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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 differentially expressed genes in mice after frightened phlegm-induced kidney deficiency and Chinese medicine Jinkuishenqiwan by suppression subtractive hybridization. Methods The experiment was divided into model group, treatment group and control group. Animal models of Kidney Deficiency Syndrome were established by combining “Cat Scare” method with “Water Suspension Stress” method. The mice in treatment group were treated with Jing Kui Shen Qi Wan treatment. Reverse transcription and amplification were performed using SMART (Switch Mechanism of 5 end of RNATemplate) technology to obtain the full-length cDNA of the blood of the model mice and the mice before and after treatment. The forward and reverse inhibition subtractive hybridization techniques were used to obtain positive and negative directions. Under the three conditions Kidney deficiency, after treatment and normal physiological state) of 6 groups of differentially expressed cDNA fragments, and then obtained 6 cDNA libraries. Results Six cDNA libraries were obtained, which laid the foundation for large-scale identification and screening of target genes. Conclusions Kidney deficiency caused by panic is related to the differential expression of some genes, whereas Jinkuishenqiwan can affect this change and make its differentially expressed gene spectrum approach the normal physiological state.