Salvia miltiorrhiza Bge. processed with porcine cardiac blood alleviates cerebral ischemia reperfusion injury through activating PPARγ/NF-κB pathway via suppressing arachidonic acid metabolism

Ethnopharmacological relevance: Salvia miltiorrhiza Bge. (DS) processed with porcine cardiac blood (PCB-DS) is primarily used to treat cerebral ischemia, in accordance with the traditional principles of the Menghe Medical School. Studies demonstrated that PCB-DS was superior to DS in ameliorating cerebral ischemia-reperfusion injury (CIRI). This effect was related to the modulation of brain inflammation. However, the underlying mechanism and material basis responsible for PCB-DS's superior efficacy over DS in improving CIRI require further investigation.

Aim of the study: To investigate the mechanism and material basis of PCB-DS superiority over DS in ameliorating CIRI.

Materials and methods: The pharmacological activities of PCB-DS and DS were compared by the middle cerebral artery occlusion-reperfusion (MCAO/R) model. Proteomics and metabolomics were used to explore the potential mechanisms underlying PCB-DS's greater efficacy than DS in reducing CIRI. Based on the results, the PPARγ signaling pathway and arachidonic acid (AA) metabolism pathway were analyzed. Subsequently, the brain-penetrating components were analyzed using UPLC-Q-TOF/MS. Finally, molecular docking was performed between proteins related to the PPARγ signaling pathway and active components to confirm the material basis for PCB-DS's superior efficacy over DS.

Results: Compared to DS, PCB-DS better alleviated CIRI by improving neurobehavioral scores, lowering infarct volume, ameliorating brain histopathology, and decreasing oxidative damage indicators. Proteomic analysis revealed that the mechanism underlying PCB-DS and DS was also strongly associated with the regulation of the PPAR signaling pathway. WB results further indicated that PCB-DS more effectively reversed the expression levels of NLRP3, p-NF-κB, and inflammation-related concentrations vs. DS group (p < 0.05). The metabolomics analysis revealed that the impact of PCB-DS and DS was closely related to AA metabolism. PCB-DS better improved the level of AA and 15d-PGJ2 in rat brain vs. DS group (p < 0.05). The brain-penetrating components demonstrated that PCB-DS significantly elevated the concentrations of salvianolic acid B and 9″-methyl salvianolate B vs. DS (p < 0.05). Molecular docking further validated that these two components bind strongly to key targets such as PPARγ, NLRP3, p-IKBα, and p-NF-κB.

Conclusion: The superior effect of PCB-DS over DS in CIRI was to protect brain from inflammation through activating PPARγ/NF-κB pathway via suppressing AA metabolism. The elevated levels of 9″-methyl salvianolate B and salvianolic acid B may contribute to this enhanced effectivity.

Cerebral ischemia-reperfusion injury; Inflammation; Metabolomics; Proteomics; Salvia miltiorrhiza Bge. processed with porcine cardiac blood.