Akt-1 x MMP9 x TNF x albumin x caspase 3

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder characterized by chronic inflammation, oxidative stress, and amyloid beta (Aβ) aggregation. Prolonged exposure to heavy metals such as aluminium (Al) promotes reactive oxygen species (ROS) generation and neuroinflammation, contributing to Aβ aggregation, neuronal damage, and cognitive decline. Although conventional drugs show therapeutic potential, their use is often limited by adverse side effects. Phytochemicals with antioxidant, anti-inflammatory, and anticholinesterase properties are safer alternatives. This study explored the neuroprotective potential of agnuside (AGN) against aluminium chloride (AlCl₃)-induced AD pathology through network pharmacology and experimental approaches. A total of 108 overlapping targets of AGN and AD were identified, with key nodes including TNF, AKT1, ALB, EGFR, ESR1, CASP3, MMP9, SRC, HSP90AA1, and PPARG. Gene ontology and KEGG pathway analyses suggested that AGN may modulate key molecular pathways implicated in AD, including TNF signaling and the PI3K/AKT signaling. Molecular docking confirmed that AGN has strong binding affinities with TNF, AKT1, ALB, MMP9, and CASP3. SH-SY5Y cells and zebrafish larvae exposed to AlCl₃ exhibited AD pathology, which was notably attenuated by AGN, as indicated by reductions in oxidative stress, apoptosis, and neuroinflammation, confirmed through acridine orange (AO), reactive oxygen species (ROS), and neutral red staining. This protective effect was further validated by enzymatic assays and gene expression analysis, confirming its antioxidant and anti-apoptotic potential. These findings suggest that AGN is a promising phytotherapeutic candidate for counteracting AlCl₃-induced neurotoxicity and could serve as a potential therapeutic agent for AD intervention.

As a well-known traditional Chinese medicine, Amomi fructus (A. fructus) (Sharen) has been used therapeutically to treat gastrointestinal illnesses, including gastric ulcer (GU). The mechanism underlying this impact is still not fully known, though.

To investigate the hidden mechanism by which A. fructus influences the pathogenesis of GU, we employed network pharmacology approaches and in vivo validated studies.

Multiple public databases were used to compile information on bioactive compounds, potential targets of A. fructus, and associated genes of GU. Then, the STRING database's protein-protein interaction (PPI) data of the drug-disease overlapping gene targets was obtained, and the core targets for A. fructus against GU were discovered. Additionally, molecular docking was done to examine the binding capabilities of the active substances and core targets. Then, the pathways of A. fructus that target GU were examined using the Annotation, Visualization and Integrated Discovery (DAVID)'s Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway studies. In a mouse model of acute stomach mucosal damage brought on by absolute ethanol, the findings of network pharmacology were finally validated.

In total, 610 targets derived from the 196 bioactive compounds in A. fructus, were discovered, and along with 115 A. fructus target genes for therapy of GU. Then, ten core targets associated with apoptosis and inflammation were determined based on node degree, and ALB, AKT1, TNF, EGFR, MAPK3, CASP3, MMP9, STAT3, SRC, and HRAS were identified as promising therapeutic targets of A. fructus against GU. The results of molecular docking also revealed that 65 active compounds had strong binding activity with the core targets, with volatile chemicals being the most significant active ingredients. So, for following in vivo tests, A. fructus volatile oils (AVO) were used. The KEGG analysis showed that the phosphoinositide-3-kinase/protein kinase B (PI3K/AKT) signaling pathway may be crucial for the therapeutic mechanism of GU. In experiments that were validated in vivo, AVO considerably decreased the ulcer area and enhanced the histological appearance of the gastric tissues. In addition, compared to the model group, up-regulated the expression of IGF-1, p-PI3K, and p-AKT and down-regulated the protein levels of TNF-α and Caspase 3 in the stomach tissues.

According to preliminary findings from this work, A. fructus may influence inflammatory response and apoptosis via regulating the PI3K/AKT signaling pathway and associated gene targets. Importantly, our research might offer a theoretical foundation for future research into the intricate anti-GU mechanism of A. fructus.

Kaempferol is a common flavonoid aglycone widely found in plants. It exhibits beneficial therapeutic effects in the treatment of arthritis. However, the effects of kaempferol on gouty arthritis (GA) have not been verified. This study aimed to explore the potential mechanisms by which kaempferol regulates GA by network pharmacology and experimental validation. Potential drug targets for GA were identified with a protein-protein interaction network. Then, we performed a KEGG pathway analysis to elucidate the major pathway involved in the kaempferol-mediated treatment of GA. In addition, the molecular docking was performed. A rat model of GA was constructed to verify the results of network pharmacology analysis and investigate the mechanism of kaempferol against GA. The network pharmacology study indicated that there were 275 common targets of kaempferol and GA treatment. Kaempferol exerted therapeutic effects on GA, in part, by regulating the IL-17, AGE-RAGE, p53, TNF, and FoxO signaling pathways. Molecular docking results showed that kaempferol stably docked with the core MMP9, ALB, CASP3, TNF, VEGFA, CCL2, CXCL8, AKT1, JUN, and INS. Experimental validation suggested that kaempferol eased MSU-induced mechanical allodynia, ankle edema, and inflammation. It significantly suppressed the expression of IL-1β, IL-6, TNF-α, and TGF-β1 and restored Th17/Treg imbalance in MSU-induced rats and IL-6-induced PBMCs. Kaempferol also affected RORγt and Foxp3 through IL-17 pathway. The present study clarifies the mechanism of kaempferol against GA and provides evidence to support its clinical use.