Metabonomic Profiles Delineate the Effect of Traditional Chinese Medicine Sini Decoction on Myocardial Infarction in Rats

Background: In spite of great advances in target-oriented Western medicine for treating myocardial infarction (MI), it is still a leading cause of death in a worldwide epidemic. In contrast to Western medicine, Traditional Chinese medicine (TCM) uses a holistic and synergistic approach to restore the balance of Yin-Yang of body energy so the body’s normal function can be restored. Sini decoction (SND) is a well-known formula of TCM which has been used to treat MI for many years. However, its holistic activity evaluation and mechanistic understanding are still lacking due to its complex components. Methodology/Principal Findings: A urinary metabonomic method based on nuclear magnetic resonance and ultra highperformance liquid chromatography coupled to mass spectrometry was developed to characterize MI-related metabolic profiles and delineate the effect of SND on MI. With Elastic Net for classification and selection of biomarkers, nineteen potential biomarkers in rat urine were screened out, primarily related to myocardial energy metabolism, including the glycolysis, citrate cycle, amino acid metabolism, purine metabolism and pyrimidine metabolism. With the altered metabolism pathways as possible drug targets, we systematically analyze the therapeutic effect of SND, which demonstrated that SND administration could provide satisfactory effect on MI through partially regulating the perturbed myocardial energy metabolism. Conclusions/Significance: Our results showed that metabonomic approach offers a useful tool to identify MI-relate

Bar plots show ¹H NMR relative signal intensities for ten metabolites (A, B) and UHPLC-MS relative signal intensities for thirteen metabolites (C, D) in sham, MI and <i>SND</i>-treated groups.

<p>Data are expressed as mean ± S.D. Significant differences (<i>p</i><0.05) exist between sham and MI for all the metabolites studied.*<i>p</i><0.05 versus MI group.</p

Multivariate statistical analysis of data from the ¹H NMR analysis of urine from sham and MI groups.

<p>(A) PCA scores plot: (▪) sham group, (▴) MI group. (B)The relationship between lambda and deviance using Elastic Net approach (alpha = 0.8).</p

Identification of a selected marker (<i>m/z</i> 180).

<p>(A) Extracted ion chromatogram (EIC) of <i>m/z</i> 180; (B) MS/MS spectrum of the ion; (C) MS/MS spectrum of a commercial standard hippuric acid. The collision energy was 15 V.</p

Typical urinary base peak intensity (BPI) chromatograms of sham rats in (A) ESI⁺ mode and (B) ESI⁻ mode.

<p>Typical urinary base peak intensity (BPI) chromatograms of sham rats in (A) ESI⁺ mode and (B) ESI⁻ mode.</p

Echocardiographic and histological analysis.

<p>(A) M-mode echocardiography in rat models with sham operation, MI and <i>SND</i>-treated groups showing evidence of cardiac failure with chamber dilation in MI rat. <i>SND</i> prevented the development of chamber dilatation in MI rats. (B) Summary data for percentage of ejection fraction (EF) and (C) Summary data for percentage of fractional shortening (FS). Data shown are mean± S.D., n = 7, 6 and 8 for sham, MI, <i>SND</i>-treated groups, respectively. (D) Histologic sections of sham-operated and MI rat hearts, showing gross cardiac dilatation at the 21th day in the MI rats. Treatment of MI rats with <i>SND</i> prevented the development of cardiac remodeling. (E) Myocardial infarct size in sham, MI and <i>SND</i>-treated groups. No infarcted area appeared in sham group. Asterik (*) and pound key (#) represent statistically significant differences compared MI group to sham and <i>SND</i>-treated groups (<i>p</i><0.05), respectively.</p

Representative 600 MHz ¹H NMR spectra of urine from sham (A), MI (B) and <i>SND</i>-treated (C) groups.

<p>Representative 600 MHz ¹H NMR spectra of urine from sham (A), MI (B) and <i>SND</i>-treated (C) groups.</p

Summary of echocardiographic data.

<p>LVEDV, left ventricular end-diastolic volume; LVESV, left ventricular end-systolic volume; LVIDd,left ventricular internal diameter in diastole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; FS, fractional shortening. Data are mean ± S.D.,</p>*<p><i>p</i><0.05 comparing MI and sham animals,</p>#<p><i>p</i><0.05 comparing <i>SND</i>-treatment and MI animals.</p

Identification of significantly differential metabolites in the rat urine by ¹H NMR analysis.

a<p>Letters in parentheses indicate the peak multiplicities: s, singlet; d, doublet; t, triplet; dd, doublet of doublets; q, quarte; ABX, two coupling.</p>b<p>compared to sham group.</p>c<p>compared to MI group. Arrow (↑) indicates relative increase in signal, Arrow (↓) indicates relative decrease in signal, Asterik (*) represents a statistically significant difference(<i>p</i><0.05).</p