垂体特异性转录因子Pou1f1靶基因在不同证候H22荷瘤小鼠的表达特征  

The pou1f1 target genes expression characteristic in pituitary gland of H22 tumor-bearing mice with different TCM patterns

作　　者：潘志强[1] 方肇勤[1] 卢文丽[1] 刘小美[1] 梁超[1] 张园园[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》2017年第2期135-142,共8页Journal of Beijing University of Traditional Chinese Medicine

基　　金：上海市科委国际合作资助项目(No.064307052);上海市进一步加快中医药事业发展三年行动计划资助项目(No.2014-2016年;ZY3-CCCX-3-3010)

摘　　要：目的分析垂体重要功能基因及其垂体特异性转录因子(Pou1f1)靶基因在不同证候荷瘤小鼠的表达差异。方法将H22肝癌腹水细胞接种于190只昆明种小鼠腋下,并设置对照组,采用小鼠标准化四诊信息采集与辨证技术筛选典型的邪毒壅盛证、气虚证、阳气虚证、气阴阳虚证小鼠,并抽提小鼠垂体、下丘脑、肾上腺等组织RNA,采用基因芯片技术检测不同组织基因表达谱,本文重点分析不同证候垂体基因芯片数据。结果与对照组比较,垂体重要功能基因阿片促黑素皮质素原(Pomc)、生长激素(Gh)和催乳素(Prl)在早期的邪毒壅盛证和气虚证中表达下调约10%,而Pomc、Gh、Prl和促甲状腺激素β亚基(Tshb)在中晚期的阳气虚证和气阴阳虚证中表达上调30%以上。Pou1f1在垂体组织中表达最高,呈现组织特异性表达特征;垂体特异性转录因子Pou1f1和Neurod1仅在邪毒壅盛证上调,Tbx19和Egr1在不同证候中均呈现上调特征,Nr4a1、Nr4a2、Pitx1、Gata2、Nr4a3、Foxl2等基因在不同证候中均呈现下调特征。Pou1f1相关靶基因Crem、Tcf4、Rnf41、Lrpprc,Mrps18b、Eif3a、Lars,Arf4、Cep57、Vcpip1,Ptpn6、Myd88、F3,Lrrfip2、Dixdc1与Pou1f1基因表达模式类似,即在邪毒壅盛证上调。结论肿瘤状态下的不同证候小鼠,其垂体重要功能基因表达失常,垂体特异性转录因子Pou1f1及其靶基因在邪毒壅盛证具有类似的表达模式。Objective To study the expression of pituitary key functional genes and their pituitary-specific transcription factor 1 （Poulfl） gene in H22 tumor-bearing mice with different TCM patterns. Methods 190 Kunming mice were injected subcutaneously into the armpit with/422 hepatoma carcinoma cells, and 60 more healthy mice were assigned into the normal group. The mice with pattern of exuberance of patho- genic toxin, asthenia of qi, asthenia of yang and qi, or asthenia of qi, yin and yang were identified by u- sing the method of mice standardization of four diagnosis information arid syndrome differentiation devel- oped by our team. Then RNA was extracted from pituitary gland, hypothalamus and testis and was detec- ted using GeneChip Mouse Exon 1.0 ST Array. And the genetic data of pituitary gland of different patterns were analyzed. Results Compared with normal group, the expression of pituitary key functional genes including Pomc, Gh and Prl were decreased by about 10% in mice with pattern of exuberance of pathogenic toxin or asthenia of qi, which was the pattern of early stage. At the middle or late stage, TCM patters were the pattern of asthenia of yang and qi, or asthenia of qi, yin and yang; Pomc, Gh, Prl and Tshb were upregulated by more than 30%. Poulfl expressed at the highest level in pituitary tissue with the characteristic of tissue specific expression. Poulfl and Neurodl were only upregulated in pattern of exuberance of pathogenic toxin, Tbxl9 and Egrl were upregulated in all patterns, which was just opposite to that of Nr4al, Nr4a2, Pitxl, Gata2, Nr4a3, and Foxl2. Poulfl target genes including Crem, Tcf4, Rnf41, Lrpprc, Mrpsl8b, Eif3a, Lars, Arf4, Cep57, Vcpipl, Ptpn6, Myd88, F3, Lrrfip3 and Dixdcl all expressed in the similar pattern with that of Poulfl, upregulated in pattern of exuberance of pathogenic toxin. Conclusion The abnormal expression of pituitary key functional genes was proved in the tumorbearing mice with different TCM patterns. Pituitary-specific transcription factor 1 with its target

关 键 词：垂体 证候 基因芯片 转录因子 垂体特异性转录因子 阿片促黑素皮质素原:小鼠 

分 类 号：R285.5[医药卫生—中药学]