Salvianolic acid D: A potent molecule that protects against heart failure induced by hypertension via Ras signalling pathway and PI3K/Akt signalling pathway

Figures

• 

  Fig. 1. The effects of Sal D treatment on blood pressure and cardiac function after administration for 12 weeks. (A) SBP; (B) DBP; (C) EF; (D) FS; (E) Represented image of ultrasound cardiogram. n = 8. ###, P < 0.001 vs. WKY. ##, P < 0.01 vs. WKY. #, P ​< ​0.05 vs. WKY. ∗∗∗, P ​< ​0.001 vs. SHR. ∗∗, P ​< ​0.01 vs. SHR. ∗, P < 0.05 vs. SHR.

• 

  Fig. 2. The effects of Sal D treatment on myocardial injury. (A) Masson staining images under 200 × magnification and the areas of collagen deposition were indicated by the black arrow; (B) CVF; (C–D) Quantification of expression levels of ANP and BNP mRNA in heart tissue were measured using Real-time PCR (n = 3). 2−△△Ct method was used to analyze relative gene expression levels. (∗P ​< ​0.05, ∗∗P ​< ​0.01 and ∗∗∗P < 0.001 vs. SHR; ###P < 0.001 vs. WKY).

• 

  Fig. 3. The effect of Sal D on myocardial hypertrophy and apoptosis. (A) WGA was performed to determine cardiomyocytes' cross-sectional area (magnification × 400) (n = 5). (B) The degree of apoptosis in heart tissue was analyzed using TUNEL staining. The nucleus was blue by staining with HOECHST, and positive apoptosis cardiomyocytes were red by staining with TUNEL reagent (magnification × 400) (n = 5). Scale bars 100 μM. (C) Quantification of the fold change of cardiomyocyte size was displayed. (D) Quantification of the percentage of apoptotic cells was exhibited.###, P < 0.001 vs. WKY. ##, P < 0.01 vs. WKY. #, P ​< ​0.05 vs. WKY. ∗∗∗, P ​< ​0.001 vs. SHR. ∗∗, P ​< ​0.01 vs. SHR. ∗, P < 0.05 vs. SHR. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

• 

  Fig. 4. Changes in mitochondrial morphology and mitochondrial function. (A) Ultrastructural analysis of myocardium under 25000 × magnification and the areas of mitochondrion were marked by the red arrow. (B–E) ATP, ADP, AMP concentrations, and energy charge in the myocardium. ###, P < 0.001 vs. WKY. ##, P < 0.01 vs. WKY. #, P ​< ​0.05 vs. WKY. ∗∗∗, P ​< ​0.001 vs. SHR. ∗∗, P ​< ​0.01 vs. SHR. ∗, P < 0.05 vs. SHR. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

• 

  Fig. 5. Functional enrichment analysis of signalling pathways of Sal D treating heart failure. (A) Flow chart of targets searched via TCMSP, BATMAN, and PharmMapper. (B) Significantly enrichment of protein targets in the network diagram.

• 

  Fig. 6. Effect of Sal D on the Ras signalling pathway and PI3K/AKT signalling pathway in the myocardium. Protein expression of (A) p-Ras, Ras, p-Raf, Raf; (B) p-MEK, MEK, p-Erk, Erk; (C) p-PI3K, PI3K, p-Akt, Akt and (D–I) quantization in the myocardium. Results are presented as the mean ​± ​SEM (∗P ​< ​0.05, ∗∗P ​< ​0.01 and ∗∗∗P ​< ​0.001 vs. SHR; ∗P ​< ​0.05, ∗∗P ​< ​0.01 and ∗∗∗P < 0.001 vs. SHR; ###P < 0.001 vs. WKY; ###P < 0.001 vs. WKY).

• 

  Fig. 7. Effect of Sal D on Ras signalling pathway and PI3K/AKT signalling pathway in cardiomyocytes. Protein expression of (A) p-Ras, Ras, p-Raf, Raf; (B) p-MEK, MEK, p-Erk, Erk; (C) p-PI3K, PI3K, p-Akt, Akt and (D–I) their quantization in cardiomyocyte. Results are presented as the mean ​± ​SEM (∗P ​< ​0.05, ∗∗P ​< ​0.01 and ∗∗∗P < 0.001 vs. ANG; #P < 0.05, ##P < 0.01 and###P < 0.001 vs. CON).

• 

  Fig. 8. Effect of Sal D on Ras signalling pathway and PI3K/AKT signalling pathway in cardiomyocytes with Honokiol. Protein expression of (A) p-Ras, Ras, p-Raf, Raf; (B) p-MEK, MEK, p-Erk, Erk; (C) p-PI3K, PI3K, p-Akt, Akt and (D–I) their quantization in the myocardium. Results are presented as the mean ± SEM. (#P < 0.05, ##P < 0.01).