Previous studies showed that augmentation of the autophagy-lysosome process may be beneficial for many neurodegenerative diseases including Parkinson's disease (PD), in which abnormal accumulation of aggregated proteins and mitochondrial dysfunctions are considered as crucial pathogenesis. Recently, several studies have advocated the neuroprotective effects of intralipid in models of numerous neurodegenerative diseases. In this study, seventy "C57BL/6" mice divided into seven groups were used to evaluate the neuroprotective effects of autophagy enhancement using metformin and intralipid through neurobehavioural, a histopathological and immunohistochemical examination of rotenone-exposed mice brain. The mice were administrated rotenone (2.5 mg/kg intraperitoneally), metformin (100 mg/kg/day orally) and intralipid 20% (15 mL/kg intraperitoneal) daily for 4 weeks. Intralipid significantly recovered the damaged brain tissue and alleviated rotenone-induced brain damage. Furthermore, the co-administration of metformin and intralipid against rotenone produced more therapeutic efficiency. Intralipid exerted protection against rotenone-induced neuro-injuries to some extent through the autophagy process. The pharmacological induction of autophagy by intralipid may represent a novel therapeutic strategy as a disease-modifier in neurodegenerative diseases.