Rannasangpei and its constituent crocin-1 ameliorate valproic acid–induced autism-like behaviors accompanied by reduced oxidative stress and neuroinflammation

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by impaired social interaction and repetitive behaviors, with increasing evidence implicating immune dysregulation, oxidative stress, and altered transcriptional regulation. This study investigated the pharmacological effects of Rannasangpei (RNSP), a classical Tibetan formulation, and its active constituent crocin-1 in a valproic acid (VPA)-induced mouse model of ASD. Behavioral tests demonstrated that RNSP and crocin-1 markedly reduced VPA-induced repetitive behaviors and improved social interaction. Biochemical analyses revealed that these effects were associated with restoration of redox balance, characterized by increased activities of catalase and superoxide dismutase and decreased levels of malondialdehyde, together with improvement of the GSH/GSSG ratio. Both treatments also attenuated neuroinflammation, as indicated by reduced expression of IL-6, IL-17A, TNF-α, and Lepr, together with recovery of IL-10 in the prefrontal cortex and cerebellum. Transcriptomic profiling further revealed that RNSP and crocin-1 were associated with widespread transcriptional reorganization in the prefrontal cortex (PFC) and cerebellum. In the PFC, these changes involved pathways related to cell-cell adhesion, morphogenesis, and MAPK signaling. In the cerebellum, the dominant enrichment pattern centered on membrane trafficking and cytoskeleton-related processes, with additional enrichment of oxidative stress response pathways. Integration of pathway analysis and representative gene changes indicated region-specific but functionally related alterations in programs linked to intracellular transport, neuronal structure, and synaptic function. Safety evaluation further showed that neither RNSP nor crocin-1 produced obvious systemic toxicity under the present experimental conditions, as indicated by no consistent treatment-related abnormalities in serum biochemical parameters and histological features of major peripheral organs. These findings indicate that RNSP and crocin-1 were associated with changes in oxidative stress, neuroinflammation, and transcriptional regulation across multiple brain regions, supporting their potential therapeutic relevance in ASD.