Differential Expression Profile Analysis Of Coronary Heart Disease And Angina Pectoris With Blood Stasis Syndrome And Methylation Of IL6 Gene

Background:The Traditional Chinese Medicine（TCM） pattern Blood Stasis Syndrome（BBS） of Coronary Heart Disease（CHD） is associated with the pathogenesis of microcirculation disturbance, hypercoagulable state, blood platelet activation and adhesion and gathering, thrombogenesis, immune dysfunction. Meanwhile, the specific mRNA, miRNA and lncRNA of BBS has also been discovered for years. However, the mechanism of BBS has not been illustrated clearly and the unique biomarker of BBS for quantitative diagnosis and molecular therapeutic target have not been discovered and applied for clinic.Objective:Analyze the network and key regulator from gene expression profile of BBS based on bioinformatics. Explore the potential influence of DNA methylation on gene expression profile of BBS on account of epigenetics.Method:We searched the lectures concerned with BBS and results about gene expression profile on the databases of PubMed, CNKI, VIP, WanFang and SinoMed. We used the DAVID platform to annotate the genes and functional enrichment analysis and pathway analysis. GeneCards platform was used to find the distribution of targeted gene in subcellular structure and STRING network of expressed proteins. Blood samples were obtained from 6 patients with BBS and 6 patients with non BBS. Genomic DNA was extracted from the blood samples and underwent bisulfate treatment using the EZ DNA methylation kit （Zymo Research, Irvine, CA, USA） according to the manufacturer’s instruction. The annotation of the targeted DNA was searched on UCSC and the design of primers was accomplished by Epidesigner. After PCR, SAP treatment and transcription by T7 DNA polymerase, the Sequenom MassARRAY platform（Sequenom, SanDiego, CA, USA） was used to quantitatively analyze the methylation status of the targeted region.Results:The Go analysis was enriched on the pathway of inflammatory response. The BIOCARTA analysis was enriched on the pathway of Cells and Molecules involved in local acute inflammatory response, involving the gene of L6、 ITGB2、 C3. BBS samples was exhibited higher than that in non BBS group at a total of 7 CpG sites from Region IL6₁（from-1118bp to-826bp）, whereas no significant difference in methylation level between the two groups was observed in Region IL6₂（-1471bp to-1184bp）.Conclusion:The BBS of CHD is closely related to the inflammatory response, In the sequence of 2000bp before the transcriptional start site of the gene IL6, BBS samples was exhibited higher than that in non BBS group at a total of 7 CpG sites from Region IL6₁（from-1118bp to-826bp）, whereas no significant difference in methylation level between the two groups was observed in Region IL6₂（-1471bp to-1184bp）. This research pointed out the regulation of BBS by DNA methylation to a certain degree.