## 2 Results and Discussion
### Network Pharmacology Analysis
Figure 1: Network pharmacology analysis for prediction of the underlying mechanism of spilanthol in the attenuation of neurodegenerative diseases. (**A**) Chemical structure of spilanthol, MW = 221.339 g/mol. (**B**) Intersection between spilanthol target genes and differentially expressed genes in neurodegenerative diseases. (**C**) Protein–protein interactions of 90 common targets. (**D**) Network of the top 10 hub genes using Maximal Clique Centrality analysis. Red to yellow colors signify varying degrees of scores, with red typically indicating a higher score and yellow indicating a lower score. SP, spilanthol; NDDs, Neurodegenerative diseases. Suppression of the TLR4 pathway has been proven to produce favorable outcomes against many NDDs, including AD, PD, ALS, MS, and HD [34,35]. Interestingly, suppression of TLR4 signaling in AD has been proven to improve cognitive dysfunction in streptozotocin-induced murine models of AD [36]. Similarly, inhibition of TLR4-induced microglial polarization through the administration of TLR4-specific inhibitor TAK-242 has significantly improved neurological function, reduced the level of M1 microglia, and inhibited neuronal apoptosis [37]. Although, TLR4 signaling produces a protective effect in acute phases of PD, by inducing the clearance of \(\alpha\)-synuclein, chronic activation of the TLR4 signaling pathway was found to exacerbate neurodegeneration by inducing chronic neuroinflammation, and suppression of TLR4 signaling was found to alleviate the features of \(\alpha\)-synucleinopathies, excessive inflammation, and progress of neurodegeneration [38]. Importantly, compounds capable of suppressing TLR4/NF-\(\kappa\)B signaling have significantly reduced the loss of dopaminergic neurons and have improved locomotor impairments in LPS-induced murine models of Parkinson's disease [39,40]. As per the findings of previous studies, suppression of TLR signaling has produced therapeutic benefits in NDDs and, interestingly, splianthol has interacted with multiple proteins in the TLR pathway in favor of attenuating neurodegenerative diseases. Figure 2: Gene ontology (GO) and KEGG enrichment analysis of spianthol- and neurodegenerative disease-related targets. Illustrations of enriched biological processes (**A**), cellular components (**B**), molecular functions (**C**), and KEGG (**D**) enrichment pathway analyses. The color of each GO and KEGG term indicates the \(p\)-value, and the area of the circle indicates the number of enriched genes. Considering the AD pathway, splianthol demonstrated substantial potential to modulate with multiple pathophysiological pathways by interacting with 27 target proteins involved in AD pathophysiology, including NMDAR, CASP8, BACE, JNK, and mTOR, in addition to targets in the inflammatory pathway (Supplementary Figure S2) [41,42]. Considering the complex pathophysiology and potential of spilanthol to interact with multiple targets involved in different pathological pathways, future studies are warranted to comprehensively evaluate the potential of spilanthol in attenuating AD focusing on different pathways by evaluating different biomarkers [43,44].