Evaluation of anticancer, immunomodulatory and anti-inflammatory potential of antioxidant rich Anzia ornatoides, a lichen species from eastern Himalayan region

Abstract

The lichen Anzia ornatoides was identified by its morpho-anatomical and chemical features. The hexane, diethyl ether, ethyl acetate, methanol, and water extracts of the lichen were assessed for their antioxidant capacities. The presence of various phyto-compounds of the methanol extract were run through a GC–MS and identified three major compounds viz. methyloxyolivetol, imidazole 2-t‑butyl‑1,4-dimethyl-5-phenyl and benzoic acid 2,4-dihydroxy-3,6-dimethyl-methyl ester. The crude extraction of a lichen sample was carried out by Soxhlet apparatus. Total phenolic content was determined by Folin-Ciocalteau assay while flavonoid content was estimated using aluminium chloride assay. The antioxidant activity of the extracts was analysed by phosphomolybdenum assay, FRAP, DPPH, ABTS, and lipid peroxidation inhibition. To see the immunomodulatory and anti-inflammatory potential of the extracts, cell viability assay of macrophage was studied. In in-vitro anticancer activity, the extracts were explored using cancer cell line of different organ and tissue origin it includes PC-3, OVCAR-3, hep-G2, h-1299, HeLa, and MCF-7. Apoptotic potential is studied using the OVCAR-3 cell line. In-vitro cytotoxicity assay was conducted with the non-cancer cell lines HEK-293, L-929, hMSCs, and MCF-10A. Using mouse model, in-vivo animal toxicity was also carried out. The lichen extracts showed a good amount of phenolic and flavonoid content and high antioxidant potential. Based on the findings of the experiment, methanolic extracts of A. ornatoides showed higher immunomodulatory, anti-inflammatory and anticancer capabilities. In an in-vitro cytotoxicity test against the cancer cell line IC50 value ranged from 38 to 78 µg/ml. Based on in-vivo toxicological evaluation and histopathological analysis, no significant toxicity was detected in-vitro cultured non-cancerous cells or in the liver or kidney tissue of mice.