Protective Effect of Tongxinluo Capsule on Cerebral Ischemia-reperfusion Injury and Its Mechanism of Action / 中国实验方剂学杂志

ABSTRACT

Objective:To study the protective effect of Tongxinluo capsule on cerebral ischemia-reperfusion injury, and explore the mechanism of Tongxinluo capsule in treating cerebral ischemia through network pharmacology. Method:The C57BL/6 mouse middle cerebral artery occlusion(MCAO) model was established by improved suture method and divided into sham operation group, model group, low, medium and high-dose Tongxinluo groups (crude drug 1,2,4 g·kg-1, intragastric administration), Aspirin group (2.055 g·L-1, intraperitoneal injection). Then, neurological function score and 2,3,5-triphenyltetrazole chloride(TTC) staining method were used to determine the infarct size of mice at 24, 48, 72 h by cerebral ischemia-reperfusion. First, chemical constituents of Tongxinluo capsule were screened from the BATMAN-TCM database, and the targets were analyzed. Then, gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were performed, and traditional Chinese medicine(TCM)-active ingredient-target network was constructed. Finally, the multi-dimensional pharmacological mechanism of Tongxinluo capsule in the treatment of cerebral ischemia was predicted. Result:Longa score, HE staining and TTC staining all suggested that Tongxinluo capsule could alleviate brain injury in mice after cerebral ischemia and reperfusion, and the improvement degree of Tongxinluo capsule on brain injury was gradually enhanced with the increase of Tongxinluo capsule dose. A total of 132 active components and 240 intersection targets, including cyclic adenosine monophosphate(cAMP)-dependent protein kinase catalytic subunit alpha (PRKACA), adenylate cyclase 1(ADCY1), serine/threonine kinase 1 (Akt1), dopamine receptor D2 (DRD2) and discs large homolog 4 (DLG4) were screened from 12 TCM in Tongxinluo capsule. GO was enriched in cationic channel activity, ion gated channel activity, gate channel activity, neurotransmitter receptor activity, ion channel activity, etc. KEGG was enriched in cAMP signaling pathway, calcium signaling pathway, neuroactive ligand-receptor interaction, cyclic guanosine monophosphate(cGMP)/protein kinase G(PKG) signaling pathway and dopaminergic synaptic signaling pathway. Conclusion:Tongxinluo capsule can alleviate brain damage in mice with cerebral ischemia-reperfusion, and achieve brain protection through multiple targets and multiple links. Network pharmacology reveals effective components，targets and pathway of Tongxinluo capsule in the treatment of cerebral ischemia, which provides theoretical support for the mechanism of Tongxinluo capsule in the treatment of cerebral ischemia.