Comparative Profiling of Antioxidant Mechanisms in Averrhoa Species Using Integrated In Vitro and In Silico Approaches

Woro Anindito Sri Tunjung, Ane Tefvy Styorini, Hilman Wahyu Pratama, Abimanyu Dimas Cahyo Setyanto, Risqi Rahmawati, Wulan Usfi Mafiroh, Andrea Maharani, Frisca Damayanti, Nor Ismaliza Mohd Ismail

Abstract

Reactive oxygen species (ROS) contribute to oxidative stress and are associated with the development of chronic diseases, increasing the need for effective natural antioxidants. Although Averrhoa species are known to contain bioactive compounds with antioxidant potential, their underlying molecular mechanisms and interspecies differences remain poorly understood due to the lack of integrative studies. In this study, we investigated the antioxidant activity and bioactive compounds of four Averrhoa species (Averrhoa bilimbi, Averrhoa carambola, Averrhoa dolichocarpa, and Averrhoa leucopetala) leaves using in silico and in vitro approaches. Leaf powder of Averrhoa spp. was macerated in 70% ethanol, then a GC-MS profiling identified 18-24 compounds in A. dolichocarpa, A. bilimbi, and A. leucopetala, compared to only four compounds in A. carambola. Subsequent network pharmacology analysis prioritized vitamin E, threonine, squalene, butanal, and neophytadiene as key bioactive compounds based on their target connectivity and association with antioxidant-related pathways. Molecular docking revealed favorable binding of vitamin E toward superoxide dismutase (SOD) and Kelch-like ECH-associated protein 1(Keap1), suggesting potential roles in antioxidant enzyme modulation and Nrf2 pathway activation. Enzyme assays revealed that A. dolichocarpa exhibited the highest superoxide dismutase (SOD) (170.25 U/min/gFW) and catalase (CAT) (2.05 U/min/gFW) activities, A. leucopetala showed the highest ascorbate peroxidase (APX) activity (2.22 U/min/gFW), whereas A. bilimbi showed the strongest radical scavenging activity (IC₅₀ = 15.24 µg/mL). These findings indicate that antioxidant activity in Averrhoa is species-dependent, involving enzyme-mediated defense and direct radical scavenging. Overall, this integrative approach provides comprehensive insight into their antioxidant mechanisms and highlights their potential for nutraceutical and pharmaceutical applications.

Keywords

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DOI: 10.14416/j.asep.2026.08.016

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