Using suppression subtractive hybridization to research the effects of jinguishenqi pills on the gene expression of the panic-induced kidney deficiency model mice
- VernacularTitle:应用抑制消减杂交研究惊恐致肾虚及金匮肾气丸治疗后小鼠的差异表达基因
- Author:
Gang WANG
;
Guoku HU
;
Yizheng ZHANG
;
Miqu WANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2006;10(39):163-166
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Jinguishenqi pills (JP) can ameliorate the symptoms of kidney deficiency, but the molecular mechanism of the effect is unclear.OBJECTIVE: To observe the differentially expressed genes of panic-induced kidney deficiency model mice treated with JP using suppression subtractive hybridization (SSH).DESIGN: Randomized control animal experiment.SETTING: Institute of Traditional Chinese Medicine Genetics, Chengdu University of Traditional Chinese Medicine; Sichuan Key Laboratory of Molecular Biology and Biotechnology, College of Life Science, Sichuan University.MATERIALS: This experiment was carried out at the Experimental Animal Center of Chengdu University of Traditional Chinese Medicine. Totally 30 BALB/c male adult mice were used in the experiment, and they were randomly divided into model group, JP-treated group and control group,with 10 in each.METHODS: Mice models of kidney deficiency were created in the mice of model group and JP-treated group with the methods of "terrifying mouse with cat" and "suspending mouse over water". Mice of the JP-treated group were intragastrically administrated of JP (produced in Tongrentang Pharmacy Co., Beijing, Z11020147, composed of Guizhi, Fuzi, Shudi, Shanyao,Shanzhuyu, Zexie, Fuling, Danpi, etc) 20-fold as human taken according to the bodyweight once per day. Modeling and administration were conducted in 14 days. Mice of normal control group were fed normally without any extra stimulation. Twenty-four hours after the last administration, 100 μL blood was collected respectively from mice of each group. Switch Mechanism at 5' end of RNA Template (SMART) technique was used in reverse transcription and amplification to obtain the whole length of total cDNA of mice in each group; Forward and reverse SSHs were used to acquire 6 groups of differential expression cDNA segments which were under 3 states(at kidney deficiency, after treatment and at normal physiological status).MAIN OUTCOME MEASURES: Effect of JP on gene expression of kidney deficiency mice RESULTS: ① Differentially expressed cDNA segments were found in each group; The differentially expressed cDNA profile of model mice had restored to that of the normal physiological state by JP treatment. cDNA of the same kind was used as the tester and driver of SSH, and almost all the homologous fragments were subtracted completely, which showed that the subtraction efficiency was very high. ② 6 cDNA libraries were acquired. CONCLUSION: Panic-induced kidney deficiency is related to differential expression of some genes. The gene expression of the kindey deficiency model mice could be affected by JP treatment,making it trend to normal physiological status.
