Arni Kusuma Dewi, Bambang Purwanto, Widjiati Widjiati, Viski Fitri Hendrawan
Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by impaired social interaction and stereotyped or repetitive behaviors. Prenatal exposure to valproic acid (VPA) is a well-established pharmacological risk factor for ASD phenotypes. This study evaluated the neuroprotective potential of fenugreek (Trigonella foenum-graecum) seed extract in a prenatal VPA-induced rat model of ASD. Pregnant Wistar rats were allocated into five groups (n = 6 per group): control (saline), VPA, VPA + fenugreek extract 100 mg/kg, VPA + fenugreek extract 200 mg/kg, and VPA + fenugreek extract 300 mg/kg. VPA (400 mg/kg) was administered as a single oral dose on gestational day (GD) 15. Fenugreek extract was administered orally once daily from GD11–GD20. Male offspring were assessed in a three-chamber social interaction test (Social Novelty Index; SNI) on postnatal day (PND) 27 and a Marble Burying Test (MBT) at PND35. Histological analysis (HandE) assessed viable neuron counts in the cerebral cortex, hippocampus CA1, and cerebellum, while myelin basic protein (MBP) immunohistochemistry quantified myelin density. The VPA group showed reduced social novelty preference and increased repetitive behavior compared with controls. Fenugreek extract at 200 mg/kg significantly improved the SNI score and reduced marble burying compared with the VPA group (p < 0.05). The 300 mg/kg dose showed stronger restoration of viable neuron counts, particularly in hippocampus CA1, although it did not significantly improve the SNI compared to the VPA group (p = 0.17). MBP intensity increased in the 200 and 300 mg/kg groups, indicating improved myelination. Fenugreek seed extract exhibits neuroprotective potential against prenatal VPA-induced ASD-like phenotypes, with 200 mg/kg showing the most consistent behavioral benefit and 300 mg/kg showing stronger histological recovery. Further studies are required to elucidate dose-dependent mechanisms and confirm bioactive compound contributions. This study supports SDG 3 (Good Health and Well-being) by exploring plant-based neuroprotective strategies for ASD. It also aligns with SDG 9 (Industry, Innovation and Infrastructure) through its potential to inspire future development of phytopharmaceuticals. © 2026 by the authors. Licensee ResearchersLinks Ltd, England, UK.
Faculty of Medicine, Universitas Airlangga, Campus A, Jl. Prof. Dr. Moestopo No.47, Pacar Kembang, East Java, Surabaya, 60132, Indonesia; Department of Physiology and Medical Biochemistry, Faculty of Medicine, Airlangga University, (Campus A) Prof. DR. Moestopo No.47 Street Pacar Kembang, East Java, Surabaya City, 60132, Indonesia; Departement of Veterinary Medicine, Faculty of Veterinary Medicine, Airlangga University, (Campus C) Dharmahusada Permai No.1 Street, Mulyorejo, East Java, Surabaya City, 60115, Indonesia; Department of Veterinary Reproduction, Brawijaya University, Puncak Dieng Street, Kunci-Kalisongo, East Java, Malang, 65151, Indonesia
