The Potential Mechanism of Zishen Yutai Pills against Threatened Abortion: An Approach Involving Network Pharmacology and Experimental Evidence

Abstract

Threatened abortion (TA) is the most common complication in early pregnancy and is caused by anxiety and depressive symptoms. The Zishen Yutai Pill (ZYP) is a traditional herbal formula that is commonly used to treat TA. However, the pharmacological mechanisms underlying the effect of ZYP have yet to be elucidated. To disclose the mechanism of ZYP in the treatment of TA, first, we identified the chemical constituents of ZYP from multiple databases and then predicted the potential targets of TA by applying the GeneCards database. A protein-protein interaction (PPI) network was then constructed to allow the screening of hub targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGGs) enrichment analyses were also performed and the Database for Annotation Visualization and Integrated Discovery server was used to identify critical biological processes and signaling pathways. Cytoscape software was used to construct a Compound-Target-Pathway. Furthermore, we analyzed ZYP by UPLC-Q-TOF/MS, then the RU486-induced TA rat model was established, and the reliability of the network pharmacology prediction results was verified. Finally, the mechanisms responsible for the action of ZYP on TA were revealed by qRT-PCR and molecular docking. Database screening identified a total of 161 active compounds in ZYP and 324 TA-related targets. And, we identified 42 compounds from ZYP by UPLC-Q-TOF/MS. Inflammation and apoptosis were identified as the main biological processes. GO and KEGG analyses identified that the MAPK and PI3K/Akt pathways were the key functional pathways that respond to ZYP. The results showed that ZYP treatment significantly increased maternal weight, significantly increased the levels of estradiol and progesterone, and attenuated histopathological changes in a rat model of TA. Data indicated that ZYP treatment improved pregnancy outcomes in the rat model of TA. QRT-PCR data showed that ZYP reduced inflammation and apoptosis by regulating the MAPK and PI3K/Akt pathways. In addition, molecule docking results identified a range of key compounds, including Pik3a, Mapk14, Mapk1, Mapk3, Mapk8 Tnf, Il6, and Cas8. In summary, we performed network pharmacological analysis and experimental validation and identified that ZYP exerts an effect on TA by regulating the MAPK and PI3K/Akt pathways and by inhibiting the expression levels of proinflammatory cytokines and genes related to apoptosis.