基于全基因组甲基化的乙肝后肝硬化典型证候生物学基础观察

Observation of biological foundation on typical syndromes of hepatitis B-caused cirrhosis based on genome-wide DNA methylation profiles

目的:从全基因组DNA甲基化的角度阐释乙肝后肝硬化湿热内蕴证、脾虚湿盛证和肝肾阴虚证证候分类的意义。方法:纳入15例乙肝后肝硬化患者,包括湿热内蕴证、脾虚湿盛证和肝肾阴虚证各5例及6名健康人。采取血液样本,提取DNA做Human Methylation 450K芯片检测及分析。结果:4组之间共有37 957个甲基化位点差异有统计学意义(P<0.05)。与正常组比较,3个证候各有57,68和94个差异甲基化位点,主要集中在基因启动子区。其中,HLA-DRB1和ADAMTS9,NUDT1和YES 1,APOA5和PRKCZ等基因的差异甲基化可能分别与湿热内蕴证、脾虚湿盛证和肝肾阴虚证证候形成有关。结论:DNA甲基化可能为中医证候分类的生物学依据之一。

Objective： To explore the underlying mechanisms of the syndrome classification of internal retention of damp-heat, intense dampness due to spleen deficiency and yin deficiency of liver and kidney of hepatitis B-caused cirrhosis in traditional Chinese medicine based on genome-wide DNA methylation. Methods： 15 cases of hepatitis B-caused cirrhosis patients were divided into 3 groups according to their syndrome difference： syndrome of internal retention of damp-heat（n=5）, syndrome of intense dampness due to spleen deficiency（n=5） and syndrome of yin deficiency of liver and kidney（n=5）, and the control group included 6 healthy cases. Blood samples were taken and DNA was extracted for Human Methylation450 K detection and analysis. Results： 37, 957 loci were statistically differently methylated among 4 groups（P〈0.05）. Compared with the control group, 57, 68 and 94 loci were statistically differently methylated in the syndrome of internal retention of damp-heat, intense dampness due to spleen deficiency and yin deficiency of liver and kidney respectively and most of them located in the promoter region. Among them, differently methylated HLA-DRB1 and ADAMTS9, NUDT1 and YES 1, APOA5 and PRKCZ might associate with the formation of the syndrome of internal retention of damp-heat, intense dampness due to spleen deficiency and yin deficiency of liver and kidney. Conclusion： DNA methylation might be one of the biological basis for syndrome classification.