【作者】 刘小美； 方肇勤； 潘志强； 卢文丽； 吴中华； 粱超；

【Author】 LIU Xiao-mei,FANG Zhao-qin,PAN Zhi-qiang,LU Wen-li,WU Zhong-hua,LIANG Chao School of Basic Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China

【机构】 上海中医药大学基础医学院；

【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。重点分析三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Cry1、Per1、Per2和Nrld1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Arntl均明显下调(除邪毒证的Arntl),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Nrld1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。更多还原

【Abstract】 Objective To reveal the expressive characteristic of circadian rhythm pathway in hypothalamic-pituitary-adrenal (HPA) axis of H22 liver cancer mice with different syndromes.Methods Used the quantitative four diagnosis and syndrome differentiation methods to screen and gain normal mice and liver mice with syndrome of obstruction and exuberance of pathogen-toxin(SOEPT) and qi deficiency syndrome(QDS),of which hypothalamus,pituitary,and adrenal gland were get after executed.GeneChip Mouse Exon 1.0 ST Array was used to detect the gene expression profile of every tissue.Then the expressive characteristic of circadian rhythm pathway of these tissues was analyzed.Results After tumorigenesis,the expressive characteristic of circadian rhythm pathway of liver mice with different syndromes was followed,(1) their expression was down-regulated in hypothalamus,especially in it of SOEPT group.And the expression of Cryl,Perl,Per2 and Nr1d1 of QDS group was significantly higher than that of SOPET group.(2) their expression was up- or down-regulated, but had identity between two syndromes.Only Clock gene was down-regulated in SOEPT group among decreased genes and all genes were up-regulated among raised genes except for Perl in QDS group,especially in SOEPT group.And the expression of Perl and Per2 of QDS group was identically lower than that of SOPET group.(3) their expression was up- or down-regulated too,the Clock and Arntl were up-regulated particularly in two syndromes except for Arntl in SOEPT group, especially in QDS group.But the expression of rest genes had bad identity with Clock and Arntl genes of self’s syndrome, and between two syndromes.The expression of Clock and Perl of QDS group was significantly lower than that of SOEPT group,but its Nrldl was higher than that of SOEPT group.Above all,Perl was the common gene which had characteristic of syndrome in HPA axis.Conclusion The circadian rhythm pathway of HPA axis in liver mice was affected in different level. The change of different tissue had relationship with both disease and syndromes,and Perl maybe the key gene which leads the HPA axis develop syndromes difference when tumorigenesis.更多还原