collagen type 1 α1  (Boster Bio)

Journal: Molecular Medicine Reports

Article Title: Acetyl zingerone ameliorates osteoarthritis by inhibiting chondrocyte programmed cell death

doi: 10.3892/mmr.2023.13089

Figure Lengend Snippet: Micro-CT evaluations of AZ-treated OA induced by DMM surgery. (A) Chemical structure of AZ. (B) Nuclear magnetic resonance spectrum of AZ. (C) Micro-CT images of frontal views of the knee joints at 4 weeks after sham or DMM operation. (D) Sagittal views of the medial compartment subchondral bone. Quantitative analysis of (E) BV, (F) BV/TV, (G) Tb. N and (H) Tb. Th. # P<0.05 vs. control; *P<0.05 vs. DMM. AZ, acetyl zingerone; BV, bone volume; DMM, destabilization of the medial meniscus; Tb. N; trabecular number; Tb. Th, trabecular thickness; TV, total volume.

Article Snippet: The following primary and secondary antibodies were used in the present study: Nrf2 (cat. no. 16396-1-AP; 1:2,000), HO-1 (cat. no. 10701-1-AP; 1:1,000), GPX4 (cat. no. 67763-1-lg; 1:2,000), ADAMTS-4 (cat. no. 11865-1-AP; 1:500), inducible nitric oxide synthase (iNOS; cat. no. 18985-1-AP; 1:1,000), MMP13 (cat. no. 18165-1-AP; 1:1,000), cyclooxygenase 2 (COX2; cat. no. 27308-1-AP; 1:1,000), aggrecan (cat. no. 13880-1-AP; 1:1,000), collagen type II α1 (COL2A1; cat. no. 28459-1-AP; 1:1,000), Notch1 (cat. no. 20687-1-AP; 1:1,000), suppressor of cytokine signaling 3 (SOCS3; cat. no. 14025-1-AP; 1:1,000) and β-actin (cat. no. 66009-1-lg; 1:4,000) (all from Proteintech Group Inc.), and caspase 3 (cat. no. A0214; 1:1,000) and cleaved caspase 3 (cat. no. A19654; 1:1,000) (both from ABclonal Biotech Co., Ltd.) primary antibodies; HRP-conjugated AffiniPure goat anti-rabbit IgG (H+L) (cat. no. SA00001-2; 1:8,000) and HRP-conjugated AffiniPure goat anti-mouse IgG (H+L) (cat. no. SA00001-1; 1:8,000) secondary antibodies (both from Proteintech Group Inc.).

Techniques: Micro-CT, Nuclear Magnetic Resonance, Control

Journal: Molecular Medicine Reports

Article Title: Acetyl zingerone ameliorates osteoarthritis by inhibiting chondrocyte programmed cell death

doi: 10.3892/mmr.2023.13089

Figure Lengend Snippet: AZ promotes chondrocyte proliferation, and inhibits the expression of inflammatory factors and matrix-degrading enzymes. Chondrocytes were pretreated with DMSO and IL-1β (20 ng/ml) for 24 h, and were then treated with AZ (25, 50 and 100 µM) for another 24 or 48 h. (A) Cell viability was assessed using the Cell Counting Kit-8 assay. (B) EdU staining (red fluorescence) was used to assess cell proliferation. (C) Expression levels of aggrecan and COL2A1. (D) ImageJ was used to analyze the relative protein expression levels of aggrecan, COL2A1, ADAMTS-4, iNOS, MMP13 and COX2. (E) ADAMTS-4, iNOS, MMP13 and COX2 were detected by western blotting. # P<0.05 vs. control; *P<0.05 vs. IL-1β. AZ, acetyl zingerone; COL2A1, collagen type II α1; COX2, cyclooxygenase 2; EdU, 5-ethynyl-2′-deoxyuridine; iNOS, inducible nitric oxide synthase.

Article Snippet: The following primary and secondary antibodies were used in the present study: Nrf2 (cat. no. 16396-1-AP; 1:2,000), HO-1 (cat. no. 10701-1-AP; 1:1,000), GPX4 (cat. no. 67763-1-lg; 1:2,000), ADAMTS-4 (cat. no. 11865-1-AP; 1:500), inducible nitric oxide synthase (iNOS; cat. no. 18985-1-AP; 1:1,000), MMP13 (cat. no. 18165-1-AP; 1:1,000), cyclooxygenase 2 (COX2; cat. no. 27308-1-AP; 1:1,000), aggrecan (cat. no. 13880-1-AP; 1:1,000), collagen type II α1 (COL2A1; cat. no. 28459-1-AP; 1:1,000), Notch1 (cat. no. 20687-1-AP; 1:1,000), suppressor of cytokine signaling 3 (SOCS3; cat. no. 14025-1-AP; 1:1,000) and β-actin (cat. no. 66009-1-lg; 1:4,000) (all from Proteintech Group Inc.), and caspase 3 (cat. no. A0214; 1:1,000) and cleaved caspase 3 (cat. no. A19654; 1:1,000) (both from ABclonal Biotech Co., Ltd.) primary antibodies; HRP-conjugated AffiniPure goat anti-rabbit IgG (H+L) (cat. no. SA00001-2; 1:8,000) and HRP-conjugated AffiniPure goat anti-mouse IgG (H+L) (cat. no. SA00001-1; 1:8,000) secondary antibodies (both from Proteintech Group Inc.).

Techniques: Expressing, Cell Counting, Staining, Fluorescence, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Acetyl zingerone ameliorates osteoarthritis by inhibiting chondrocyte programmed cell death

doi: 10.3892/mmr.2023.13089

Figure Lengend Snippet: AZ inhibits IL-1β-induced ferroptosis in chondrocytes. (A) GSH and (B) MDA levels were measured in each group using GSH and MDA assays. Chondrocytes were pretreated with IL-1β (20 ng/ml) or Fer-1 (60 nmol/ml) for 24 h and then treated with or without AZ (100 µM) for an additional 24 h. (C) Mitochondria were observed by transmission electron microscopy. (D) GPX4 staining to indicate ferroptosis in mice following DMM surgery. (E) Cells were observed by fluorescence microscopy. (F) Chondrocytes were pretreated with or without IL-1β (20 ng/ml) or Fer-1 (60 nmol/ml) for 24 h and then treated with AZ (100 µM) for another 24 h, and GPX4 expression was determined by western blotting. # P<0.05 vs. control; *P<0.05 vs. IL-1β. AZ, acetyl zingerone; DMM, destabilization of the medial meniscus; Fer-1, ferrostatin-1; GPX4, glutathione peroxidase 4; GSH, glutathione; MDA, malondialdehyde.

Article Snippet: The following primary and secondary antibodies were used in the present study: Nrf2 (cat. no. 16396-1-AP; 1:2,000), HO-1 (cat. no. 10701-1-AP; 1:1,000), GPX4 (cat. no. 67763-1-lg; 1:2,000), ADAMTS-4 (cat. no. 11865-1-AP; 1:500), inducible nitric oxide synthase (iNOS; cat. no. 18985-1-AP; 1:1,000), MMP13 (cat. no. 18165-1-AP; 1:1,000), cyclooxygenase 2 (COX2; cat. no. 27308-1-AP; 1:1,000), aggrecan (cat. no. 13880-1-AP; 1:1,000), collagen type II α1 (COL2A1; cat. no. 28459-1-AP; 1:1,000), Notch1 (cat. no. 20687-1-AP; 1:1,000), suppressor of cytokine signaling 3 (SOCS3; cat. no. 14025-1-AP; 1:1,000) and β-actin (cat. no. 66009-1-lg; 1:4,000) (all from Proteintech Group Inc.), and caspase 3 (cat. no. A0214; 1:1,000) and cleaved caspase 3 (cat. no. A19654; 1:1,000) (both from ABclonal Biotech Co., Ltd.) primary antibodies; HRP-conjugated AffiniPure goat anti-rabbit IgG (H+L) (cat. no. SA00001-2; 1:8,000) and HRP-conjugated AffiniPure goat anti-mouse IgG (H+L) (cat. no. SA00001-1; 1:8,000) secondary antibodies (both from Proteintech Group Inc.).

Techniques: Transmission Assay, Electron Microscopy, Staining, Fluorescence, Microscopy, Expressing, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Acetyl zingerone ameliorates osteoarthritis by inhibiting chondrocyte programmed cell death

doi: 10.3892/mmr.2023.13089

Figure Lengend Snippet: AZ inhibits ferroptosis in osteoarthritis through the Nrf2/HO-1 pathway. Chondrocytes were pretreated with IL-1β (20 ng/ml) for 24 h, with or without ML385 (Nrf2 inhibitors) or Fer-1 (60 nmol/ml) for 12 h, and then treated with or without AZ (100 µM) for an additional 24 h. (A) Protein expression levels of Nrf2, HO-1 and GPX4 was determined by western blotting. (B) Nrf2 staining of ferroptosis in chondrocytes. Chondrocytes were pretreated with IL-1β (20 ng/ml) for 24 h, with or without tin protoporphyrin IX (HO-1 inhibitors) or Fer-1 (60 nmol/ml) for 12 h, and then treated with or without AZ (100 µM) for an additional 24 h. (C) Protein expression levels of Nrf2, HO-1 and GPX4 was determined by western blotting. (D) Schematic diagram showing how AZ affects ferroptosis through the Nrf2/HO-1 signaling pathway. AZ, acetyl zingerone; DMM, destabilization of the medial meniscus; Fer-1, ferrostatin-1; GPX, glutathione peroxidase 4; HO-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2-related factor 2.

Article Snippet: The following primary and secondary antibodies were used in the present study: Nrf2 (cat. no. 16396-1-AP; 1:2,000), HO-1 (cat. no. 10701-1-AP; 1:1,000), GPX4 (cat. no. 67763-1-lg; 1:2,000), ADAMTS-4 (cat. no. 11865-1-AP; 1:500), inducible nitric oxide synthase (iNOS; cat. no. 18985-1-AP; 1:1,000), MMP13 (cat. no. 18165-1-AP; 1:1,000), cyclooxygenase 2 (COX2; cat. no. 27308-1-AP; 1:1,000), aggrecan (cat. no. 13880-1-AP; 1:1,000), collagen type II α1 (COL2A1; cat. no. 28459-1-AP; 1:1,000), Notch1 (cat. no. 20687-1-AP; 1:1,000), suppressor of cytokine signaling 3 (SOCS3; cat. no. 14025-1-AP; 1:1,000) and β-actin (cat. no. 66009-1-lg; 1:4,000) (all from Proteintech Group Inc.), and caspase 3 (cat. no. A0214; 1:1,000) and cleaved caspase 3 (cat. no. A19654; 1:1,000) (both from ABclonal Biotech Co., Ltd.) primary antibodies; HRP-conjugated AffiniPure goat anti-rabbit IgG (H+L) (cat. no. SA00001-2; 1:8,000) and HRP-conjugated AffiniPure goat anti-mouse IgG (H+L) (cat. no. SA00001-1; 1:8,000) secondary antibodies (both from Proteintech Group Inc.).

Techniques: Expressing, Western Blot, Staining