Mechanism of Dahuang Mudan Decotion in the treatment of colorectal cancer based on network pharmacology and experimental validation

Abstract

Objective: The objective of this study was to assess the pharmacological activity and therapeutic mechanism of Dahuang Mudan Decotion (DHMDD) for colorectal cancer using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS), network pharmacology and in vitro experiments.

Methods: The chemical components of DHMDD were identified by UPLC-MS. Network pharmacological analysis was utilized to screen the active ingredients and targets associated with DHMDD for colorectal cancer. Based on the results of network pharmacology, the potential mechanism of DHMDD on colorectal cancer predicted was experimentally studied and verified in vitro.

Results: DHMDD primarily exerts its effects on colorectal cancer through 52 active ingredients. AKT1, ESR1, HSP90AA1, JUN, PIK3CA, PIK3CB, PIK3R1, SRC, STAT3, TP53 were the top 10 targets. The top 10 ingredient nodes were Quercetin, Physcione, Pontigenin, Crysophanol, Linolenic acid, Piceatannol, Adenosine, Emodin, Sambunigrin, and Prunasin. The main compounds and the target proteins exhibited strong binding ability in molecular docking studies. The results of cell experiments demonstrated that DHMDD can inhibit the proliferation, invasion and migration of CRC cells through the PI3K/Akt pathway.

Conclusion: Through network pharmacology analysis and cell experiments, this study suggests that DHMDD can exert its therapeutic effects on colorectal cancer through a combination of multiple components and targets.

Keywords: Colorectal cancer; Dahuang Mudan Decotion; Molecular docking; Network pharmacology; Traditional Chinese medicine.

Fig. 1

Representative base peak intensity (BPI) chromatograms of DHMDD in (A) positive and (B) negative modes by UPLC-ESI-Q-TOF/MS. The numbering of identified compounds (1−81) is the same as in Supplementary Table S1.

Fig. 2

(A)the Venn Diagram between constituents-related targets and CRC-related targets; (B)The network of 53 constituents and overlapped CRC-related targets.

Fig. 3

(A) Protein-Protein Interaction Networks of potential targets; (B) topological analysis of potential targets; The color from light to dark indicates the Degree value from small to large; (C) the Interaction Networks of ten core targets.

Fig. 4

Enrichment analysis of potential targets from main active ingredients of DDMDD: (A) Gene ontology terms for biological process; (B) Gene ontology terms for molecular function; (C) Gene ontology terms for cellular component; (D) KEGG pathways.

Fig. 5

(A) Molecular docking heat map; (B)Schematic diagram of molecular docking.

Fig. 6

(A) The graph of DHMDD inhibiting HCT116 growth. (B) Effect of DHMDD on CRC cell clonal formation (**p＜0.01). (C) Effect of DHMDD on CRC cell invasion (**p＜0.01).(D) Effect of DHMDD on CRC cell migration (**p＜0.01).(E) Effect of DHMDD on PI3K, AKT1, PIK3CA and PIK3R1 mRNA expression.(F) Effect of DHMDD on AKT1, p-AKT1, Bcl 2 and Bax protein expression (*p < 0.05, **p < 0.01).