Abstract
AbstractDicliptera is a genus belonging to family Acanthaceae. Dicliptera bupleuroides is a small perennial herb commonly known as roxburg's foldwing. It is locally used in folkloric medicines for the treatment of different diseases and as a general tonic for wound healing. The current investigational study was to optimize solvent extraction protocol, to identify quantitative and qualitative biochemical composition, antioxidant and cytotoxic activities of D. bupleuroides and to characterize the gene associated with biosynthesis of flavonol. The plant of D. bupleuroides was collected from district Abbottabad and separated into its four parts including leaf, stem, flower, and root. To extract the biochemical compounds, five different solvents were used such as methanol, acidified methanol, 80 % methanol, ethyl acetate, and n‐butanol. Total phenolic, flavonoids, flavonol, flavanol (catechin), and anthocyanin contents were determined and antioxidant potential of the extracts was evaluated using various antioxidant assays. The methanolic extracts of D. bupleuroides showed promising antioxidant potential towards DPPH, ABTS, H2O2, ferric, phosphomolybdate and hydroxyl radical scavenging capabilities.The extracts obtained from the solvents mentioned above were tested for cytotoxicity using the 3T3 cell line by using mouse fibroblast cells. The results indicated that none of the extracts demonstrated any toxic effects on the cells, suggesting that they are non‐toxic. The RNA was extracted and cDNA was synthesized using reverse transcriptase enzyme. Degenerate primers for dihydroflavonol reductase (DFR) gene were synthesized to amplify the desired product. The high quality of cDNA amplification showed the presence of the DFR gene in the Dicliptera bupleuroides leaf for the first time with a product size of 419 bp respectively. The DFR gene isolated from Dicliptera bupleuroides is involved in flavonol biosynthesis regulation. In addition, Glide molecular docking results proposed that the constituents within the n‐hexanic extract of Dicliptera bupleuroides not only furnish an updated approach to better understand the antioxidant potential of its various constituents to interact with an antioxidant target protein (PDB ID:3MNG) but also ensure promise in addressing these potential therapeutic modalities. The active compounds of D. bupleuroides could be suitable for pharmaceutical industries to develop stress related drugs.
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