Energy metabolism disorder and myocardial injury in chronic myocardial ischemia with Qi deficiency and blood stasis syndrome based on 2-DE proteomics.

OBJECTIVE: To inquire the characteristic proteins in chronic myocardial ischemia by testing twodimensional electrophoresis (2-DE) map to explore the possible inherent pathological mechanism and the therapeutic intervention of qi deficiency and blood stasis syndrome. METHODS: Ameroid constrictor ring was placed on the first interval of left anterior descending coronary artery to prepare chronic myocardial ischemia model on Chinese miniature swine. Animals were randomly divided into sham group and model group with 10 animals in each group, respectively. The dynamic symptoms observation of the four diagnostic information was collected from 0 to 12 weeks. Echocardiography was employed to evaluate cardiac function and the degree of myocardial ischemia, 2-DE and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) were used to carry out proteomics research on animals. Enzyme-linked immunosorbent assay was applied to identify the relevant differential proteins on chronic myocardial ischemia with qi deficiency and blood stasis syndrome. RESULTS: The preliminary study found that at the 12th week, chronic myocardial ischemia with qi deficiency and blood stasis syndrome model was established stably. Compared with the sham group, there were 8 different proteins down-regulated, 22 proteins up-regulated significantly. After validated by MALDITOF-MS/MS, 11 protein spots were identified. Distinct proteins were mainly associated with energy metabolism and myocardial structural injury, including isocitrate dehydrogenase 3 (NAD+) alpha, NADH dehydrogenase (NAD) Fe-S protein 1, chain A (crystal structure of aldose reductase by binding domain reveals a new Nadph), heat shock protein 27 (HSP27), oxidoreductase (NAD-binding protein), antioxidant protein isoform, cardiac troponin T (cTnT), myosin (myosin light polypeptide), cardiac alpha tropomyosin, apolipoprotein A-I and albumin. CONCLUSION: Down-regulated energy metabolism disorder mediated by NADH respiratory chain and myocardial injury may be the pathogenesis of myocardial ischemia with qi deficiency and blood stasis syndrome. These proteins may be the potential diagnostic marker(s) for qi deficiency and blood stasis syndrome, finally provided new clues for new therapeutic drug target of Chinese medicine.

[Effects of a compound Chinese herbal medicine Yixin Jiedu formula on haemodynamic in rats with heart failure of qi-deficiency and blood stasis syndrome].

OBJECTIVE: To explore the effects of Yixin Jiedu Formula (YXJDF), a compound Chinese herbal medicine, on hemodynamic and B-type natriuretic (BNP) in a rat model of heart failure with qi deficiency and blood stasis syndrome. Moreover, its therapeutic effects in improving the symptoms were also studied. METHODS: The model of heart failure was established by ligation of left coronary artery in rats. Rats were randomly divided into sham-operated group, model group, YXJDF group and positive control (sodium fosinopril tablet) group. On the second day after the operation, rats in different groups received different treatments. Rats in the YXJDF group were treated with YXJDF (9.33g/kg); rats in the positive control group received solution of sodium fosinopril tablet (1.2mg/kg) and rats in the model and the control groups were given an equal volume of saline, respectively. Drugs were delivered by intragastric administration for 28d. Symptoms such as respiratory rate and red-green-blue value of color of the plantar skin were also collected. Then hemodynamic indexes were evaluated and the levels of plasma BNP were examined by enzyme-linked immunosorbent assay (ELISA) in all groups. RESULTS: After 28d, rats in the YXJDF group and the positive control group were more active than rats in the model group. Both YXJDF and sodium fosinopril tablet significantly improved the purple degree of the plantar skin and the respiratory rate. Compared with rats in the sham-operated group, systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) of the model group decreased significantly (P<0.05); rats in the YXJDF group and the positive control group showed significant improvement on SBP, DBP and MAP (P<0.05). In ventricular hemodynamic, left ventricular systolic pressure (LVSP), maximal rate of left ventricular systolic pressure (LV+dP/dt(max)) and maximal rate of left ventricular diastolic blood pressure (LV-dP/dt(max)) of the model group were significantly down-regulated when compared with the sham-operated group (P<0.05); YXJDF and sodium fosinopril tablet increased the LVSP, LV+dP/dt(max) and LV-dP/dt(max) (P<0.05), and thus improved ventricular hemodynamic in rats with heart failure. ELISA results showed that plasma BNP level of the model group was higher than that of the sham-operated group (P<0.05); YXJDF and sodium fosinopril tablet down-regulated the plasma BNP level significantly (P<0.05) compared with the model group. CONCLUSION: YXJDF can strengthen left ventricular contractile force, increase LVSP, LV+dP/dt(max) and LV-dP/dtmax, SBP and SDP, and MAP, and improve hemodynamic indicators in rats with heart failure after myocardial infarction. YXJDF can also relieve the symptoms of qi deficiency and blood stasis syndrome.

[Establishment and evaluation on miniature pig model of ischemic coronary heart disease with qi-deficiency and blood-stasis syndrome].

OBJECTIVE: To study the Chinese medical syndrome characteristics of the miniature pig model of coronary heart disease (CHD) made by chronic myocardial ischemia, and its correlation with heart structure and function. METHODS: Chinese miniature pigs were randomly divided into two groups, 18 in the model group and 8 in the sham-operation group. CHD model was established by placing Ameroid narrow ring in the left anterior descending artery of pigs to induce chronic myocardial ischemia. Characteristics of Chinese medical syndrome in the model were evaluated 8-12 weeks after modeling by observing the general condition, tongue color, activity, hemorrheologic figures, electrocardiogram, coronary angiography, etc., as well as the cardiac structure and function detected by echocardiogram. RESULTS: Compared with the sham-operation group, the modeled pigs showed, at end of the 8th week, pink purple tongue; decreased tail wagging frequency (P < 0.05 or P < 0.01); increased plasma and reduction viscosities (P < 0.05); and raised rigidity index (P < 0.01), with the stenosed diameter of coronary artery reaching 99%. Besides, echocardiogram showed decreased thickness of left ventricular end-systolic stage, end-diastolic stage and inter-ventricular septum end-diastolic stage; lowered ejection fraction and fractional shortening rate (P < 0.05 or P < 0.01), and dilated left ventricular end-diastolic diameter and left ventricular end-systolic diameter. The motion of ventricular wall was decreased orderly in levels from the mitral valve to the papillary muscle to the apex of heart. CONCLUSIONS: CHD pig model made by chronic myocardial ischemia showed a Chinese medical qi-deficiency & blood-stasis syndrome. It indicated that a syndrome transition and conversion progress from blood-stasis to qi-deficiency & blood stasis occurred in the model animals at the 4th to 12th week. The created model provided a reliable basis for further studying rules of syndrome transition and conversion between qi deficiency and blood stasis, as well as on the Chinese medical theory of "blood is mother of qi".

[Analysis of plasma metabonomics of mini-swines with qi deficiency and blood stasis syndrome due to chronic myocardial ischemia].

OBJECTIVE: To study the changes of plasma metabolites of mini-swines with qi deficiency and blood stasis syndrome due to chronic myocardial ischemia and to explore the relationship between plasma metabonomics and qi deficiency and blood stasis syndrome. METHODS: Twenty-two mini-swines were used in this study and were divided into sham-operation group (n=10) and model group (n=12). Ameroid constrictors were placed around the left anterior descending coronary artery of mini-swines in model group to induce chronic myocardial ischemia. Twelve weeks after Ameroid placement, physical signs, coronary angiography and echocardiography were used to evaluate qi deficiency and blood stasis syndrome; gas chromatography-mass spectrometry (GC-MS) and data mining method were used to analyze plasma metabolites of the mini-swines. RESULTS: Twelve weeks after operation, mini-swines in model group were confirmed with qi deficiency and blood stasis syndrome. Principal component analysis (PCA) found that plasma GC-MS spectra of the two groups were significantly different. Compared with sham-operation group, endogenous metabolites in plasma were changed in model group. The levels of lipid metabolites such as glycerol, acetic acid and tricosadiynoic acid, myo-inositol, as well as amino acid metabolite proline were raised, while concentrations of glucose (eg. d-glucose, and a-D-glucoside), amino acid (eg. alanine, phenylalanine, and urea) and lipid metabolites (eg. hexadecanoic acid, propionic acid, butanoic acid, and octadecanoic acid) were declined. CONCLUSION: Significant differences in metabolic spectrum exist in mini-swines with qi deficiency and blood stasis syndrome due to chronic myocardial ischemia and normal mini-swines. Disorders in glucose, amino acid and lipid metabolism result in a deterioration of coronary artery disease; citric acid, glucose, octadecanoic acid, hexadecanoic acid, glycerol and acetic acid are contributory to separation of the healthy from qi deficiency and blood stasis syndrome. These metabolites pattern may be used in clinical diagnosis and treatment of coronary artery disease, and also can provide a new approach to objective research in traditional Chinese medicine syndrome.