Pharmacological Research - Modern Chinese Medicine (Jun 2023)
A mechanistic study on the Wenxin formula for atherosclerosis treatment based on miRNA sequencing and network pharmacology
Abstract
Context: The Wenxin formula, developed by Professor Hongxin Cao, has achieved remarkable efficacy in clinical CAD treatment. Objectives: 1. Evaluate the effectiveness of the Warm Heart formula in treating atherosclerosis; 2. Investigate the mechanism of action of the Wenxin formula in treating atherosclerosis. Methods: 1. HE and oil red staining evaluation of the modeling and efficacy of the Wenxin formula on atherosclerotic lesions; 2. On the arterial tissues of atherosclerotic APOE-/- mice, high-throughput miRNA sequencing was performed to identify key miRNAs effectively regulated by the Wenxin formula, and the results were entered into the Targetscan and Miranda databases for target gene prediction and KEGG enrichment analysis; 3. The active ingredient information of the Wenxin formulation was obtained by searching the TCMSP database, while the target information was obtained by searching the Uniprot website. The integrated data were imported into the Cytoscape 3.5.1 software to generate a ''Wenxin formula-compound-target'' visual network diagram. The DisGeNET, Genecard, DrugBank, OMIM, GAD and TTD databases were searched for atherosclerosis targets, the intersection of disease and Wenxin formula targets were entered into the String website and Cytoscape 3.5.1 to build a PPI network, and the Metascape platform was utilized to perform GO and KEGG enrichment analysis; 4. The proteins on key PI3K-Akt pathways (IGF1R, ERK2, PIKCA, and PRKCA) were validated using Western blotting. Results: 1. HE and oil red staining demonstrated that the Wenxin formula could significantly improve lesions associated with AS; 2. High-throughput sequencing was used to screen 30 ''lesion'' miRNA genes, and the Wenxin formula can effectively regulate 18 of them; 3. Wenxin formula was able to regulate 196 of the 3,127 disease-target genes that were screened. Beta-sitosterol, stigmasterol, and naringenin may play crucial roles in the Wenxin formula, and the PI3K-Akt pathway may be a crucial component of the Wenxin formula; 4. Western blot experiments demonstrated that the Wenxin formula significantly impacts the PI3K-Akt pathway of IGF1R, ERK2, PIKCA, and PRKCA. Conclusion: 1. Wenxin formula can improve arterial lesions in mice with AS; 2. Wenxin formula can enhance miRNA expression in some AS ''lesions''; 3. Beta-sitosterol, stigmasterol, naringenin, and other compounds in the Wenxin formula may play a crucial role, and the PI3K-Akt pathway may be the key determinant.
