用抑制消减杂交技术研究惊恐致肾虚小鼠差异表达基因  

Application of Suppression Subtractive Hybridization in the Research on Differentially Expressed Genes in the Mice with Kidney Deficiency Induced by Frightening

作　　者：王刚[1] 胡国库[1] 王米渠[2] 张义正[1] 

机构地区：[1]四川大学生命科学学院分子生物学与生物技术四川省重点实验室,四川610064 [2]成都中医药大学中医遗传学研究室,四川610075

出　　处：《北京中医药大学学报》2004年第2期45-48,共4页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目 (No.3 0 17112 6)

摘　　要：目的　用抑制消减杂交技术研究惊恐致肾虚及中药金匮肾气丸治疗后小鼠的差异表达基因。方法　实验分模型组、治疗组和对照组 ,通过“猫吓鼠”法与“水面悬吊应激”法相结合建立小鼠肾虚证动物模型 ,对治疗组小鼠进行中药金匮肾气丸的治疗。应用SMART(SwitchMechanismAt 5endofRNATemplate)技术逆转录并扩增以获取造模及治疗前后小鼠血液的全长总cDNA ;运用正向和反向抑制消减杂交技术获得正反 2个方向、3种状态下 (肾虚、治疗后以及正常生理状态 )的 6组差异表达cDNA片段 ,进而获得了 6个cDNA文库。结果　获得 6个cDNA文库 ,为大规模鉴定、筛选靶基因奠定了基础。结论　惊恐所致肾虚与一些基因的差异表达相关 ,而中药金匮肾气丸能影响这种改变 ,使其差异表达基因谱趋近于正常生理状态。Objective To study the effects of Jinguishenqi Pills (JP) on the differentially expressed genes in the mice with kidney deficiency induced by frightening by the technique of suppression subtractive hybridization (SSH). Methods The experiment included the model group, treating group and control group. The mouse model of kidney deficiency was induced by the methods of using the cat to frighten the mouse and suspending the mouse over the water. The mice in the treating group were treated with JP. SMART technique was used in the reverse transcription and amplification in order to get the total cDNA in full length in the blood cells in the model mice and the model mice prior to and after JP treatment. And then, SSH was applied in both positive and negative directions to gain 6 groups of the differentially expressed cDNA profiles in the model mice, the model mice treated with JP, and in the normal mice. Results 6 Differentially expressed cDNA libraries were acquired in the experiment. Conclusion There is a relationship between the kidney deficiency induced by frightening and the differential expression of some genes, and JP can influence this change in gene expression, inducing the differentially expressed cDNA profiles back to their normal physiological state. The results of the experiment have established a foundation, on which it is able to identify and screen the target genes in a large scale.

关 键 词：恐伤肾 肾虚 SMART技术 抑制消减杂交 基因差异表达 文库 小鼠 

分 类 号：R285.5[医药卫生—中药学]