Qishen Yiqi Drop Pill improves cardiac function after myocardial ischemia

Abstract

Myocardial ischemia (MI) is one of the leading causes of death, while Qishen Yiqi Drop Pill (QYDP) is a representative traditional Chinese medicine to treat this disease. Unveiling the pharmacological mechanism of QYDP will provide a great opportunity to promote the development of novel drugs to treat MI. 64 male Sprague-Dawley (SD) rats were divided into four groups: MI model group, sham operation group, QYDP treatment group and Fosinopril treatment group. Echocardiography results showed that QYDP exhibited significantly larger LV end-diastolic dimension (LVEDd) and LV end-systolic dimension (LVEDs), compared with the MI model group, indicating the improved cardiac function by QYDP. (1)H-NMR based metabonomics further identify 9 significantly changed metabolites in the QYDP treatment group, and the QYDP-related proteins based on the protein-metabolite interaction networks and the corresponding pathways were explored, involving the pyruvate metabolism pathway, the retinol metabolism pathway, the tyrosine metabolism pathway and the purine metabolism pathway, suggesting that QYDP was closely associated with blood circulation. ELISA tests were further employed to identify NO synthase (iNOS) and cathepsin K (CTSK) in the networks. For the first time, our work combined experimental and computational methods to study the mechanism of the formula of traditional Chinese medicine.

Figure 1

Representative M-mode echocardiographic view showing measurement of left ventricular internal dimensions for calculation of fractional shortening from the short-axis view of the left ventricle in the myocardial ischemia (MI) model group (A), sham operation group (B), Qishen Yiqi Drop Pill (QYDP) treatment group (C) and Fosinopril treatment group (D), respectively. IVS: Interventricular septum. LVPW: Left ventricular posterior wall. LVIDd: Left ventricular diastolic internal dimension. LVIDs: Left ventricular systolic internal dimension.

Figure 2

600 MHz representative ¹H NMR (δ9.5–5.20, 4.70–0.50) CPMG spectra of serum samples of MI model group (M), sham-operation group (J), QYDP treatment group (Q) and Fosinopril treatment group (F).

Figure 3

(A) PCA score plot of all samples from sham operation group, MI group, Fosinopril treatment group and QYDP treatment group; (B) PCA score plot of MI model group and Sham operation group; (C) PLS-DA score plot of MI model group and Sham operation group; (D) The validation plot derived from MI model group and sham operation group. Solid square for the rats in sham operation group, hollow triangle for the MI group, solid diamond for the Fosinopril treatment group, and cross for the QYDP treatment group.

Figure 4

OPLS-DA score plots (left) and corresponding color-coded correlation coefficient loading plots (right), (A) MI model group vs Sham operation group, (B) MI model group vs QYDP treatment group, (C) MI model group vs Fosinopril treatment group, (D) QYDP treatment group vs Sham operation group, (E) Fosinopril treatment group vs Sham operation group.

Figure 5

600 MHz representative ¹H NMR (δ9.5–5.20, 4.70–0.5) of cardiac muscle samples of MI model group (M), sham-operation group (J), QYDP treatment group (Q) and Fosinopril treatment group (F).

Figure 6. PCA results of cardiac muscle ¹H-NMR spectra.

(A) A score plot of all samples from sham operation group, MI group, Fosinopril treatment group and QYDP treatment group. (B) A loading plot of all samples from the four groups. Solid square for the rats in sham operation group, solid triangle for the MI group, hollow diamond for the Fosinopril treatment group, and cross for the QYDP treatment group.

Figure 7. 9 metabolite biomarkers for QYDP associated with 199 proteins.

Metabolites include Allantoin, Leucine, Alanine, Acetone, Formate, Methionine, Valine, Isoleucine, and Glutamine.

Figure 8. Pyruvate metabolism pathway derived from the identified 20 QYDP-related proteins on the metabolite-protein networks.

The 20 proteins are underlined with green color.