Ameliorative effect of AYUSH formulation against scopolamine induced learning and memory impairment in rat via attenuation of oxidative stress, neuroinflammation, acetylcholinesterase inhibition and Aβ plaques.

ETHNOPHARMACOLOGICAL RELEVANCE: Itrifal Muqawwi Dimagh (IMD) is a traditional polyherbal Unani formulation known to support cognitive function. While it is traditionally believed to have neuroprotective and memory enhancing effects, systematic scientific studies validating it's therapeutic efficacy and the underlying mechanism of action are still limited.

AIM OF THE STUDY: The objective of the study was to determine the neuroprotective potential of IMD against scopolamine induced learning and memory impairment.

MATERIALS AND METHODS: The quality control studies include the simultaneous quantification of gallica acid, and ellagic acid using developed HPTLC method. The dose-dependent neuroprotective potential of IMD were investigated in scopolamine induced model in albino Wistar rats at three doses levels to assess dose-dependent effects (86 mg/kg, 172 mg/kg, and 344 mg/kg). Neurobehavioral parameters were assessed using Morris water maze test and spontaneous alternation behaviour. The biochemical estimation using acetylcholinesterase inhibition, amyloid beta (Aβ) estimation, oxidative stress, and anti-inflammatory markers were also carried out followed by histological and immunohistological analysis. Furthermore, in-vivo pattern recognition analysis of IMD metabolites in serum, was done to gain further insights.

RESULTS: The present investigation represents the first report on IMD in scopolamine induced learning and memory impairment model and tentative metabolomic profiling. Treatment with IMD significantly reduced escape latency and transfer latency, and increased percentage dwell time as compared to the scopolamine group, in a dose-dependent manner. Treatment with medium (IMDM) and high (IMDH) doses of IMD, significantly attenuated the elevated biochemical parameters, restored the antioxidant level (SOD), and regulated the expression of proinflammatory cytokines (TNF α, and IL-6) in brain tissue showing comparable results to that of standard. The neuroprotective effects of IMD were supported by its ability to inhibit acetylcholinesterase activity and mitigate amyloid beta accumulation, comparable to that of standard. The in-vivo pattern recognition analysis of IMD, revealed presence of 27 metabolites, out of which 20 metabolites were identified and seven metabolites were unidentified at different time intervals.

CONCLUSION: IMD supported overall brain function through multiple mechanisms including anti-oxidant, anti-inflammatory, AChE inhibition and anti-amyloidogenic pathways. While the current study suggests a possible neuroprotective effect of IMD, additional studies are needed to substantiate these findings and determine their broader applicability.