Inhibiting Effect and Mechanism of Aconitum tanguticum (Maxim.) Stapf on Intestinal Fibrosis of CCD-18Co Cells

Abstract

The objective of the study was to explore the potential signaling mechanism of Aconitum tanguticum (Maxim.) Stapf (ATS) and its impact on intestinal fibrosis in vitro. Network pharmacology was used to screen the active components of ATS and predict their potential targets in intestinal fibrosis. The protein–protein interaction network graph was constructed using drug–disease intersection targets retrieved from the Search Tool for Retrieval of Interacting Genes/Proteins database. The network diagram was analyzed using Cytoscape 3.6.1’stopology function. The gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted using the database for annotation, visualization, and integrated discovery platform. Intestinal fibroblast model in vitro was constructed using transforming growth factor-β1 (TGF-β1)-induced CCD-18Co cells. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to verify the network pharmacology-predicted antifibrotic signaling pathway of ATS and the traditional antifibrotic signaling pathway. Network pharmacology revealed that there were 19 active components in ATS, suggesting that ATS could be involved in the regulation of mitogen-activated protein kinase 1 (MAPK1), PIK3CA, MAPK3, and other important targets that are present in cancer pathways, including proteoglycans, influenza A, and phosphatidylinositol-3-kinase (PI3K)-protein kinase B (Akt) signaling pathways. The results of quantitative reverse transcription-polymerase chain reaction (qRT-PCR) demonstrated that 0.1–1 μg/mL ATS may suppress tissue inhibitor of metalloproteinase 1, collagen I, and alpha-smooth muscle actin expression levels. The expression of the TGF-β/suppressor of the mother against decapentaplegic (Smad) and PI3K/Akt signaling pathways was controlled by ATS. In addition, ATS inhibited the PI3K/Akt pathway by reducing the expression of MAPK1, HSP90AA1, and PIK3CA. It has been confirmed that ATS is a multipathway and multitarget treatment for intestinal fibrosis. This study suggested that ATS might alleviate intestinal fibrosis by blocking both the TGF-β/Smad and the PI3K/Akt pathway.