Elicitation increased chromones accumulation and anti-inflammatory activity in Harrisonia perforata (Blanco) Merr. cell cultures

Abstract

Harrisonia perforata is an essential ingredient in Thai traditional medicine's Ha-Rak herbal preparation, traditionally requiring equal proportions of roots from five plants, raising sustainability concerns regarding sourcing from natural habitats. This study aimed to establish an in vitro callus and cell suspension culture system of H. perforata for sustainable production of two bioactive chromones—O-methylalloptaeroxylin and perforamone D—and to evaluate their anti-inflammatory potential. Optimal callus induction and cell suspension growth were achieved using Murashige and Skoog (MS) medium supplemented with 1.0 mg/L TDZ and 0.5 mg/L NAA. Treatment of cell suspension cultures with elicitors significantly enhances chromone accumulation. Maximum O-methylalloptaeroxylin (60.93 ± 0.62 µg/g DW; 19.2-fold increase) was recorded after elicitation with 10 µM salicylic acid (SA) for 3 days, while perforamone D reached its highest level (18.74 ± 1.17 µg/g DW; 1.7-fold increase) following treatment with 100 mg/L yeast extract (YE) for 3 days. Anti-inflammatory activity, assessed by nitric oxide inhibition in LPS-stimulated RAW 264.7 macrophages, correlated significantly with perforamone D concentration (r = 0.387, p < 0.01). The established in vitro culture system demonstrates the potential for sustainable production of these valuable compounds. However, further optimization is required for practical large-scale applications.