同病异证H22肝癌小鼠HPA轴昼夜节律通路基因表达特征  

Gene expression characteristics of circadian pathway in hypothalamicpituitary-adrenal axis in mice with H22 liver cancer and different TCM syndromes

作　　者：刘小美[1] 方肇勤[1] 潘志强[1] 卢文丽[1] 粱超[1] 

机构地区：[1]上海中医药大学基础医学院,上海201203

出　　处：《北京中医药大学学报》2013年第8期524-528,共5页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家科技重大专项课题子项目(No.2009ZX09502-018);上海市教委预算内项目(No.2011JW02)

摘　　要：目的揭示同病异证H22肝癌小鼠下丘脑-垂体-肾上腺轴(HPA)昼夜节律通路基因的表达特征。方法采用小鼠计量化四诊和辨证方法筛选获得正常小鼠、早期邪毒壅盛证和气虚证肝癌小鼠,处死取其下丘脑、垂体和肾上腺。以GeneChip Mouse Exon 1.0 ST Array等技术检测各组织的基因表达谱。重点分析3组织各自芯片中昼夜节律通路基因的表达特征。结果肝癌发生后,同病异证肝癌小鼠昼夜节律通路基因在①下丘脑不同程度的表达下降,且以邪毒证为甚。其中气虚证的隐铬1(光解酶样)(Cry1)、阶段基因同源物1(果蝇)(Per1)、阶段基因同源物2(果蝇)(Per2)和核受体亚家族1/组D/成员1(Nr1d1)的表达明显高于邪毒组。②垂体的表达有高有低,但在两证候中表达高低趋势一致,表达下降基因中只有邪毒组的昼夜节律运动输出周期故障(Clock)基因下调明显,而表达升高基因中除气虚证的Per1外,其他均上调明显,且邪毒高于气虚。其中气虚组的Per1和Per2表达明显低于邪毒组。③肾上腺中的表达也有高有低,其中核心基因Clock和芳烃受体核转位因子样(Arntl)均明显下调(除邪毒证的Amtl),且以气虚证为甚。而其他基因的表达与本证候Clock和Arntl基因的表达趋势,以及其在两证候间的表达趋势均显示较差的一致性。其中气虚组的Clock和Per1明显低于邪毒组,而其Nr1d1明显高于邪毒组。以上Per1基因为HPA轴共有的证候特征性基因。结论肝癌小鼠HPA轴的昼夜节律均不同程度受到影响,各组织的变化不仅与疾病本身有关,还具有明显的证候差异,其中Per1可能是HPA轴昼夜节律出现证候差异的关键基因。Objective To reveal the gene expression characteristics of circadian pathway in hypothalamic-pituitary-adrenal （HPA） axis in mice with H22 liver cancer and different TCM syndromes.Methods The mice were screened by using quantitative four examinations and syndrome differentiation,and then divided into normal group,pathogen-toxin exuberance group （pathogen-toxin group） and qi deficiency group.The tissues of hypothalamus,pituitary and adrenal gland were collected from executed mice.The gene expression profiles of these tissues were detected by applying GeneChip Mouse Exon 1.0 ST Array and the gene expression characteristics of circadian pathway were analyzed.Results ① The gene expression of circadian pathway decreased in hypothalamus in varying degrees especially in pathogen-toxin group.The expressions of cryptochrome 1 （photolyase-like） （Cry1）,period homolog 1 （Drosophila） （Per1）,period homolog 2 （Drosophila） （Per2） and nuclear receptor subfamily 1/group D/member 1 （Nr1d1） were significantly higher in qi deficiency group than those in pathogen-toxin group.②The gene expression of circadian pathway was higher or lower in pituitary,but had the same tendency in two syndrome groups.Among the genes with lower expressions,only the gene of circadian locomoter output cycles kaput （Clock） decreased significantly,while among those with higher expressions,except of Per1,others increased significantly especially in pathogen-toxin group.The expressions of Per1 and Per2 were significantly lower in qi deficiency group than those in pathogen-toxin group.③The gene expression of circadian pathway was higher or lower in adrenal gland.The expressions of Clock and aryl hydrocarbon receptor nuclear translocator-like （Arntl） decreased significantly （except of Arntl in pathogen-toxin group） especially in qi deficiency group.The expressions of other genes showed a poor consistency with the expression tendency of Clock and Arnt1 in the same group or between two groups.The expressions of C

关 键 词：同病异证 肝癌 下丘脑 垂体 肾上腺 昼夜节律 小鼠 

分 类 号：R256.4[医药卫生—中医内科学]