Cytotoxicity of Ethanolic and Methanolic Extracts of Medicago sativa L. (Alfalfa) on K562 Myeloid Cancer Cell Lines

Background & Objectives: For centuries, the alfalfa (Medicago sativa L.) plant has been recognized for its versatile and active role in treating various diseases. Not only has it been utilized as a therapeutic agent, but it has also been served as a dietary component for both animals and humans. Given the distinctive attributes of this plant in ethnopharmacology, this study aimed to investigate the effects of ethanolic and methanolic extracts of M. sativa L. on the K562 myeloid cell line under in vitro conditions. Materials & Methods: The phytochemical composition of M. sativa L. was determined through Gas Chromatography-Mass Spectrometry (GC-MS) analysis. Cell viability was assessed using the MTT assay, wherein K562 cells were subjected to varying concentrations (50–100 μg/mL) of methanolic and ethanolic extracts over 24, 48, and 72- hour intervals to determine the IC50. Subsequently, the most promising IC50 result was employed in flow cytometry (Flow Jo Software) analysis. Results: Active constituents identified included phytol, phenol, linolenic acid, and glycine. Statistical analysis revealed a time-dependent but not dose-dependent effect. It was noteworthy that the IC50 for the methanolic extract after 72 hours was 9.45 μg/mL, whereas it was 19.3 μg/mL for the ethanolic extract. Flow cytometry analysis indicated that the methanolic extract caused 49.16% and the ethanolic extract caused 15.42% of cell death. Conclusion: The results demonstrated that the ethanolic extract of alfalfa is more effective than the methanolic extract on the K562 cell line. Therefore, M. sativa L. potential application in myeloid cancer therapy can be investigated in more details