摘要
目的:通过检测和分析慢性乙型病毒性肝炎(以下简称乙肝)肝肾阴虚证、湿热蕴结证患者的差异基因表达谱,探讨乙肝中医证候分型与基因表达之间的关联。方法:从上海龙华医院肝科选择乙肝患者中诊断为肝肾阴虚证和湿热蕴结证者各3例,并选择3例健康者作为对照。采集乙肝患者和健康者的外周血样本,提取白细胞中的总RNA,运用基因芯片技术检测外周血基因表达情况,并对其表达谱进行比较。另外选择肝肾阴虚证和湿热蕴结证患者各10例和健康者10例,使用实时定量逆转录酶-聚合酶链式反应法验证部分基因的表达情况。结果:在乙肝肝肾阴虚证和湿热蕴结证患者之间、乙肝患者与健康者之间,基因表达谱都存在明显的差异表达。两种证型患者之间共有403个差异表达的基因,其中,239条基因表达有显著性差异(|差异倍数|≥2,P<0.05),基因表达上调的有142条,下调的有97条。肝肾阴虚证单独调变的基因主要与过氧化物酶的活性和干细胞的维持等相关,湿热蕴结证单独调变的基因主要与细胞因子、免疫应答和炎症反应等相关。实时定量逆转录酶-聚合酶链式反应验证ATP结合盒亚家族C成员3、过氧化物酶增殖体激活受体α等基因的表达与基因芯片检测结果趋势一致。结论:乙肝肝肾阴虚证和湿热蕴结证患者白细胞中基因表达存在差异,提示乙肝中医临床辨证分型具有分子生物学基础,与基因差异表达密切相关。
OBJECTIVE：To investigate the differential gene expression profile in two typical traditional Chinese medicine （TCM）syndromes of patients with chronic hepatitis B（CHB）,and to find the relationship between TCM syndromes and gene expressions. METHODS：Patients with CHB were collected from Department of Liver Diseases of Longhua Hospital in Shanghai and diagnosed as dual deficiency of liver and kidney yin syndrome（n=3）or accumulation of dampness heat syndrome（n=3）.Three healthy volunteers were used as a control.Blood samples were collected before treatment.After total RNA was isolated from leukocytes,the gene expression profiles were detected by microarray.For confirming the gene expressions,a further 10 patients with dual deficiency of liver and kidney yin syndrome or accumulation of dampness heat syndrome and 10 healthy volunteers were enrolled.Their peripheral blood was used to test the gene expression by real-time reverse transcription- polymerase chain reaction（RT-PCR）which was differently expressed by microarray. RESULTS：Microarray analysis identified that there were significant differences in gene expression between patients with dual deficiency of liver and kidney yin syndrome and patients with accumulation of dampness heat syndrome,as well as between CHB patients and healthy volunteers.Patients with dual deficiency of liver and kidney yin syndrome and accumulation of dampness heat syndrome were fully distinguished by clustering of 403 differentially expressed genes（P0.05）.A total of 239 genes were significantly differentially expressed（absolute value of foldchange≥2,P0.05）,among which 142 were up-regulated and 97 were down-regulated.Genes specially regulated by the dual deficiency of liver and kidney yin syndrome were mainly related to peroxide activity and stem cell maintenance;genes specially regulated by the accumulation of dampness heat syndrome were mainly related to cytokines,immune and inflammatory responses.ATP- binding cassette,sub-family C（CFTR/MRP）,member 3,p
出处
《中西医结合学报》
CAS
2012年第7期751-756,共6页
Journal of Chinese Integrative Medicine
基金
上海市教育委员会重点学科建设项目资助(No.J50301)
上海市教育委员会E-研究院中医内科建设计划资助项目(No.E03008)
关键词
肝炎
乙型
慢性
证候
肝肾阴虚
湿热
基因
基因表达谱
非治疗性人类实验
hepatitis B
chronic
syndromes
liver-kidney yin deficiency
dampness-heat
gene
gene expression profiling
nontherapeutic human experimentation
