Methanol extract of Smritisagara Rasa attenuates neurobehavior, neurochemical and stress indicators in rotenone-induced Drosophila model of Parkinson's disease

Smritisagara Rasa (SSR), a traditional Ayurvedic herbo-mineral formulation, has long been prescribed for the treatment of Parkinson’s disease (PD) since ancient times. Nevertheless, the underlying molecular mechanisms and treatment strategies remain unclear due to a lack of adequate experimental data. This study focused on formulating SSR and extracting its components using different solvent systems, including water and methanol. SSR, along with its water extract (SSR-WEX) and methanol extract (SSR-MEX), was tested for efficacy in mitigating PD phenotypes using a Rotenone (ROT)-induced Drosophila melanogaster PD model. SSR was prepared according to the Ayurvedic Formulary of India and subsequently characterized, followed by solvent extraction and phytoconstituent profiling. Different concentrations of SSR, SSR-WEX, and SSR-MEX were administered to control flies to evaluate their potential toxicity. Flies were then co-exposed to 500 μM ROT and 75 mg/kg diet of SSR, SSR-WEX, and SSR-MEX for seven days. Behavioral, neurochemical, and oxidative stress parameters were assessed, with pramipexole (PPX) used as a positive control. ROT exposure markedly impaired climbing ability, reduced dopamine and acetylcholinesterase (AChE) activity, and elevated oxidative stress markers, including malondialdehyde (MDA), protein carbonyl content (PCC), hydrogen peroxide, and nitric oxide. Co-treatment with SSR and its extracts significantly restored locomotor function, dopamine, and AChE levels, while enhancing antioxidant enzymes (glutathione-S-transferase and catalase) and reducing oxidative damage indices. Among the formulations, SSR-MEX demonstrated superior neuroprotective and antioxidative efficacy, comparable to the standard drug pramipexole. This effect is likely due to its enriched profile of flavonoids, coumarins, and phenolic acids. These findings substantiate traditional claims regarding SSR’s role in mitigating neurodegenerative symptoms and highlight SSR-MEX as a promising phytopharmaceutical candidate for managing PD-like pathologies through attenuation of oxidative stress and restoration of dopaminergic and cholinergic systems. Graphical abstract illustrates that preparation of herbo-mineral preparation of SSR and its extracts attenuates neurobehavior, neurochemical and stress indicators in ROT-induced Drosophila model of PD. • Smritisagara Rasa (SSR), a classical Ayurvedic herbo-mineral formulation, demonstrates promising neuroprotective potential in PD; however, its molecular mechanism remains largely unexplored due to limited experimental evidence. • SSR was formulated and characterized as per the Ayurvedic Formulary of India (AFI) and compared with its water (SSR-WEX) and methanol (SSR-MEX) extracts for efficacy using a rotenone (ROT)-induced Drosophila melanogaster model of PD. • Toxicity screening confirmed that SSR, SSR-WEX, and SSR-MEX were non-toxic at concentrations up to 75 mg/kg diet, ensuring the extracts safety for subsequent neurobehavioral and biochemical assays. • ROT-induced PD flies treated with SSR, SSR-WEX, SSR-MEX, and pramipexole (PPX) exhibited significant restoration of locomotor activity, DA levels, AChE, GST, and catalase enzyme activities, along with reduced oxidative stress markers such as MDA, PCC, H 2 O 2 , and NO. • Among all treatments, SSR-MEX showed the most pronounced therapeutic effect, exhibiting neurochemical and antioxidant restoration comparable to PPX, suggesting that its rich composition of bioactive phytochemicals synergistically counteracts dopaminergic degeneration and oxidative imbalance in PD. • The results scientifically validate the traditional Ayurvedic use of Smritisagara Rasa in neurological disorders, identifying SSR-MEX as a promising candidate for future drug development or adjunct therapy in PD management.