Network pharmacology approach and experimental verification of Dan-Shen Decoction in the treatment of ischemic heart disease

Abstract

Context: Dan-Shen Decoction, which is composed of Danshen, Tanxiang and Sharen, has a good therapeutic effect on ischemic heart disease (IHD). However, systematic research on the exact mechanism of action of Dan-Shen Decoction is still lacking. The anti-IHD effect of Dan-Shen Decoction was examined in this study using a systematic pharmacological method.

Objective: This study validates the efficacy and explores the potential mechanisms of Dan-Shen Decoction in treating IHD by integrating network pharmacology analyses and experimental verification.

Materials and methods: The active components, critical targets and potential mechanisms of Dan-Shen Decoction against IHD were predicted by network pharmacology and molecule docking. H9c2 cells were pretreated with various 1 µg/mL Dan-Shen Decoction for 2 h before induction with 1000 µmol/L CoCl₂ for 24 h. The cell viability was detected by CCK8, and protein expression was detected by western blots.

Results: The network pharmacology approach successfully identified 69 active components in Dan-Shen Decoction, and 122 potential targets involved in the treatment of IHD. The in vitro experiments indicate that the anti-IHD effect of Dan-Shen Decoction may be closely associated with targets such as AKT1 and MAPK1, as well as biological processes such as cell proliferation, inflammatory response, and metabolism.

Conclusions: This study not only provides new insights into the mechanism of Dan-Shen Decoction against IHD, but also provides important information and new research ideas for the discovery of anti-IHD compounds from traditional Chinese medicine.

Keywords: H9c2 cells; Molecular docking; mechanism.

Figure 1.

Venn diagram of the targets of Dan-Shen Decoction and IHD.

Figure 2.

Relationship network between potential therapeutic targets against IHD and active components of Dan-Shen Decoction (DS: Danshen; TX: Tanxiang; SR: Sharen).

Figure 3.

PPI network of potential targets of Dan-Shen Decoction against IHD (inner circle is the top 15 hub proteins in degree value).

Figure 4.

Key functional modules of Dan-Shen Decoction against IHD.

Figure 5.

Results of GO enrichment and KEGG pathway analyses of Dan-Shen Decoction targets (A. Molecular function, Biological process, Cellular component; B. KEGG pathway).

Figure 6.

Docking diagram of main active compounds and potential targets (A. Molecular docking with AKT1, B. Molecular docking with MAPK1).

Figure 7.

Hypoxia induced by CoCl₂ in H9c2 cardiomyocytes. A–C. Cell viability of H9c2 cells stimulated with different concentrations of CoCl₂ (400, 600, 800 and 1000 µmol/L) for 12, 24 and 36 h. *p < 0.05, **p < 0.01 compared with the 0 µmol/L CoCl₂ group. n = 3. D. Cell viability of H9c2 cells induced with various concentrations of Dan-Shen Decoction (1, 10, 50 and 100 µg/mL) for 24 h. E. Cell viability of H9c2 cells pretreated with various concentrations of Dan-Shen Decoction (1, 10, 50 and 100 µg/mL) for 2 h before induction with 1000 µmol/L CoCl₂ for 24 h. **p < 0.01 compared with the control group. #p < 0.05, compared with the CoCl₂-treated and untreated Dan-Shen Decoction groups (n = 3).

Figure 8.

Western blot analyses showed the effect of Dan-Shen Decoction (1 μg/mL) on the expression levels of PI3K/AKT and MAPK signalling pathway-related proteins in control and CoCl₂-induced hypoxic H9c2 cells. The results are shown as the mean ± SEM. **p < 0.01 compared with the control group, #p < 0.05, compared with the CoCl₂-treated group (n = 3).