摘要
目的:探讨脾气虚证大鼠cAMP-PKA信号通路基因表达变化及四君子汤干预作用。方法:24只SPF级雄性SD大鼠,随机分为正常组、脾气虚组、四君子汤组,每组8只,脾气虚证模型采用先饱食1d,再禁食2d,自由饮水,每日游泳至力竭,水温35-37℃,连续15d。造模结束后,通过大鼠的一般状态和生理情况的改变进行模型评价,四君子汤组给予四君子丸混悬液灌胃,正常组和模型组给予等量蒸馏水,连续2周,采用ELISA法检测各组大鼠胃组织cAMP及ATP活性,分别采用实时荧光定量PCR法和Western blot法检测蛋白激酶(PKA)、cAMP反应元件结合蛋白(c REB)、糖原磷酸化酶(GP)、磷酸化酶激酶(PHK)mRNA和蛋白的表达。结果:与正常组大鼠比较,脾气虚证大鼠胃组织cAMP、ATP含量及PKA、GP、CREB、PHK mRNA和蛋白的表达均有所减弱(P<0.01,P<0.05);与脾气虚组大鼠相比,四君子汤组大鼠胃组织cAMP、ATP含量及PKA、GP、CREB、PHK mRNA和蛋白的表达均有所增强(P<0.01,P<0.05)。结论:四君子汤治疗脾气虚证可能与激活胃cAMP-PKA信号转导通路从而影响目的基因的转录有一定的联系。
Objective： To discuss the gene expression in cAMP-PKA pathway of rats with deficiency of spleen qi and intervention effect of Sijunzi Decoction. Methods： Twenty-four SPF male SD rats were randomly divided into three groups including control group, deficiency of spleen qi group and Sijunzi Decoction group with eight rats in each group. The rat models of deficiency of spleen qi were established by using methods of first overfeeding for one day and again fasting for two days, and swimming to exhaustion daily for fifteen days at 35--37℃. The rat models were evaluated by observing the changes in general status and physiological conditions of rats. Rats in control and model group do not accept any treatment, and rats in Sijunzi Decoction group received gavage administration with Sijunzi Pill suspension for twelve weeks. The activity of cAMP and ATP was tasted by using Elisa, and the protein and mRNA expression of cyclic-AMP-dependent protein kinase（PKA）, CREB, glycogen phospholipase（GP） and phosphorylase kinase（PHK） were detected by using Western blot and fluorescent quantitative Realtime PCR methods. Results： The activity of cAMP and ATP, and the protein and mRNA expression of PKA, GP, CREB and PHK in gastric tissue of rats in deficiency of spleen qi group were decreased compared with the control group（P〈0.01, P〈0.05）. Compared with deficiency of spleen qi group, the activity of cAMP and ATP, and the protein and mRNA expression of PKA, GP, CREB and PHK in gastric tissue of rats in Sijunzi Decoction group were increased（P〈0.01, P〈0.05）. Conclusion： The therapeutic effect of Sijunzi Decoction on syndrome of deficiency of spleen qi might be associated with the activation of cAMP-PKA signaling pathway in gastric tissue, and then influenced the gene transcription.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2015年第11期4124-4127,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家重点基础研究发展计划(973计划)项目(No.201133CB531700)~~
