Integrating Network Pharmacology and an Experimental Model to Investigate
the Effect of Zhenwu Decoction on Doxorubicin-Induced Heart Failure
Authors: Hu Yiran , Qu Huiyan , Zhou Hua 
Published: April 2023

Background:

Doxorubicin-induced heart failure is a clinical problem that needs to be
solved urgently. Previous studies have confirmed that Zhenwu Decoction, a traditional Chinese
medicine compound, can effectively improve chronic heart failure. However, its interventional effect
on Doxorubicin-induced heart failure has not yet been investigated. In this study, we investigated
the therapeutic effect and potential mechanism of Zhenwu Decoction on Doxorubicininduced
heart failure through animal experiments and network pharmacology.

Objective:

The study aimed to investigate the therapeutic effect and potential mechanism of Zhenwu
Decoction (ZWD) on Doxorubicin-induced heart failure.

Methods:

A heart-failure mouse model was established in 8-week-old male C57/BL6J mice using
Doxorubicin, and the mice were then treated with ZWD for a 4-week period. Firstly, network
pharmacology was conducted to explore the potential active components and molecular mechanisms
of ZWD on Doxorubicin-induced heart failure. Next, we conducted an in vivo study on the
effect of ZWD on Doxorubicin-induced heart failure. After the intervention, the cardiac function
and levels of cardiac function injury marker in serum were measured to evaluate the therapeutic effect
of ZWD on cardiac function. Then HE staining and Masson staining were used to evaluate the effect
of ZWD on myocardial pathology, and biochemical method was used to detect the effect of ZWD on
total antioxidant capacity and inflammation, and finally, Western blot was used to detect TGFβ,
Smad-3, and collagen I protein expression levels to evaluate its effect on myocardial fibrosis.

Results:

In Doxorubicin-induced heart failure mice, ZWD improved cardiac function and reduced
the levels of CK-MB, NT-proBNP, and BNP in the serum, improved myocardial pathology, and
reduced TGFβ, Smad-3 and collagen I protein expression levels to improve myocardial fibrosis.
Network pharmacological analysis showed that ZWD has 146 active ingredients and 248 candidate
targets. Moreover, 2,809 genes were found to be related to Doxorubicin-induced heart failure, and
after screening, 74 common targets were obtained, mainly including IL-6, AKT1, caspase-3,
PPARG, PTGS2, JUN, HSP90AA1, and ESR1. KEGG analysis confirmed that PI3K/AKT and IL-
6/NF-κB signaling pathways were the two main pathways underlying the cardioprotective effects
of ZWD. Finally, in vivo experiments showed that ZWD improved the total antioxidant capacity,
reduced the SOD level, increased the protein expression of PI3K, Akt, Bcl-2, Bax, and caspase-3,
reduced the levels of TNF-α, IL-6, and IL-1β, and decreased the NF-κB p65, IL-6, and TNF-α protein
expression levels.

Conclusion:

In Doxorubicin-induced heart-failure mice, Zhenwu Decoction improved the cardiac
function and myocardial pathology, and improved myocardial fibrosis through the TGFβ/Smad-3
signaling pathway. According to the prediction of network pharmacology, in vivo experiments
demonstrated that Zhenwu Decoction can improve the oxidative stress response, improve myocardial
cell apoptosis through the PI3K/AKT signaling pathway, and improve myocardial inflammation
by reducing the levels of inflammatory factors and by reducing the protein expression of NF-
κB p65, IL-6, and TNF-α.