Journal: Frontiers in Medicine
Article Title: Network pharmacology, machine learning, and experiments uncover β-sitosterol targeting HSP90AA1 in medicinal-edible black soybean against aging
doi: 10.3389/fmed.2026.1749856
Figure Lengend Snippet: HSP90AA1 mediates β-sitosterol’s anti-photoaging effects. (A) HSP90AA1 expression upregulated by UVA and further enhanced by β-sitosterol (mRNA). (B,C) Co-treatment with HSP90AA1 inhibitor IPI-504 reverses β-sitosterol-induced stabilization of pro-survival BCL-2 and p53 (mRNA). (D) ROS scavenging capacity of β-sitosterol blocked by IPI-504 (DCFH-DA assay). (E) Cell cycle analysis showing β-sitosterol alleviates UVA-induced G1 arrest, abrogated upon HSP90AA1 inhibition. (F) Western blot analysis confirming efficient knockdown of HSP90AA1 protein by specific siRNA (siHSP90AA1). (G) HSP90AA1 knockdown abolishes β-sitosterol’s stabilizing effect on p53 and BCL-2 mRNA levels under UVA stress. (H) HSP90AA1 knockdown blocks the ability of β-sitosterol to scavenge UVA-induced ROS. (I) ROS scavenging capacity of Tam (1 μmol/L). (J) Cell cycle analysis showing Tam alleviates UVA-induced G1 arrest. Data shown as mean ± SEM, n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001.
Article Snippet: The cells were maintained at 37°C in an atmosphere containing 5% CO2. β-sitosterol (Tokyo Chemical Industry, Tokyo, Japan) was dissolved in the culture medium and used at a concentration of 25, 50 or 100 μg/mL for 24 h treatment.
Techniques: Expressing, DCFH-DA Assay, Cell Cycle Assay, Inhibition, Western Blot, Knockdown