Santalum album L. alleviates cardiac function injury in heart failure by synergistically inhibiting inflammation, oxidative stress and apoptosis through multiple components

Abstract

Background: Heart failure (HF) is a complex cardiovascular syndrome with high mortality. Santalum album L. (SAL) is a traditional Chinese medicine broadly applied for various diseases treatment including HF. However, the potential active compounds and molecular mechanisms of SAL in HF treatment are not well understood.

Methods: The active compounds and possible mechanisms of action of SAL were analyzed and validated by a systems pharmacology framework and an ISO-induced mouse HF model.

Results: We initially confirmed that SAL alleviates heart damage in ISO-induced HF model. A total of 17 potentially active components in SAL were identified, with Luteolin (Lut) and Syringaldehyde (SYD) in SAL been identified as the most effective combination through probabilistic ensemble aggregation (PEA) analysis. These compounds, individually and in their combination (COMB), showed significant therapeutic effects on HF by targeting multiple pathways involved in anti-oxidation, anti-inflammation, and anti-apoptosis. The active ingredients in SAL effectively suppressed inflammatory mediators and pro-apoptotic proteins while enhancing the expression of anti-apoptotic factors and antioxidant markers. Furthermore, the synergistic effects of SAL on YAP and PI3K-AKT signaling pathways were further elucidated.

Conclusions: Mechanistically, the anti-HF effect of SAL is responsible for the synergistic effect of anti-inflammation, antioxidation and anti-apoptosis, delineating a multi-targeted therapeutic strategy for HF.

Keywords: Santalum album L.; Heart failure; Synergistic effect; Systems pharmacology.

Fig. 1

Workflow for SAL alleviates cardiac function injury in heart failure by synergistically inhibiting inflammation, oxidative stress and apoptosis through multiple components

Fig. 2

The effect of SAL on myocardial injury in HF (n = 6). A The in vivo experimental schema. B The appearance of heart. C HE staining. Scale bar, 50 μm. Yellow arrow: infiltration of inflammatory cells. Green arrow: rupture and disintegration of the heart muscle fibres. Black arrow: interstitial space of the myocardium. D HW/BW (Heart weight/body weight ratio in mice). E Lung wet /dry weight. F and G BNP, ANP, cTNT and LDH levels in serum and heart tissue. BNP, ANP, cTNT, pg/ml. LDH, ng/ml. * a significant difference between ISO group and control group (P < 0.05); # displayed significant difference vs ISO group. ISO isoproterenol, SAL Santalum album L., ASA aspirin

Fig. 3

The Compound-target (C-T) network analysis. A The C-T network. B The degree of the active ingredients in SAL. C Percentage of active ingredient degree value. The red nodes correspond to targets, and the blue nodes to SAL’s active compounds. M1, luteolin; M16, syringaldehyde; M6, oleic acid

Fig. 4

Therapeutic effect of active ingredients of SAL on the ISO-induced mouse model of HF. A ECG parameters assessment for various groups. B Heart rate, BPM: beats per minute. C Cardiac Anp relative mRNA level. D and E Serum levels of BNP and Tn-T measured in mice subjected to three different groups. *P < 0.05 compared to ISO group, #P < 0.05 compared to COMB group

Fig. 5

Representative histological results of the heart tissue staining. A H&E, Masson trichrome and wheat germ agglutinin (WGA) stains. Scale bar 50 μm. H&E staining, blue arrow: interstitial space of the myocardium. B HW/BW. C Quantification of the areas of cardiac fibrosis on sections stained with Masson's trichrome. D The cross-sectional area of cardiomyocytes (fold change) calculated from sections stained with WGA-FITC. #P < 0.05 in comparison with the COMB group, *P < 0.05 in comparison with the ISO group

Fig. 6

Biological mechanism analysis of SAL. A GOBP analysis. B the Target-Pathway (T-P) network. Potential targets and the KEGG pathways are represented by blue and purple nodes, respectively. And edges between purple and blue nodes suggest the destination on that pathway. C The representative HF pathway and therapeutic modules of SAL. Purple rectangles represent the targets of active ingredients in SAL, red rectangles represent the targets of HF pathway, respectively

Fig. 7

Functional enrichment experiment verification. A and B IL-2, IL-6, TNF-α, Cleaved-caspase3, caspase3, Bcl2 and BAX protein levels. C mRNA levels of Sod and Cat were measured in vivo. D The ratio of caspase3 to Cleaved caspase3. E Bcl-2/BAX ratio. Significant differences between groups marked with different letters. #P < 0.05 in comparison with the COMB group, *P < 0.05 in comparison with the ISO group

Fig. 8

SAL’s effect on YAP and PI3K/AKT signaling pathway. A Detection and quantification of the expression of PI3K, p-PI3K, AKT, p-AKT, YAP and p-YAP proteins. YAP activation was detected by western blotting by observing the de-phosphorylation of YAP. B mRNA levels of PI3KCG in ventricular tissue of different groups. Groups marked with different letters were significantly different. The symbol # indicates a P value of less than 0.05 when compared with the COMB group, while the symbol * indicates a P value of less than 0.05 when compared with the ISO group