Extraction, Phytochemical profile and Neuroprotective Activity of Phyllanthus emblica Fruit Extract Against Sodium Valproate-Induced Postnatal Autism in BALB/c Mice via modulation of 5-HT1D, 5-HT2A, D2 receptor protein, oxidative stress and inflammatory biomarkers

Abstract

The aim of this study was to evaluate the neuroprotective effect of ethyl acetate fraction of amla (EAFA) extract against sodium valproate or valproic acid (VPA)-induced autism in postnatal BALB/c mice and explore its mechanism of action on serotonergic and dopaminergic systems as well as oxidative stress. In this in vitro study, EAFA extract had a higher total antioxidant activity than the 70% methanol amla extract by the ferric-reducing antioxidant power test (FRAP), and the EAFA extract showed more potent NO free radical scavenging activity than the 70% methanol amla extract. In this study, phytochemical constituents were identified by UPLC-Q-TOF-MSE from the EAFA extract, as a result, major compounds were quantified by using UPLC-MS/MS analysis. In the present in vivo study, treated with EAFA extract 100 mg/kg BW from postnatal day13 (PND13) to PND 40 against VPA-induced postnatal autism (VPA single dose 400 mg/kg BW s.c. PND 14). All experimental groups were subjected to batteries of behavioural parameters from PND 23 to PND 40, and all animals were sacrificed on PND 41. The brain was isolated for further analysis, such as biochemical analysis, pro-inflammatory markers, histopathological analysis, IHC and western blot analysis. The VPA-alone group showed significantly increased the disease hallmarks in autism-like disease markers and pathogenesis compared to the control group. Our study highlights the protective effect of EAFA extract on improving behavioural alterations, significantly restore the anti-oxidative enzymes such as GST, GR and reduced the MDA and NO levels. Furthermore, the EAFA treated group significantly lowering the pro-inflammatory markers (IL-1β and TNF-α), and the expression of up-regulated 5-HT1D, 5-HT2A, and D2 receptor proteins. Based on histopathological studies, the percentage of neuronal injury in the EAFA-treated group was reduced. According to this study's findings, EAFA extract's ameliorating activity is due to its anti-oxidant and neuroprotective efficacy against VPA-induced autism.