Network Pharmacology and Experimental Validation-based Approach to Explore the Effect and Mechanism of Scutellaria baicalensis-Rhodiola rosea Herbal Pair against Ischemic Stroke

Objective: An integrated approach combining network pharmacology and experimental verification was used to elucidate the effects and mechanisms of the Scutellaria baicalensis (Huangqin [HQ]) and Rhodiola rosea (Hongjingtian [HJT]) herbal pair in treating ischemic stroke (IS). Materials and Methods: A compound–disease network for the HQ-HJT herbal pair and IS was constructed for the network pharmacology analysis. In addition, the efficacy and targets of the HQ-HJT herbal pair were verified using a classical in vitro IS model and molecular docking methods. Results: A total of 82 overlapping targets were identified between the HQ-HJT herbal pair and IS. Kaempferol was the most active compound in the compound–disease network. The protein-protein interaction network analysis showed that Interleukin-6, tumor necrosis factor-α (TNF-α), and HIF-1α were core targets. Kyoto Encyclopedia of Genes and Genomes analysis identified that the HQ-HJT herbal pair exerts therapeutic effects on IS through the TNF signaling and other related pathways. Our experimental analyses confirmed that the key components (kaempferol, salidroside, and baicalein) had neuroprotective effects in the oxygen-glucose deprivation/reoxygenation injury model. Notably, kaempferol suppressed inflammatory responses and showed strong binding affinity to six key targets within the HIF-1α/NLRP3/Caspase-1 signaling axis. Conclusions: HQ-HJT may exert an anti-IS effect by inhibiting the inflammatory cascade. Kaempferol may serve as a key bioactive compound in the HQ-HJT pair.