Abstract
Roylea cinerea (D. Don) Baill. (Lamiaceae) is an indigenous plant of Western Himalayas, and has been used by the native population for the treatment of various diseases such as fever, malaria, diabetes, jaundice, and skin ailments. However, limited proportion of pharmacological and toxicological information is available on the bioactive properties of this plant. Therefore, the present study was designed to explore the anti-oxidant and anti-proliferative activities of Roylea cinerea. Methanolic extracts of leaves and stem of Roylea cinerea were prepared through maceration procedure and evaluated for the antioxidant activity using hydrogen/electron donating and hydroxyl radical scavenging assay. Significant antioxidant activity was observed for the methanolic extract of leaves in DPPH (EC50 239 µg/ml), molybdate ion reduction assay (29.73 µg ascorbic acid equivalent/mg dry weight of extract) as well as in plasmid nicking assay. Anti-proliferative and apoptotic activity in L6 rat skeletal muscle cell line was done using in vitro assays, i.e., MTT, Lactate dehydrogenase, mitochondrial membrane potential assay along with phase contrast, confocal, and scanning electron microscopy. The methanol extract of leaves and stem inhibited the growth of L6 cells with IC50 value of 69.41µg/ml and 124.93 µg/ml, respectively, and the lactate dehydrogenase activity was 20.29% and 0.3%, respectively. Cell cycle analysis by flow cytometry exhibited the arrest of cells in G1 and sub-G1 phase by methanolic leaves extract. Furthermore, the results of microscopic and docking analysis strengthened the observation made in the present study regarding the apoptotic mode of cell death in the L6 cell line. The in vitro findings of our studies revealed that the bioactive ingredients present in the methanolic extract of leaves and stem of Roylea cinerea have the anticancer potential. Further in vivo studies are needed to verify the in vitro results.
Highlights
Reactive oxygen species (ROS) are normally produced in the body from the mitochondria and are often termed as ‘redox messengers'
Medicinal plants demonstrating higher antioxidant activity have been reported to contain a high amount of phenolic compounds
The results indicated the increase in apoptotic cells and cell cycle arrest at G0/G1 phase in treated cells as compared to the nontreated cells which may be attributed to DNA damage mediated p53 activation to check further cell proliferation (Figure 4E) (Khan et al, 2019)
Summary
Reactive oxygen species (ROS) are normally produced in the body from the mitochondria and are often termed as ‘redox messengers'. The ROS form an integral part of various intracellular signaling pathways. Enhanced exposure to xenobiotics and oxidative stress generate prodigious levels of ROS, which in the absence of antioxidant defense pathways can attack cell membrane and alter the structure of cellular macromolecules, protein functioning and may cause mutations in cellular DNA. Several studies have confirmed the relationship between elevated levels of ROS and carcinogenesis (Weinberg, 1989). The multistep process of carcinogenesis commences through disturbed homeostasis between deviant proto-oncogenes activation and suppression of tumor suppressor genes with critical pathways and biomarkers (Rashid, 2017). Cancer chemoprevention edges on unraveling the potent cost-effective anticancer agents that can influence cellular transformations in the early stages. Despite numerous beneficial effects of synthetic drugs, naturally occurring phytochemicals are preferred as potential anticancer therapies considering the lesser toxicity and fewer side effects
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