Journal: Drug Design, Development and Therapy

Article Title: Tomatidine Alleviates Intervertebral Disc Degeneration by Activating the Nrf2/HO-1/GPX4 Signaling Pathway

doi: 10.2147/DDDT.S481714

Figure Lengend Snippet: The effect of Td on the viability and proliferation of NPCs. ( A ) The chemical structure of Td. ( B ) The 3D structure image of Td from PubChem. ( C ) Cell counting Kit-8 assay results of NPCs stimulated with Td at different concentrations (0,1,2,4,8,16,32,64,128,256) μM at 24h. ( D ) Cell counting Kit-8 assay results of NPCs stimulated with Td at different concentrations (0,1,2,4,8,16,32,64,128,256) uM at 48h. ( E ) The results of the Cell Counting Kit-8 assay for LPS-induced NPCs after 24 hours of Td treatment. ( F ) The results of the Cell Counting Kit-8 assay for LPS-induced NPCs after 48 hours of Td treatment. All data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001, n = 5.

Article Snippet: Tomatidine (Td, purity ≥98%) and lipopolysaccharide (LPS, purity >99%) were purchased from MCE (China).

Techniques: Cell Counting

Journal: Drug Design, Development and Therapy

Article Title: Tomatidine Alleviates Intervertebral Disc Degeneration by Activating the Nrf2/HO-1/GPX4 Signaling Pathway

doi: 10.2147/DDDT.S481714

Figure Lengend Snippet: The effect of Td on ECM synthesis and catabolism of NPCs in vitro.( A ) Toluidine blue staining for the high-density culture of NPCs for 1 week (scale bar: 1mm). ( B ) The integrated optical density (IOD) value of NPCs was analyzed using Image J. ( C ) The gene expression ADAMTS-4, Col-2 and MMP3 were detected by qPCR in the NPCs, treated with or without the administration of LPS for 24h. ( D and E )Western blot analysis of ADAMTS-4, Aggrecan, Col-2, MMP3 and GAPDH expression in NPCs stimulated with LPS (100 ng/mL). The protein expression were evaluated by Image J. All data are presented as mean ± SD. **P < 0.01, ***P < 0.001 and ****P < 0.0001, n = 5.

Article Snippet: Tomatidine (Td, purity ≥98%) and lipopolysaccharide (LPS, purity >99%) were purchased from MCE (China).

Techniques: In Vitro, Staining, Gene Expression, Western Blot, Expressing

Journal: Drug Design, Development and Therapy

Article Title: Tomatidine Alleviates Intervertebral Disc Degeneration by Activating the Nrf2/HO-1/GPX4 Signaling Pathway

doi: 10.2147/DDDT.S481714

Figure Lengend Snippet: Protective effect of Td against oxidative stress and ferroptosis in NPCs. ( A ) The intracellular ROS levels in NPCs were measured by flow cytometry assay. ( B )The levels of GSH and MDA in NPCs were determined by ELISA. ( C ) The content of ferrous iron (Fe2+) within cells was assessed using the Cell Ferrous Iron (Fe2+) Fluorometric Assay Kit. ( D and E ) Western blot analysis of TFR, xCT, SLC40A1, FTL and GAPDH expression in NPCs stimulated with LPS (100 ng/mL). The protein expression were evaluated by Image J. ( F and G ) The protein expression of TFR was evaluated by cell immunofluorescence (scale bar: 10 μm) and Image J. All data are presented as mean ± SD. **P < 0.01, ***P < 0.001 and ****P < 0.0001, n = 5.

Article Snippet: Tomatidine (Td, purity ≥98%) and lipopolysaccharide (LPS, purity >99%) were purchased from MCE (China).

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence

Journal: Drug Design, Development and Therapy

Article Title: Tomatidine Alleviates Intervertebral Disc Degeneration by Activating the Nrf2/HO-1/GPX4 Signaling Pathway

doi: 10.2147/DDDT.S481714

Figure Lengend Snippet: Td can target the Nrf2/HO-1/GPX4 axis for potential effects. ( A )Results of Td force analysis of 2D images. ( B ) The 3D structure image of Td interacting with Nrf2 in the image. ( C ) The gene expression Nrf2 and GPX4 were detected by qPCR in the NPCs, treated with or without the administration of LPS for 24h. ( D and E ) Western blot analysis of Nrf2, HO-1, GPX4 and GAPDH expression in NPCs stimulated with LPS (100 ng/mL). The protein expression were evaluated by Image J. ( F and G ) The protein expression of Nrf2 and GPX4 was evaluated by cell immunofluorescence (scale bar: 10 μm) and Image J. All data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001, n = 5.

Article Snippet: Tomatidine (Td, purity ≥98%) and lipopolysaccharide (LPS, purity >99%) were purchased from MCE (China).

Techniques: Gene Expression, Western Blot, Expressing, Immunofluorescence

Journal: Drug Design, Development and Therapy

Article Title: Tomatidine Alleviates Intervertebral Disc Degeneration by Activating the Nrf2/HO-1/GPX4 Signaling Pathway

doi: 10.2147/DDDT.S481714

Figure Lengend Snippet: The protective effect of Td on NPCs was reversed after Nrf2 was silenced. ( A ) The gene expression Nrf2 was detected by qPCR in the NPCs, treated with or without the administration of LPS for 24h. ( B and C ) After Nrf2 was silenced, Western blot analysis of ADAMTS-4, Aggrecan, Col-2, MMP3 and GAPDH expression in NPCs. Protein expression was assessed by Image J. ( D and E ) Col-2 expression was assessed by cellular immunofluorescence (scale bar: 10 μm) and Image J after Nrf2 was silenced. All data are presented as mean ± SD. ****P < 0.0001, n = 5.

Article Snippet: Tomatidine (Td, purity ≥98%) and lipopolysaccharide (LPS, purity >99%) were purchased from MCE (China).

Techniques: Gene Expression, Western Blot, Expressing, Immunofluorescence