qPCR. The data are expressed as the mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The specific primary antibodies were as follows: SIRT1 (1:1000, Abcam, USA, Cat. No. ab189494), p-Nrf2 (Ser 40) (1:1000, Affinity, China, Cat. No. DF7519), Nrf2 (1:1000, CST, USA, Cat. No. 12,721), HO-1 (1:1000, ABclonal, China, Cat. No. A1346), SLC7A11 (1:1000, ABclonal, China, Cat. No. A2413), GPX4 (1:1000, Abcam, USA, Cat. No. ab125066), phospho-NF-κB p65 (1:1000, CST, USA, Cat. No. 3033T), NF-κB p65 (1:1000, CST, USA, Cat. No. 8242T), TNF-α (1:1000, Bioss, China, Cat. No. bs-2081R), IL-1β (1:100, Abcam, USA, Cat. No. ab283818), IL-6 (1:100, Abcam, USA, Cat. No. ab290735), and β-actin (1:1000, Abways, China, Cat. No. Ab0035).

Techniques: Translocation Assay, Immunofluorescence, Staining, Immunohistochemical staining, Western Blot

Journal: Lipids in health and disease

Article Title: Puerarin ameliorates metabolic dysfunction-associated fatty liver disease by inhibiting ferroptosis and inflammation.

doi: 10.1186/s12944-023-01969-y

Figure Lengend Snippet: Fig. 5 Puerarin inhibits ferroptosis, activates the SIRT1/Nrf2 signaling pathway, and reduces inflammation in PA-induced AML12 cells in vitro. (A) Co immunoprecipitation was performed to assess the interaction between SIRT1 and Nrf2 in AML12 cells. (B) Protein levels of SIRT1, p-Nrf2, Nrf2, HO-1, SLC7A11, and GPX4 in AML12 cells in the Control + PBS, Control + Puerarin, PA + PBS, and PA + Puerarin groups. (C) Coimmunoprecipitation was performed to assess the interaction between SIRT1 and p65 in AML12 cells. (D) Protein levels of p-p65/p65, TNF-α, IL-1β, and IL-6 in the four groups. (E) Representative images of immunofluorescence staining of p-Nrf2 in AML12 cells. Scale Bar, 20 μm. (F) Representative images of Ferrorange staining in the four groups. Scale Bar, 20 μm. The data are expressed as the mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The specific primary antibodies were as follows: SIRT1 (1:1000, Abcam, USA, Cat. No. ab189494), p-Nrf2 (Ser 40) (1:1000, Affinity, China, Cat. No. DF7519), Nrf2 (1:1000, CST, USA, Cat. No. 12,721), HO-1 (1:1000, ABclonal, China, Cat. No. A1346), SLC7A11 (1:1000, ABclonal, China, Cat. No. A2413), GPX4 (1:1000, Abcam, USA, Cat. No. ab125066), phospho-NF-κB p65 (1:1000, CST, USA, Cat. No. 3033T), NF-κB p65 (1:1000, CST, USA, Cat. No. 8242T), TNF-α (1:1000, Bioss, China, Cat. No. bs-2081R), IL-1β (1:100, Abcam, USA, Cat. No. ab283818), IL-6 (1:100, Abcam, USA, Cat. No. ab290735), and β-actin (1:1000, Abways, China, Cat. No. Ab0035).

Techniques: In Vitro, Immunoprecipitation, Control, Immunofluorescence, Staining

Journal: Lipids in health and disease

Article Title: Puerarin ameliorates metabolic dysfunction-associated fatty liver disease by inhibiting ferroptosis and inflammation.

doi: 10.1186/s12944-023-01969-y

Figure Lengend Snippet: Fig. 6 SIRT1 inhibition reduced the protective effect of puerarin on PA-induced AML12 cells. (A) Protein levels of SIRT1, p-Nrf2, Nrf2, HO-1, SLC7A11, and GPX4 in AML12 cells in the Control + PBS, Control + Puerarin, PA + PBS, PA + Puerarin, PA + EX-527 (10 μM), and PA + Puerarin + EX-527 (10 μM) groups. (B) Representative images of immunofluorescence staining of p-Nrf2 in AML12 cells. Scale Bar, 20 μm. (C) The protein levels of p-p65/p65, TNF-α, IL-1β, and IL-6 in AML12 cells in the six groups. (D) The mRNA levels of Tnf-α, Il-1β, and Il-6 in AML12 cells in the six groups. The data are expressed as the mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The specific primary antibodies were as follows: SIRT1 (1:1000, Abcam, USA, Cat. No. ab189494), p-Nrf2 (Ser 40) (1:1000, Affinity, China, Cat. No. DF7519), Nrf2 (1:1000, CST, USA, Cat. No. 12,721), HO-1 (1:1000, ABclonal, China, Cat. No. A1346), SLC7A11 (1:1000, ABclonal, China, Cat. No. A2413), GPX4 (1:1000, Abcam, USA, Cat. No. ab125066), phospho-NF-κB p65 (1:1000, CST, USA, Cat. No. 3033T), NF-κB p65 (1:1000, CST, USA, Cat. No. 8242T), TNF-α (1:1000, Bioss, China, Cat. No. bs-2081R), IL-1β (1:100, Abcam, USA, Cat. No. ab283818), IL-6 (1:100, Abcam, USA, Cat. No. ab290735), and β-actin (1:1000, Abways, China, Cat. No. Ab0035).

Techniques: Inhibition, Control, Immunofluorescence, Staining

Journal: Lipids in health and disease

Article Title: Puerarin ameliorates metabolic dysfunction-associated fatty liver disease by inhibiting ferroptosis and inflammation.

doi: 10.1186/s12944-023-01969-y

Figure Lengend Snippet: Fig. 7 Nrf2 knockdown reduces puerarin-mediated inhibition of ferroptosis in PA-induced AML12 cells. A. Protein levels of p-Nrf2, Nrf2, HO-1, SLC7A11, and GPX4 in AML12 cells in the siRNA-NC, siRNA-NC + puerarin, siRNA-Nrf2, and siRNA-Nrf2 + puerarin groups. The data are expressed as the mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The specific primary antibodies were as follows: SIRT1 (1:1000, Abcam, USA, Cat. No. ab189494), p-Nrf2 (Ser 40) (1:1000, Affinity, China, Cat. No. DF7519), Nrf2 (1:1000, CST, USA, Cat. No. 12,721), HO-1 (1:1000, ABclonal, China, Cat. No. A1346), SLC7A11 (1:1000, ABclonal, China, Cat. No. A2413), GPX4 (1:1000, Abcam, USA, Cat. No. ab125066), phospho-NF-κB p65 (1:1000, CST, USA, Cat. No. 3033T), NF-κB p65 (1:1000, CST, USA, Cat. No. 8242T), TNF-α (1:1000, Bioss, China, Cat. No. bs-2081R), IL-1β (1:100, Abcam, USA, Cat. No. ab283818), IL-6 (1:100, Abcam, USA, Cat. No. ab290735), and β-actin (1:1000, Abways, China, Cat. No. Ab0035).

Techniques: Knockdown, Inhibition

Journal: Lipids in health and disease

Article Title: Puerarin ameliorates metabolic dysfunction-associated fatty liver disease by inhibiting ferroptosis and inflammation.

doi: 10.1186/s12944-023-01969-y

Figure Lengend Snippet: Fig. 8 Proposed antiferroptotic and anti-inflammatory mechanism of puerarin. Puerarin ameliorates metabolic dysfunction-associated fatty liver disorder by suppressing ferroptosis and inflammation via the SIRT1/Nrf2 signaling pathway

Article Snippet: The specific primary antibodies were as follows: SIRT1 (1:1000, Abcam, USA, Cat. No. ab189494), p-Nrf2 (Ser 40) (1:1000, Affinity, China, Cat. No. DF7519), Nrf2 (1:1000, CST, USA, Cat. No. 12,721), HO-1 (1:1000, ABclonal, China, Cat. No. A1346), SLC7A11 (1:1000, ABclonal, China, Cat. No. A2413), GPX4 (1:1000, Abcam, USA, Cat. No. ab125066), phospho-NF-κB p65 (1:1000, CST, USA, Cat. No. 3033T), NF-κB p65 (1:1000, CST, USA, Cat. No. 8242T), TNF-α (1:1000, Bioss, China, Cat. No. bs-2081R), IL-1β (1:100, Abcam, USA, Cat. No. ab283818), IL-6 (1:100, Abcam, USA, Cat. No. ab290735), and β-actin (1:1000, Abways, China, Cat. No. Ab0035).

Techniques:

Journal: Journal of Ophthalmology

Article Title: Maslinic Acid Protects against Streptozotocin-Induced Diabetic Retinopathy by Activating Nrf2 and Suppressing NF- κ B

doi: 10.1155/2022/3044202

Figure Lengend Snippet: The nuclear levels of Nrf2 as detected by ELISA and by western blotting in the retina of all groups of rats. Data were analyzed by 1-way ANOVA, and all results were presented as mean ± SD ( n = 8). Values are considered significantly different at P < 0.05. a : significantly different as compared to control rats. b : significantly different as compared to MA-treated rats. c : significantly different as compared to T1DM-induced rats.

Article Snippet: The membranes were then washed with the TBST buffer 3 times each of 10 min at room temperature and then incubated with the primary monoclonal antibodies against Bcl2 (Cat sc-7382, 1 : 1000, 26 kDa), Bax (Cat sc-7480, 1 : 1000, 23 kDa, lamin A (sc-518013, 1 : 1000; 69 kDa) (Santa Cruz Biotechnology, USA), cleaved caspase-3 (Cat 9661, 1 : 500, 17/19 kDa), Nrf2 (Cat 12721, 1 : 500, 100 kDa), and β -actin (Cat. No. 47778, 1 : 10000, 45 kDa) (Cell Signaling Technology, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot