慢性乙型肝炎脾胃湿热证与肝郁脾虚证DNA甲基化的初步研究  

Preliminary Study on DNA Methylation of Chronic Hepatitis B Spleen-Stomach Damp-heat Syndrome and Liver Depression and Spleen Deficiency Syndrome

作　　者：马丽[1,2] 郑秀丽[1] 杨宇[1] 王健[3] MA Li;ZHENG Xiu-li;YANG Yu;WANG Jian(Chengdu University of Traditional Chinese Medicine,Chengdu Sichuan 611137 China;Ningxia Medical University,Yinchuan 750004,China;Pixian Traditional Chinese Medicine Hospital,Chengdu 611730 China)

机构地区：[1]成都中医药大学,四川成都611137 [2]宁夏医科大学,宁夏银川750004 [3]郫县中医院,四川成都611730

出　　处：《时珍国医国药》2021年第7期1671-1675,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81603515)。

摘　　要：目的探讨慢性乙型肝炎脾胃湿热证与肝郁脾虚证的DNA甲基化差异表达的情况。方法招募CHB脾胃湿热证患者7例、肝郁脾虚证患者7例,健康志愿者6例。采集所有受试者清晨空腹无菌环境下静脉血,用ELISA法检测各组间DNA甲基化转移酶1,甲基结合CpG结合蛋白2的表达水平;采用Illumina Bead Array TM Infinium 850K甲基化芯片检测各组间DNA甲基化差异表达的情况。结果两证型组血清DNMT1表达水平与健康对照组相比无显著性差异(P=0.067,P=0.085),肝郁脾虚证组MeCP2表达水平与健康对照组具有显著性差异(P=0.046)。CHB脾胃湿热证特有的高甲基化位点为cg21898358位于基因LILRA3,最显著的低甲基化位点为cg0932788位于基因CHTF18,GO富集结果主要涉及T细胞共刺激、Rho蛋白信号转导、Rho GTPases相关的功能等;CHB肝郁脾虚证组最显著的高甲基化位点为cg09857096位于基因BMPR1B,最显著的低甲基化位点为cg09972436位于基因LCE3C,GO功能富集结果主要涉及树突棘、丝氨酸/苏氨酸激酶、Rac GTPase等。结论 DNA甲基化可能为CHB中医证候的表观遗传学机制之一。Objective To investigate the differential expression of DNA methylation in chronic hepatitis B with spleen-stomach dampness-heat syndrome and liver depression and spleen deficiency syndrome. Methods 7 cases of CHB spleen-stomach dampness-heat syndrome, 7 cases of liver depression and spleen deficiency syndrome were collected, and 6 healthy volunteers were recruited. The venous blood of all subjects was collected in the fasting sterile environment in the morning, and the expression levels of DNA methyltransferase 1, methyl-binding CpG-binding protein 2 were detected by Elisa method;Illumina Bead Array TM Infinium 850 K methylation chip was detected the differential expression of DNA methylation among groups. Results There was no significant difference in the expression level of serum DNMT1 between the two syndrome groups compared with the healthy control group(P=0.067, P=0.085), and the expression level of MeCP2 in the liver depression and spleen deficiency syndrome group was significantly different from the healthy control group(P = 0.046). The hypermethylation site unique to CHB spleen-stomach dampness-heat syndrome is cg21898358 located in gene LILRA3, and the most significant hypomethylation site is cg0932788 located in gene CHTF18. The GO enrichment results mainly involve T cell co-stimulation, Rho protein signal transduction, Rho GTPases Related functions, etc.;The most significant hypermethylation site of CHB liver depression and spleen deficiency syndrome group is cg09857096 located in gene BMPR1 B, and the most significant low methylation site is cg09972436 located in gene LCE3 C. The results of GO function enrichment mainly involve dendritic spines, Serine/threonine kinase, Rac GTPase, etc. Conclusion It is initially confirmed that DNA methylation may be one of the epigenetic mechanisms of CHB syndrome.

关 键 词：慢性乙型肝炎 脾胃湿热证 肝郁脾虚证 DNA甲基化 

分 类 号：R259[医药卫生—中西医结合]