Integrating bioinformatics and ferroptosis to reveal the protective mechanism of Astragaloside IV on chronic heart failure rats

Abstract

Ferroptosis is an important pathological mechanism of chronic heart failure (CHF). This study aimed to investigate the protective mechanism of Astragaloside IV (AS-IV) on CHF rats by integrating bioinformatics and ferroptosis. CHF-related targets and ferroptosis-related targets were collected. After the intersection, the common targets were obtained. The PPI network of the common targets was constructed, and topological analysis of the network was carried out. The target with the highest topological parameter values was selected as the key target. The key target p53 was obtained through bioinformatics analysis, and its molecular docking model with AS-IV was obtained, as well as molecular dynamics simulation analysis. The rat models of CHF after myocardial infarction were established by ligation of left coronary artery and treated with AS-IV for 4 weeks. AS-IV treatment significantly improved cardiac function in CHF rats, improved cardiomyocyte morphology and myocardial fibrosis, reduced mitochondrial damage, decreased myocardial MDA and Fe²⁺ content, increased GSH content, inhibited the expression of p53 and p-p53, and up-regulated the expression of SLC7A11 and GPX4. In conclusion, AS-IV improved cardiac function in CHF rats, presumably by regulating p53/SLC7A11/GPX4 signaling pathway and inhibiting myocardial ferroptosis.

Fig. 1

The workflow chart of bioinformatics analysis and animal experiment.

Fig. 2

CHF-related targets and ferroptosis-related targets. (a) The volcano plot of differentially expressed genes. (b) The heat map of partial differentially expressed genes. (c) Venn diagram of CHF-related targets. (d) Venn diagram of ferroptosis-related targets. (e) Venn diagram of CHF-related targets and ferroptosis-related targets.

Fig. 3

PPI network of common targets and network of core targets. (a) PPI network of common targets. (b) First screening of common targets. (c) Second screening of core targets. (d) Screening of key target p53 through topological analysis.

Fig. 4

GO and KEGG enrichment analyses of common targets. (a) The barplot of GO enrichment analysis. (b) The bubble chart of GO enrichment analysis. (c) The barplot of KEGG enrichment analysis. (d) The bubble chart of KEGG enrichment analysis. (e) The map of ferroptosis pathways.

Fig. 5

Molecular docking model of p53 with AS-IV. (a) Molecular docking model without surface of receptor. (b) Molecular docking model showing surface of receptor.

Fig. 6

MD simulation analysis of p53-AS-IV complex. (a) RMSD curve of p53, AS-IV and p53-AS-IV complex. (b) RMSF curve of p53’s amino acid chains. (c) Rg curve of p53. (d) SASA curve of p53. (e) H-bonds of p53-AS-IV complex.

Fig. 7

AS-IV improved cardiac function in CHF rats. (a) Representative echocardiographic images of rats after 4 weeks of AS-IV administration. (b) The result of LVFS at 4 weeks after coronary ligation, Sham (n = 9), Modeling (n = 20). (c–i) The results of LVFS, LVIDs, LVIDd, LVESV, LVEDV, heart weight index and serum NT-proBNP content (n = 9). **P < 0.01 vs. Sham group; ^##P < 0.01, ^#P < 0.05 vs. Model group.

Fig. 8

AS-IV alleviated myocardial histomorphological changes in CHF rats. (a,b) Representative images of HE staining. (c,g) Representative images and quantification of the cardiomyocyte cross-sectional area in the non-infarcted zone (n = 3). (d,h) Representative images of Masson staining and quantification of the left ventricular infarct size (n = 3). (e,i) Representative images and quantification of the fibrosis in the infarcted border zone (surgical area for the Sham group, n = 3). (f,j) Representative images and quantification of the fibrosis in the non-infarcted zone (n = 3). **P < 0.01 vs. Sham group; ^##P < 0.01 vs. Model group.

Fig. 9

AS-IV alleviated myocardial mitochondrial damage in CHF rats. Representative images of TEM.

Fig. 10

AS-IV decreased MDA content and Fe²⁺ accumulation and increased GSH content in CHF rats. (a) The result of MDA content in myocardium (n = 6). (b) The result of Fe²⁺ content in myocardium (n = 6). (c) The result of GSH content in myocardium (n = 6). **P < 0.01, *P < 0.05 vs. Sham group; ^##P < 0.01, ^#P < 0.05 vs. Model group.

Fig. 11

AS-IV regulated the expression of p53/SLC7A11/GPX4 signaling pathway. (a) The mRNA expression of p53/SLC7A11/GPX4 signaling pathway (n = 3). (b) The representative Western Blotting bands of p53/SLC7A11/GPX4 signaling pathway. The original blots/gels were presented in Supplementary Figure. (c) The protein expression of p53/SLC7A11/GPX4 signaling pathway (n = 3). **P < 0.01, *P < 0.05 vs. Sham group; ^##P < 0.01, ^#P < 0.05 vs. Model group.