Unraveling the Anti-Multiple Myeloma Activity of Alisol B 23-Acetate and Its Impact on the P38MAPK Pathway Through Integrated Analysis

Alisol B 23-acetate is a naturally occurring triterpenoid compound derived from the rhizome of the traditional Chinese medicinal herb Alisma orientale,which possesses a variety of physiological activities, including anti-cancer effects.However, its effects on human multiple myeloma (MM) and the underlying mechanisms remain to be elucidated. In this study, we employed an integrated analysis of mRNA, miRNA, and network pharmacology to investigate the role of Alisol B 23-acetate in MM and subsequently validated our findings with cellular experiments.Our data demonstrate that Alisol B 23-acetate treatment significantly suppresses MM cells via the P38MAPK signaling pathway. Specifically, the CCK-8 assay revealed that Alisol B 23-acetate inhibits the proliferation of MM cells, with an IC50 value of 14.24 µM after a 24-hour treatment. Cell cycle analysis indicated that Alisol B 23-acetate treatment increased the percentage of MM.1S cells in the G0/G1 phase (resting/before DNA synthesis) and decreased the proportion in the S/G2 phase (DNA replication/after DNA synthesis). Apoptosis assays showed that Alisol B 23-acetate significantly enhanced apoptosis in MM.1S cells, with the apoptotic cell population increasing to 57.3%. Quantitative real-time PCR (qRT-PCR) results indicated that Alisol B 23-acetate treatment downregulated the expression of the anti-apoptotic gene Bcl2 and upregulated the expression of the pro-apoptotic gene Bax in MM cells, while the expression of the P38 gene remained largely unchanged. Western blotting analysis further confirmed that Alisol B 23-acetate treatment reduced the expression of the anti-apoptotic protein Bcl2 and increased the expression of the pro-apoptotic protein Bax in MM cells, with a concomitant decrease in P38 protein phosphorylation.These findings suggest that Alisol B 23-acetate induces apoptosis in MM cells by inhibiting the phosphorylation of P38, a key target in the P38MAPK signaling pathway.