Phytochemical Profiling of Helianthus annuus L. seed and Investigation of Developmental Toxicity in Zebrafish Embryos with Anti-Osteoporotic Evaluation in Adult Models

Abstract

This study investigates the pharmacological potential of Helianthus annuus L. (sunflower) defatted seed cake, a discarded residue after oil extraction. Rich in phytochemicals like flavanones and polyphenols, this material is shown to exhibit various activities, including anti-inflammatory and antioxidant properties. The extract was evaluated for its phytochemical analysis, antioxidant and anti-inflammatory activities. The primary focus was evaluating its anti-osteoporotic activity, given the lack of scientific data on this specific property. Osteoporosis was induced in a zebrafish model using Ferric ammonium citrate (FAC), and promising results were observed with extract administration (100 and 200 µg/ml) in preventing osteoclast formation and enhancing osteoblast formation. Furthermore, the embryo toxicity of the extracts on developing zebrafish embryos was assessed at various concentrations (3.125-1000 µg/ml) over 24-96 hours, demonstrating its safety as per OECD guideline. The presence of bioactive constituents revealed phenolic and flavonoid content in the extract. LC-MS/MS profiling further confirmed the presence of several polyphenolic compounds. In vitro antioxidant assays and anti-inflammatory protein denaturation assays demonstrated promising effect. Developmental toxicity testing demonstrated the LD50 lies in between 12.5 and 3.125 µg/ml. In vivo, pretreatment with FAC (250 μg/ml) significantly attenuated a quite improvement in bone resorption and bone density The findings suggest that the anti-osteoporotic effects of Helianthus annuus L. may stem from its anti-inflammatory and antioxidant properties, highlighting its potential as a natural therapeutic agent for bone density improvement and supporting further investigation into its active constituents.