Multi-Components in Modified

Objective: To identify active components of Modified Methods: Data mining of EC-related target genes was performed. Protein-protein interaction (PPI) network and core target analysis were conducted, followed by analysis of the relevant autophagy pathway. Active compound docking with key targets was performed. Medicated serum of MTD was prepared from Sprague-Dawley (SD) rats that received intragastric administration of low (LD), medium (MD), and high-dose (HD) MTD. Concentrations of the top active compounds in the serum were measured with HPLC. EC TE-1 cells were cultured in medicated serum. Autophagic acidic vesicles and autophagosomes were observed. Protein expression of autophagy-related and Akt/ mTOR pathway molecules was assayed. Protein expression of LC3 was determined. mRNA expression of Beclin-1 was measured. Results: The top four active compounds were quercetin, baicalein, luteolin, and β-sitosterol. The top four targets were TP53, CTNB1, EGFR, and Akt. Akt and mTOR were intersecting molecules involved in the EC autophagy pathway. The top compounds co-targeted Akt equally. Concentrations of the top compounds in the medicated serum accounted for 14-15%. MD increased the fluorescence intensity of acidic autophagy vesicles. LD and MD promoted an increase in autophagosomes. HD strongly affected Beclin-1, LC3, Akt, and mTOR. MD inhibited protein expression of p62 but promoted LC3 fluorescence intensity. These three doses had equal effects on mRNA expression of Beclin-1. Conclusion: MTD specifically promotes EC autophagy depending on multiple components and specifically targets Akt/mTOR signaling pathway. MTD shows clear activity in EC cell autophagy regulated by the Akt/mTOR pathway.