Evaluation of the cytotoxic potential of Annona muricata Linn leaf extract against N2a-neuroblastoma cells

Abstract

Background: Plants are rich in bioactive compounds with various pharmacological properties, and they can be used effectively to treat various diseases. The ability of Annona muricata, preventing progression of cancer cells by initiating programmed cell death, was focused in the present study. Objective: The objectives of this study were to determine the antioxidant and anticancer potential of A. muricata ethanolic extract (AME) of leaves. Materials and Methods: The antioxidant potential of AME was determined by 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging activity, 2,2’-azinobis(3-ethyl-benzothiozoline)-6-sulfonic acid ammonium salt (ABTS) radical cation scavenging activity, ferric reducing antioxidant power (FRAP) assay, phosphomolybdenum assay, and the cytotoxic potential of AME over N2a cell lines (neuroblastoma cells) was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, lactate dehydrogenase (LDH), intracellular reactive oxygen species (ROS), and measurement of mitochondrial membrane potential (MMP) assay. Results: The 50% scavenging ability of the extract in DPPH assay was found to be 14 μg/mL. The ABTS radical scavenging potential was measured to be 10507.3 mg/g of extract. The FRAP was found to be 10.38 mM Fe(II)E/mg extract. The effective phosphomolybdenum reduction was measured at a concentration of 53.33 mg ascorbic acid equivalents/g extract. The in vitro cytotoxicity of AME with escalating concentrations (5–85 μg/ml) was first screened, and it was shown to inhibit N2a cells with IC50 value of 55 μg/ml. A significant increase in cytosolic enzyme LDH confirms the cell membrane damage. The extent of cell damage was confirmed by a significant increase in the level of intracellular ROS and a decrease in MMP. Furthermore, to characterize and identify the major active constituents present in AME, Fourier-transform infrared and gas chromatography-mass spectrometry analysis were carried out. Conclusion: The present results demonstrated that AME could be a promising candidate for developing anticancer agents which helps further to combat neuroblastoma with an effective treatment strategy