慢性乙型肝炎和肝硬化“异病同证”的全基因组甲基化分析

Genome-Wide Methylation Analysis in the Same Syndrome Between the Chronic Hepatitis B and Hepatitis B-Related Cirrhosis

目的:本文从全基因组DNA甲基化的角度探讨慢性乙型肝炎(慢乙肝)和慢乙肝后肝硬化(肝硬化)湿热内蕴证、肝郁脾虚证和肝肾阴虚证异病同证的生物学基础。方法:采取慢乙肝及肝硬化湿热内蕴证、肝郁脾虚证和肝肾阴虚证患者和健康志愿者外周血样本,提取DNA进行Human Methylation450K芯片检测及分析。结果:慢乙肝和肝硬化异病同证湿热内蕴证特异性差异甲基化位点有9个,覆盖9个基因;肝郁脾虚证有30个,覆盖20个基因;肝肾阴虚证有22个,覆盖14个基因。其中,与正常组相比,湿热内蕴证中KCTD2和NAV1,肝郁脾虚证中LGR6和SH2D4B及肝肾阴虚证中CYP2E1、PCSK6、DEXI、HIST1H3B和SULT1C2的差异位点甲基化程度变化较大(|Delta Beta|>0.15)。结论:慢乙肝与肝硬化基因甲基化可能与其湿热内蕴证、肝郁脾虚证和肝肾阴虚证的形成有关。

This study aimed to explore the biological basis of the same syndrome of Traditional Chinese Medicine（TCM） in different diseases, involving dampness and heat excess（DH） syndrome, liver stagnation and spleen deficiency（LSSD） syndrome and liver- and kidney-yin deficiency（LKYD） syndrome in chronic hepatitis B（CHB） and hepatitis B-related cirrhosis（LC）. The CHB and LC patients with DHS, LSSDS and LKYDS（10 cases apiece） and healthy volunteers（6 cases） were enrolled. Blood samples were prepared and DNAs were extracted for Human Methylaiton450 K detection and analyses. As a result, 9 loci（covering 9 genes） were specifically and differently methylated in DH syndrome in both the two diseases, while 30 loci（covering 20 genes） in LSSD syndrome and 22 loci（covering 14 genes） in LKYD syndrome. Among the genes, KCTD2 and NAV1 in DH syndrome, LGR6 and SH2D4 B in LSSD syndrome, and CYP2E1, PCSK6, DEXI, HIST1H3 B and SULT1C2 in LKYD syndrome were significantly differently methylated compared with the healthy group（｜Delta Beta｜〉 0.15）. In conclusion, methylation level alternation may involve in the formation of DH, LSSD and LKYD syndromes in CHB and LC.