Phytochemical Analysis (LC-MS) of Azadirachta Indica Ethanolic Extract, Antioxidant, Anticancer, in Vitro Wound Healing Activity, and Immunomodulatory Effects of Azadirachta Indica Ethanolic Extract

Abstract

This study presents a comprehensive investigation of the medicinal attributes of the ethanolic extract obtained from Azadirachta indica (A. indica) leaves collected from the Jamia Salafiya Pharmacy College campus in Malappuram District, Kerala, South India. The plant was meticulously identified by Dr. Samuel Thavamni B, a plant pharmacognosy specialist. The ethanolic extraction process was performed using the Soxhlet method. Phytochemical analysis was performed using liquid chromatography-LC-MS (liquid chromatography-mass and Mass spectroscopy). The in vitro antioxidant, MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay, Apoptosis, and Immunomodulatory activity of ethanolic A. indica were evaluated. The extraction resulted in a semi-solid, dark-coloured extract with a yield of 8.67%. Phytochemical analysis using LC-MS identified key constituents such as octadecanoic acid, tetradecanoic acid, caryophyllene, 1,4-Eicosadiene, and triacontanoic acid, methyl ester, known for anti-inflammatory, antioxidant, and antimicrobial properties. In vitro antioxidant assays revealed a concentration-dependent inhibitory effect, with the A. indica ethanolic extract demonstrating substantial antioxidant capacity (77.81% at 100 µg), comparable to that of ascorbic acid (97.22%). However, the MTT assay indicated a concentration-dependent decrease in cell viability, emphasizing the need for caution owing to potential cytotoxic effects. Acridine orange/ethidium bromide staining provided insights into the induction of apoptosis in MCF-7 cells, highlighting the beneficial effects of the extract on cellular morphology. The scratch wound healing assay suggested the potential of the extract to promote fibroblast migration, implicating its role in enhancing wound healing. Immunomodulatory activity assessment revealed concentration-dependent modulation of immune responses, with the extract demonstrating a notable immunomodulatory effect of 77.81% at 100 µg. These findings shed light on the medicinal potential of A. indica, emphasizing the need for further research to elucidate the underlying mechanisms. The diverse chemical composition and multifaceted effects of the extract, from antioxidant and cytotoxic activities to immunomodulation and wound healing promotion, underscore its significance in pharmaceuticals and herbal remedies.