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

Potential Biomarkers in Mouse Myocardium of Doxorubicin-Induced Cardiomyopathy: A Metabonomic Method and Its Application

BACKGROUND: Doxorubicin (DOX) is one of the most potent antitumor agents available; however, its clinical use is limited because of the risk of severe cardiotoxicity. Though numerous studies have ascribed DOX cardiomyopathy to specific cellular pathways, the precise mechanism remains obscure. Sini decoction (SND) is a well-known formula of Traditional Chinese Medicine (TCM) and is considered as efficient agents against DOX-induced cardiomyopathy. However, its action mechanisms are not well known due to its complex components. METHODOLOGY/PRINCIPAL FINDINGS: A tissue-targeted metabonomic method using gas chromatography-mass spectrometry was developed to characterize the metabolic profile of DOX-induced cardiomyopathy in mice. With Elastic Net for classification and selection of biomarkers, twenty-four metabolites corresponding to DOX-induced cardiomyopathy were screened out, primarily involving glycolysis, lipid metabolism, citrate cycle, and some amino acids metabolism. With these altered metabolic pathways as possible drug targets, we systematically analyzed the protective effect of TCM SND, which showed that SND administration could provide satisfactory effect on DOX-induced cardiomyopathy through partially regulating the perturbed metabolic pathways. CONCLUSIONS/SIGNIFICANCE: The results of the present study not only gave rise to a systematic view of the development of DOX-induced cardiomyopathy but also provided the theoretical basis to prevent or modify expected damage

Molecular Modeling Study of Chiral Separation and Recognition Mechanism of β-Adrenergic Antagonists by Capillary Electrophoresis

Chiral separations of five β-adrenergic antagonists (propranolol, esmolol, atenolol, metoprolol, and bisoprolol) were studied by capillary electrophoresis using six cyclodextrins (CDs) as the chiral selectors. Carboxymethylated-β-cyclodextrin (CM-β-CD) exhibited a higher enantioselectivity power compared to the other tested CDs. The influences of the concentration of CM-β-CD, buffer pH, buffer concentration, temperature, and applied voltage were investigated. The good chiral separation of five β-adrenergic antagonists was achieved using 50 mM Tris buffer at pH 4.0 containing 8 mM CM-β-CD with an applied voltage of 24 kV at 20 °C. In order to understand possible chiral recognition mechanisms of these racemates with CM-β-CD, host-guest binding procedures of CM-β-CD and these racemates were studied using the molecular docking software Autodock. The binding free energy was calculated using the Autodock semi-empirical binding free energy function. The results showed that the phenyl or naphthyl ring inserted in the hydrophobic cavity of CM-β-CD and the side chain was found to point out of the cyclodextrin rim. Hydrogen bonding between CM-β-CD and these racemates played an important role in the process of enantionseparation and a model of the hydrogen bonding interaction positions was constructed. The difference in hydrogen bonding formed with the –OH next to the chiral center of the analytes may help to increase chiral discrimination and gave rise to a bigger separation factor. In addition, the longer side chain in the hydrophobic phenyl ring of the enantiomer was not beneficial for enantioseparation and the chiral selectivity factor was found to correspond to the difference in binding free energy

Metabolic Profiling Provides a System Understanding of Hypothyroidism in Rats and Its Application

<div><h3>Background</h3><p>Hypothyroidism is a chronic condition of endocrine disorder and its precise molecular mechanism remains obscure. In spite of certain efficacy of thyroid hormone replacement therapy in treating hypothyroidism, it often results in other side effects because of its over-replacement, so it is still urgent to discover new modes of treatment for hypothyroidism. <em>Sini</em> decoction (<em>SND</em>) is a well-known formula of Traditional Chinese Medicine (TCM) and is considered as efficient agents against hypothyroidism. However, its holistic effect assessment and mechanistic understanding are still lacking due to its complex components.</p> <h3>Methodology/Principal Findings</h3><p>A urinary metabonomic method based on ultra performance liquid chromatography coupled to mass spectrometry was employed to explore global metabolic characters of hypothyroidism. Three typical hypothyroidism models (methimazole-, propylthiouracil- and thyroidectomy-induced hypothyroidism) were applied to elucidate the molecular mechanism of hypothyroidism. 17, 21, 19 potential biomarkers were identified with these three hypothyroidism models respectively, primarily involved in energy metabolism, amino acid metabolism, sphingolipid metabolism and purine metabolism. In order to avert the interference of drug interaction between the antithyroid drugs and <em>SND</em>, the thyroidectomy-induced hypothyroidism model was further used to systematically assess the therapeutic efficacy of <em>SND</em> on hypothyroidism. A time-dependent recovery tendency was observed in <em>SND</em>-treated group from the beginning of model to the end of treatment, suggesting that <em>SND</em> exerted a recovery effect on hypothyroidism in a time-dependent manner through partially regulating the perturbed metabolic pathways.</p> <h3>Conclusions/Significance</h3><p>Our results showed that the metabonomic approach is instrumental to understand the pathophysiology of hypothyroidism and offers a valuable tool for systematically studying the therapeutic effects of <em>SND</em> on hypothyroidism.</p> </div

Sample determination of total triiodothyronine (T₃) and total thyroxine (T₄) in rat serum for the surgery-induced hypothyroid groups (mean±S.D.)^a (n = 8).

a<p>The unit of T₃ and T₄ is ng/mL.</p>**<p><i>p</i><0.01 (compared to Sham group),</p>***<p><i>p</i><0.001(compared to Sham group).</p>#<p><i>p</i><0.05 (compared to Hypo group),</p>##<p><i>p</i><0.01 (compared to Hypo group),</p>###<p><i>p</i><0.001 (compared to Hypo group).</p

Typical urine total ion current (TIC) chromatograms in positive ionization mode (A), negative ionization mode (B).

<p>Typical urine total ion current (TIC) chromatograms in positive ionization mode (A), negative ionization mode (B).</p

The Body Weight (BW) profiles during the model and treatment periods.

<p>(box represents Sham group, triangle represents Sham + <i>SND</i> group, dot represents Hypo group, diamond represents Hypo + <i>SND</i> group, inverted triangle represents Hypo + T₄ group).</p

Comparison of different ions in urine of Sham group, Sham + <i>SND</i> group, Hypo group, Hypo + <i>SND</i> group and Hypo + T₄ group after 4-week treatment.

<p>* <i>p</i>-Value<0.05 compared to Sham group, ** <i>p</i>-Value< 0.01 compared to Sham group, *** <i>p</i>-Value<0.001 compared to Sham group; ^#<i>p</i>-Value<0.05 compared to Hypo group, ^##<i>p</i>-Value<0.01 compared to Hypo group, ^###<i>p</i>-Value<0.001 compared to Hypo group.</p

PLS-DA scores plot (top panel), loading plot (middle panel) and S-plot (bottom panel) of the UHPLC/TOF-MS spectral from control group (box) and hypothyroid group (dot).

<p>(<b>A</b>: control group <i>vs</i> MMI group; <b>B</b>: control group <i>vs</i> PTU group; <b>C</b>: control group <i>vs</i> thyroidectomy-induced hypothyroid group).</p

The integrative plot of the metabolites and the relevant pathways changing for thyroidectomy-induced hypothyroidism in circulation system.

<p>Metabolites with red dashed area present significant increase in Hypo group compared to Sham group. Metabolites with blue dashed area present significant decrease in Hypo group compared to Sham group.</p