[The screening and the functional pathway analysis of differential genes correlated with coronary heart disease of blood stasis syndrome].

OBJECTIVE: To explore the function and target pathway of the correlated differential gene of coronary heart disease (CHD) of blood stasis syndrome (BSS). METHODS: Patients of the genealogical CHD of BSS (group A) and the genealogical CHD of non-BSS (group B), the genealogical non-CHD of BSS (group C), the genealogical healthy subjects (group D), the non-genealogical CHD of BSS (group E), the non-genealogical healthy subjects (group F) were recruited in this study. The differential gene expression spectrums were studied using gene chip technique. The molecular functions of differential genes were analyzed and illustrated by gene ontology (GO) analysis. The differential gene pathways were found out at BioCarta and KEGG. The meaningful target pathways were screened by hypergeometric distribution statistical method. The differential genes were verified using Real-time fluorescent quantitative PCR. RESULTS: (1) By screening the gene chip data (with FC > or =3), we found the expressions of differential genes of CHD of BSS were mainly involved in chemokine, interleukin cytokine, alexin system, matrix metal proteinase system, fibroblastic growth factor, endothelial cell adhesion molecule, and so on. (2) By GO analysis of related differential genes (P < 0.05), we found the molecular functions of differential genes associated with CHD BSS. (3) By BioCarta and KEGG pathway analysis, we found the target pathways of the hereditary correlated differential genes of CHD BSS with significance were mainly involved in inflammation, plaque formation, endothelial injury, and so on. The results of Real-time fluorescent quantitative RT-PCR proved the accuracy of the gene chip. CONCLUSION: The hereditary correlated differential genes of CHD BSS were closely associated with inflammation, plaque formation, and endothelial injury.

[Metabolomics study of the myocardial tissue of rats of cardiac blood stasis syndrome].

OBJECTIVE: To find out the metabolite profile of rats' myocardial tissue of cardiac blood stasis syndrome (CBSS), and to analyze the metabolic pathway of CBSS rats' myocardial tissue by observing the changes of phenotypes intervened by Yangxin Tongmai Recipe (YTR). METHODS: Acute myocardial infarction (AMI) rat model of CBSS was prepared by ligating the left anterior descending coronary artery. Meanwhile, the model was interfered with YTR. The metabolites of rats' myocardial tissue were detected in the model group, the YTR group, the sham-operation group, and the blank control group using GC-MS (8 rats in each group). Changes of metabolite contents were analyzed among different groups using principal component analysis (PCA) and least-square analysis. RESULTS: As for PCA: The results of PCA showed that principal component integral (PCI) of the four groups was mainly distributed in the three regions of oval scatterplot. The factor loading gram showed that contents of glycine, fumaric acid, malic acid, glutamic acid, glucose, phosphoric acid, galactopyranose, lysine were changed in the model group. Analysis of partial least square method: PLS regression model showed that obvious linear correlation existed between the model group and the YTR group, which proved the model was reasonably established. The drug intervention was highly positively correlated with glycine, malic acid, glutamic acid, glucose, highly correlated with urea and butanedioic acid, but negatively correlated with lysine. According to VIP value, each variable was closely correlated with the drug intervention in sequence as malic acid, glutamic acid, glycine, glucose, fumaric acid, urea, galactose, tyrosine, lactic acid, and alanine. Results of variability analysis: Obvious changed variability analysis of metabolite difference showed that 10 metabolites such as glycine, etc. obviously decreased in the model group, showing significant difference when compared with the normal group (P<0.01). Compared with the model group, contents of glycine, fumaric acid, malic acid, glutamic acid, glucose, tyrosine,urea, lactic acid, and alanine, etc. obviously increased after drug intervention (P<0.01). Of them, the increment of malic acid, glumatic acid, tyrosine, and urea was less, showing significant difference when compared with that of the normal group. The mean of lysine was slightly lowered after drug intervention, but with insignificant difference when compared with that of the model group. AMI rats of CBSS was closely correlated with myocardial metabolites such as malic acid, glutamic acid, glycine, glucose, fumaric acid, urea, galactopyranose, lactic acid, alanine, and tyrosine, etc. CONCLUSIONS: The metabolite profile of rats' myocardial tissue showed AMI rat model of CBSS was closely correlated with post-hypoxia glucose metabolism disorder. YTR could effectively intervene this process.

[Detection and analysis on plasma metabolomics in patient with coronary heart disease of Xin-blood stasis syndrome pattern].

OBJECTIVE: To research the plasmic metabolites and metabolic pathway of Xin-blood stasis syndrome (XBSS). METHODS: Plasma metabolic products in patients of coronary heart disease (CHD) with XBSS or non-XBSS and subjects in the control group were identified by gas chromatographic mass spectrometry (GC-MS) type QP2010, the changes of their main elements in different groups were analyzed by principal components analysis (PCA) and partial least squares (PLS) analysis. RESULTS: PCA showed that as compared with that in the control group, in the CHD-XBSS group, contents of lactic acid, beta-hydroxy butanoic acid, urea, oleic acid, octadecanoic acid and arachidonic acid were higher and that of citric acid was lower. PLS analysis showed significant difference between the control group and the other two groups, and the latter two groups tend to be of a same category. The occurrence of XBSS was positively correlated with octadecanoic acid, arachidonic acid, urea, lactic acid and beta-hydroxy, butanoic acid contents, and negatively correlated with oleic acid, L-proline, glycine, and citric acid contents. According to VIP, the degree of correlation between variables with drug interven- tion, from high to low, were ranked as arachidonic acid, octadecanoic acid, lactic acid, urea, beta-hydroxy butanoic acid, linoleic acid, glucose, alanine, oleic acid and proline. Discrepancy analysis on 11 changeful metabolites showed that the contents of arachidonic acid, octadecanoic acid, lactic acid, urea, beta-hydroxy butanoic acid and oleic acid increased in CHD patients, especially in those with XBSS (P < 0.01). In CHD patients, contents of lactic acid, beta-hydroxy butanoic acid, linoleic acid and glucose in patients of XBSS pattern were higher than in non-XBSS pattern (P < 0.01); content of linoleic acid, glucose, alanine and proline decreased in non-XBSS pattern while increased in XBSS pattern. Content of glucose in CHD-XBSS patients was significantly higher than that in the healthy control (P < 0.01). Content of citric acid was lower in CHD patients, and showed significant difference between that in CHD-XBSS patients and healthy control (P < 0.01). CONCLUSIONS: The major plasmic metabolites in CHD-XBSS patients are arachidonic acid, octadecanoic acid, lactic acid, urea, citric acid, beta-hydroxybutyric acid, oleic acid, glucose, and alanine. Analyzed from plasmic metabolite spectrum view, CHD-XBSS is related with lipid metabolism and glyco-metabolism, also with the stress induced by hypoxia and agonia.

[Analysis of the function of vascular endothelial cells in coronary heart disease patients of blood-stasis syndrome].

OBJECTIVE: To explore the function of vascular endothelial cell (VEC) in patients with coronary heart disease (CHD) of Xin-blood-stasis syndrome. METHODS: Some vasoactive substances produced by VEC were detected and analyzed in patients with CHD of or without Xin blood stasis syndrome in group A (n=112) and group B (n=108) respectively, also in patients with non-CHD but of Xin-blood-stasis syndrome in group C (n=110), and healthy persons in group D (n=100), including nitric oxide (NO), endothelin (ET), angiotensin H (Ag II), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule -1 (sVCAM-1). RESULTS: The abnormality degree of ET, Ag II , sICAM-1 and sVCAM-1 in various groups showed such a tendency as group A> group B> group D (P < 0.01 or P < 0.05), while no significant difference in these criteria between group A and group C was shown (P > 0.05). CONCLUSION: The vasoactive substances secreted by VEC are closely related to the formation and progression of CHD, and are likely to be important pathological markers of blood-stasis syndrome in CHD.

[Study on relationship between the polymorphism of angiotensin converting enzyme gene and blood stasis syndrome in patients with coronary heart disease].

OBJECTIVE: To explore the relationship between the insertion/deletion (I/D) polymorphism of angiotensin converting enzyme (ACE), and blood stasis syndrome (BSS) in patients with coronary heart disease (CHD). METHODS: The ACE gene type in 48 patients of CHD of BSS type, 52 CHD patients of non-BSS type and 54 healthy subjects (control) was determined by PCR assay, also levels of endothelin (ET), angiotensin II (Ag II), and nitric oxide (NO) were determined. RESULTS: Occurrence of DD genotype and allele genotype of ACE gene was higher in patients of BSS than that in patients of non-BSS and control (P < 0.01). ET/NO level was higher in patients of BSS than that in control (P < 0.01). ET and Ag II levels in patients of BSS were significantly higher than those in patients of non-BSS (P < 0.05) and control (P < 0.01). Levels of ET/NO and Ag II in subjects with DD genotype in various groups were higher than those in subjects with Ag II or ID genotype, the highest level occurred in patients of BSS with DD genotype, when compared with the other two groups, the difference in Ag II was significant (P < 0.05 and P < 0.01), when compared with control, the difference in ET/NO was significant (P < 0.01). CONCLUSION: DD genotype of ACE gene may be the susceptible gene of CHD in patients of BSS type.