基于DNA甲基化的慢性乙型病毒性肝炎脾胃湿热证与肝郁脾虚证的KEGG通路富集分析

KEGG pathway enrichment analysis of spleen-stomach dampness heat syndrome and liver depression and spleen deficiency syndrome in chronic hepatitis B based on DNA methylation

目的:观察慢性乙型病毒性肝炎(CHB)脾胃湿热证与肝郁脾虚证DNA甲基化差异位点基因在KEGG通路的富集情况。方法:纳入CHB脾胃湿热证7例,CHB肝郁脾虚证7例,健康对照者6例。采集受试者空腹静脉血5mL,及时分离血清提取DNA,采用Illumina BeadArray TM Infinium 850K甲基化芯片检测,筛选各组间对应KEGG数据库中的差异甲基化基因数及KEGG通路的富集情况。结果:与健康对照者比较,CHB组、CHB脾胃湿热证组、CHB肝郁脾虚证组及CHB两证候组间均有差异甲基化基因及KEGG通路的富集。CHB脾胃湿热证组主要涉及泛素介导的蛋白水解作用、肾脏上皮肾细胞癌等通路;CHB肝郁脾虚证组主要涉及类固醇生物合成、嘌呤代谢等通路。结论:CHB不同中医证候的差异甲基化基因涉及多条KEGG通路,且具有一定的趋向性。

Objective: To observe the accumulation of DNA methylation genes in KEGG pathway in spleen-stomach dampness heat syndrome(SSDHS) and liver depression and spleen deficiency syndrome(LDSDS) of chronic hepatitis B(CHB). Methods: Seven cases SSDHS of CHB, 7 cases LDSDS of CHB and 6 healthy controls were included. Collect 5 mL of fasting venous blood from the subject, separate the serum in time to extract DNA, and use the Illumina BeadArray TM Infinium 850 K methylation chip to detect the number of differentially methylated genes in the KEGG database and the enrichment of the KEGG pathway between each group. Results: Compared with the healthy control group, CHB group, CHB SSDHS group, CHB LDSDS group and CHB two syndrome groups all had differential methylation genes and KEGG pathway enrichment. The SSDHS of CHB mainly involved ubiquitin-mediated proteolysis, renal epithelial renal cell carcinoma and other pathways. The LDSDS of CHB mainly involved steroid biosynthesis, purine metabolism and other pathways. Conclusion: The methylated genes of different TCM syndromes in CHB involve multiple KEGG pathways and have certain tendency.