Eun Jin Seo, Soyoung Park, Eungyung Lee, Yang Hoon Huh, Ye Eun Ha, Gabor J. Tigyi, Taesung Jeong, Il Ho Jang, Jonghyun Shin. IJSC 2024;17:330-6. https://doi.org/10.15283/ijsc23105

The seeds of Impatiens balsamina L. have been traditionally used in East Asian medicine and are known to contain bioactive compounds with antioxidant properties. However, studies focusing on seed-derived callus remain limited. This study aimed to comparatively evaluate the antioxidant activities and lipid accumulation-inhibitory effects of 70% ethanol extracts from seeds (IB) and seed-derived callus (IBC) of I. balsamina. Callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). Antioxidant activities were evaluated using DPPH radical scavenging, superoxide anion scavenging, deoxyribose-based hydroxyl radical scavenging, DNA nicking, lipid peroxidation, and relative electrophoretic mobility (REM) assays, along with the determination of total phenolic, flavonoid, and tannin contents. Cell viability and lipid accumulation were assessed in FFA-treated HepG2 cells. In silico network and transcription factor (TF) enrichment analyses were performed to explore underlying mechanisms. Callus induction was most effective at 1 mg/L 2,4-D. Both IB and IBC exhibited antioxidant activities across all assays, with IB showing higher activity and greater phytochemical content than IBC. Both extracts reduced lipid accumulation in FFA-treated HepG2 cells at non-cytotoxic concentrations. Network analysis identified enrichment in pathways related to oxidative stress, inflammation, and lipid metabolism, and TF enrichment analysis identified NFKB1 and ATF3 as major upstream regulators. Both IB and IBC exhibited antioxidant activities across multiple in vitro assays, with IB showing higher activity attributable to its more complex phytochemical content. The lipid accumulation-inhibitory effects observed in FFA-treated HepG2 cells suggest a potential association between antioxidant capacity and lipid regulation, although the underlying mechanisms remain to be experimentally validated. Seed-derived callus may serve as a useful in vitro model for studying plant-derived bioactive compounds, pending further optimization.