Suppression Subtractive Hybridization Screening Of Mouse Kidney Deficiency Syndrome Of Differentially Expressed Genes

Objective: l. To make research foundation for Kidney Deficiency (TCM)related genes and Chinese herbs' targt genes, modem molecular biotechniqueswere used to sift the differentially expressed genes of Kidney Deficiency (TCM)and of those mice treated with Chinese herbs. 2. Theoretic explanations were madefor "Fear impairs Kidney" from different aspects and different levels. Mcthods: l.With the theoretic guide of "Fear impairs Kidney", we combined the methods of"fearng mouse Wth cat "and "suspending mouse over water" to get animal modelof mouse's kidney Deficiency Zheng .At the same time,some mice were treatedwith Chinese herbs, Golden-Box-kidney-Qi pill. 2. The technique of SMARTwas used to get the total full-length cDNAs from the mice's blood cells before andafter they were treated with fear and Chinese herbs. 3. The technique of SSH wasaPPlied in POsitive and negative directions tO acquire the differentially expressedgene profiles under three different states: Kidney Deficiency, Kidney Deficiencybut treated with Chinese herbs, and physiological state. 4. By runningpolyacrylamide gel electroPhoresis, we separaed the cDNA fragments and thenrecovered them. 5. With the works ofamPlification, purification and quantitation.we got the differentially expressed cDNA fragments. 6. In the area of theoreticalresearch, we gave traditional Chinese medicinal explanation to the animal model of"fear impairs Kidney", and then do it from theory of emotional (psychologica1)stress. 7. By using of Chinese Biology-Medicine Disc and Internet, we collectedinformation of "fear" related gene expression. Results: l. Got a animal model ofmouse's Kidney Deficiency. 2. Optimized the protocol of SMART and SSH. 3.Acquired mice's differentially expressed gene profiles under three states and gotsome cDNA fragments of them. Conclusion: 1. The method of "Fear impairsKidney" has practical and theoretical value. 2. Kidney Deficiency Zheng made by"fear' is relatCd with some genes' differentiaI expression, and Chinese herb,Golden-Box-Kidney-Qi pill, can change this result towards which in normalphysiological state. 3. Fear stress is mainly related with the expression of someimmediate early genes and genes of hyPothalamic-pitUitary axis.