Systems-Level Analysis of 13-Docosenamide Reveals its Regulatory Role on PPAR, AMPK, and Insulin Pathways in T2DM: In vitro and In silico Approaches
Abstract
Type 2 diabetes mellitus (T2DM) is a multi-faceted metabolic disease marked by the malfunctioning of fat tissue, improper lipid metabolism, and the failure of insulin-related signaling pathways. Through the secretion of adipokines and the activity of lipid-modulating enzymes, adipose tissue establishes itself as the central endocrine regulator of metabolic homeostasis and thus becomes a potential therapeutic target. The current study took advantage of a combined approach using phytochemistry, biochemistry, and systems biology to shed light on the mechanistic contribution of Hygrophila auriculata and its bioactive constituent in the regulation of T2DM-associated pathways. Gas chromatography was used for profiling of the ethanolic extract, and it successfully identified seventy-three phytoconstituents, among which 13-docosenamide was found to be the most abundant. The compound was obtained at a high purity level through preparative HPLC (RT 3.801 min, 99.997%) and was structurally confirmed as (Z)-13-docosenamide through GC–MS, FTIR, and UV–Vis analyses. The functional validation showed strong α -glucosidase inhibition (81.34% at 500 µg/mL; IC₅₀ = 71.41 µg/mL) and moderate α-amylase inhibition which pointed out the suppressive effect of postprandial hyperglycemia. The low toxicity (IC₅₀ = 445.3 µg/mL) revealed by cytotoxicity evaluation in 3T3-L1 adipocytes reinforced the biological safety of the compound. Network pharmacology analysis pointed out 14 common targets that…
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