Differential gene expression profiles in coronary heart disease patients of blood stasis syndrome in traditional Chinese medicine and clinical role of target gene.

OBJECTIVE

To investigate the differential gene expression profiles in coronary heart disease (CHD) patients of blood-stasis syndrome (BSS) by oligonucleotide microarray technique, and the clinical significance of target gene.

METHODS

Subjects were assigned to CHD patients with BSS (n=8), CHD patients without BSS (n=8), and BSS patients without CHD (n=8) based on coronary angiography and the diagnostic criteria of BSS. The sex- and age-matched healthy volunteers (n=8) were enrolled as the control group. Venous blood samples were collected for RNA extraction; Test-3 chip was employed to examine the quality of samples. Then, the samples were hybridized with Affymetrix U133 Plus 2.0 array to compare the gene expression profiles among the four groups. Gene-array scanner and gene chip operating software were applied to screen out hybridization signals and analyze gene expression, respectively. Based on the comparison of the samples of the four groups, the differential genes related with CHD and BSS were analyzed with Gene Ontology (GO) and pathway, and target genes selected were confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR). Thirty CHD patients with BSS were selected according to the former criteria and 40 healthy as controls. The serum concentration of interleukin-8 (IL-8) was determined by double-antibody sandwich avidin-biotin peroxidase complex enzyme-linked (ABC-ELISA).

RESULTS

A total of 107 differential genes were found being associated with CHD, including 48 up-regulated genes and 59 down-regulated genes. Among these 107 differential genes, 14 genes (13.1%) were found related to inflammatory reaction and immune response through GO analysis. In the pathway analysis, 4 of 15 conspicuous pathways were referred to the inflammation and immune response. Among 48 differential genes related to BSS, 26 genes were up-regulated, and 22 were down-regulated. Five of the 48 genes (10.4%) and 5 of 10 significant pathways were involved in inflammation and immunity. The results of real-time RT-PCR proved the accuracy of the gene chip. The patients have markedly higher level of serum IL-8 compared to the controls (P<0.05).

CONCLUSION

The correlation of inflammatory- and immune-related genes with CHD patients of BSS was revealed at the level of nucleic acid, and the target gene IL-8 may play a role in the pathobiology of CHD with BSS.