Objective:

2

To explore the neuroprotective effects and mechanism of Tanreqing Injection (TRQ) on treating ischemic stroke based on network pharmacology and

in vivo

experimental validation.

Methods:

2

The che

mical compounds of TRQ were retrieved based on published data

with targets retrieved from PubChem

Therapeutic Target Database and DrugBank. Network visualization and analysis were performed using Cytoscape

with protein-protein interaction networks derived from the STRING database. Enrichment analysis was performed using Kyoto Encyclopedia of Genes Genomes pathway and Gene Ontology analysis. In

in vivo

experiments

the middle cerebral artery occlusion (MCAO) model was used. Infarct volume was determined by 2

3

5-triphenyltetrazolium hydrochloride staining and protein expressions were analyzed by Western blot. Molecular docking was performed to predict ligand-receptor interactions.

Results:

2

We screened 81 chemical compounds in TRQ and retrieved their therapeutic targets. Of the targets

116 were therapeutic targets for stroke. The enrichment analysis showed that the apelin signaling pathway was a key pathway for ischemic stroke. Furthermore

in

in vivo

experiment we found that administering with intraperitoneal injection of 2.5 mL/kg TRQ every 6 h could significantly reduce the infarct volume of MCAO rats (

P

<

0.05). In addition

protein levels of the apelin receptor (APJ)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway were increased by TRQ (

P

<

0.05). In addition

41 chemical compounds in TRQ could bind to APJ.

Conclusions:

2

The neuroprotective effect of TRQ may be related to the APJ/PI3K/AKT signaling pathway. However

further studies are needed to confirm the findings.