Astragaloside IV promotes pharmacological effect of Descurainia sophia seeds on isoproterenol-induced cardiomyopathy in rats by synergistically modulating the myosin motor

Figures

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  FIGURE 1. Workflow of quantitative proteomics analysis. The heart tissues of rats from seven groups are designated as the control group (CM), ISO-iCM model group (MG), and treated groups with Descurainia sophia seed decoction alone (DS), and combined with Astragalus mongholicus (DSAM), astragaloside IV (DSAS4), astragalus calycosin-7-glucoside (DSC7G), or astragalus polysaccharides (DSAPS), respectively.

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  FIGURE 2. Chemical structure of (A) quercetin-7-O-β-D-glucopyranosyl (1→6)-β-D-glucopyranoside, (B) quercetin, and (C) astragaloside IV.

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  FIGURE 3. Phenotypic and pathological characterization (A–F), left ventricular ejection fraction (LVEF, (A), left ventricular fraction shortening (LVFS, (B), heart index (HWI, (C), left ventricular mass index (LVWI, (D), collagen volume fraction (CVF, (E) of myocardial cells, and plasma CK levels (F) of rats of the control (CG), model (MG), and treated groups with Descurainia sophia seed decoction alone (DS), and Descurainia sophia seeds plus Astragalus mongholicus decoction (DSAM), Descurainia sophia seeds decoction plus astragaloside IV (DSAS4), calycosin-7-glucoside (DSC7G), or Astragalus polysaccharides (DSAPS). The results of LVEF, LVFS, HWI, LVWI, CVF, and CK were measured by randomly selecting six rats (n = 6) from each group and are represented as mean ± SD. *p < 0.05 and **p < 0.01, ***p < 0.001 and ****p < 0.0001, represent statistical significance and high significance, respectively. (G–M) H&E (400×) and (N–T) Masson’s (200×) trichrome staining of heart tissues of all groups. For Masson’s staining, collagen fibers were stained in blue, and myocardial cell in dark red.

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  FIGURE 4. Differential expression proteomics of the heart of rats in the control group (CG) and ISO-iCM model group (MG). (A) Number of proteins identified in three parallel replicates. (B) Volcanic map of the proteins identified in both the control (CG) and model (MG) groups with various abundance ratios (ARs) and p-values. Orange point refers to a protein with a log2AR (MG vs. CG) of ≥0.59 or ≤ −0.59 and a p-value of <0.05; blue to a protein with a log2AR of > −0.59 or <0.59 and a p-value of <0.05, gray to a protein with a p-value of >0.05. (C) Fold changes (FCs) of 25 differentially expressed proteins in the heart of the model group (MG) compared to those in the control group (CG). The insert is the Western blotting bands of the representatives of DEPs, NPPA, TMP1, and TMP2 in the heart of rats in CG, MG, and the treated group with Descurainia sophia seeds decoction plus astragaloside IV (DSAS4). GAPDH was used as an internal reference.

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  FIGURE 5. Differentially expressed proteins (DEPs) were detected in the heart of rats with ISO-iCM before and after treatments, compared to the controls. (A) Abundance ratios (model or treated groups vs. control group) of 25 DEPs in the model group (MG), treated group with Descurainia sophia seed decoction alone (DS), and Descurainia sophia seeds plus Astragalus mongholicus decoction (DSAM), Descurainia sophia seed decoction plus astragaloside IV (DSAS4), calycosin-7-glucoside (DSC7G), or astragalus polysaccharides (DSAPS). (B) Principal component analysis (PCA) is based on 213 proteins identified in all groups with a p-value <0.05. (C,D) Loading scores discriminate the model (MG) and treated groups, DS, DSAM, DSAS4, DSC7G, and DSAPS, over the control group (CG).

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  FIGURE 6. Bioinformatics analysis of differentially expressed proteins (DEPs) in the hearts of rats with ISO-iCM compared to the controls (A) Protein–protein interaction network of the 25 DEPs. (B) KEGG pathways and biological processes in which the 25 DEPs are associated with.

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  FIGURE 7. Diagrammatic process of isoproterenol-induced heart dysfunction in rats. It shows that isoproterenol binds to ADRB2 and upregulates the expression of NPPA, MYH6, and MYH7 via activating the transcriptional regulators STAT3 and GATA4. The inset shows the abundance ratio of the selected proteins expressed in the heart of controls (CG) and models (MG).

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  FIGURE 8. NPPA-triggered signaling pathway works together with the calcium signaling pathway to modulate contractility and remodeling of the myocardium.