Fig. 7
AS-IV enhances HSC proliferation via the AMPK/PGC1α signaling pathway. K562 cells were subjected to treatment with Compound C (5 μM), Compound C (5 μM) combined with AS-IV (10 μM), or AS-IV (10 μM) for a duration of 5 days. A Molecular docking shows the binding ability between AS-IV and its core target (AMPKα); B–C DARTS assays demonstrated dose-dependent binding of AS-IV to AMPKα in K562 cells. Treatment with Streptomyces protease (1:1000) at 40 °C for 10 min; D The lysates of K562 cells were treated with AS-IV (200 μM) for 1 h, followed by the addition of streptomycin (Pronase E) at various concentrations (1:500, 1:1000 or 1:1500), and incubated at 40 °C for 10 min. AMPKα content was assessed using western blot analysis; E CETSA analysis of AMPKα degradation damage under different temperatures. The histogram shows the relative density of AMPKα to GAPDH; F CCK-8 assay for K562 cell proliferation; G Flow cytometry analysis was conducted to assess the mitochondrial membrane potential in K562 cells; H ATP assay kit to detect the concentration of ATP produced by K562 cells; I–K The expression of PGC1α and p-AMPKα was detected by WB. L The bar graph shows the viability of K562 cells after AMPKK45R gene mutation, treated with 10 μM AS-IV; M Flow cytometry was used to detect the changes of mitochondrial membrane potential in K562 cells with AMPKK45R gene mutation. Loading or not loading AS-IV intervention; N ATP assay kit was used to detect the level of ATP production in K562 cells with AMPKK45R mutation; O–Q Western blot was used to detect the changes of p-AMPKα and PGC1α in K562 cells with AMPKK45R mutation. Data represent the mean ± standard deviation of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus the corresponding control groups