Exploratory evaluation of the neuroprotective potential of Alfalfa in an AlCl₃-induced autism-like zebrafish model.

Autism spectrum disorder (ASD) is a neurodevelopmental condition with compromised social interaction, communication deficiency, and stereotypical behaviour. AlCl₃ causes neurotoxicity that mimics autistic-like behaviours in model systems. Alfalfa (Medicago sativa) is a medicinal plant with high phytonutrient quality and is documented to display antioxidant and anti-inflammatory activities. The anti-inflammatory and neuroprotective activity of Alfalfa was assessed against AlCl₃-induced autism-like behaviour in adult zebrafish in the current study. The experiment consisted of 12 zebrafish per group in which on Day 0; AlCl₃ was given through immersion to cause autism-like effects. Alfalfa extract doses of 45 µL and 90 µL were given as treatment, whereas Risperidone served as the standard drug for 28 days. Neurobehavioral tests such as Novel object recognition test (recognition memory task), Novel tank Test (anxiety), and Light-Dark Preference Test (learning and memory deficit) were performed, and biochemical indicator such as content, LPO, reduced glutathione GSH, nitrite content, AChE activity, CAT activity, and neurotransmitters GABA and Glutamate, and inflammatory cytokines TNF-α, and IL-1β were estimated at the end of the study. Histopathological examination was also carried out at the end of the treatment. Alfalfa enhanced behaviour, restored neurotransmitter balance, minimized oxidative stress, regulated inflammation responses, and exerted protective effects on neural tissue in zebrafish with AlCl₃-induced autistic behaviours. These results illustrate the therapeutic implications of Alfalfa as a protective and anti-inflammatory drug in autism spectrum disorder treatment.