肝郁脾虚证模型大鼠肝脏的蛋白组学研究  

Proteomics analysis on liver proteins in rats with syndrome of liver qi stagnation and spleen deficiency

作　　者：李聪[1] 谢鸣[1] 赵荣华[1,2] 孙伟[3] 郭正光 

机构地区：[1]北京中医药大学,北京100029 [2]中国中医科学院中药研究所,北京100700 [3]中国医学科学院协和基础研究所分子生物学实验中心,北京100730

出　　处：《中华中医药杂志》2016年第10期4211-4215,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81173193)~~

摘　　要：目的:运用蛋白组学技术探查中医肝郁脾虚证模型大鼠肝脏的分子生物学内涵。方法:大鼠随机分为正常对照组和肝郁脾虚证模型组,每组10只。模型组采用慢性束缚+饮食失节+过度疲劳的方法制模,正常组不予任何处理,连续28d。实验第29天,低温条件下剖取肝脏同一部位的组织,生理盐水冲洗后,进行蛋白质提取、酶切后i TRAQ标记、高p H值反相液相色谱分离、液-质联用分析,获得质谱检测数据;经反相数据库评估、蛋白质鉴定及定量后,确定模型组与正常对照组的差异蛋白谱;运用IPA软件分析获得模型组差异蛋白所涉及的功能、主要信号传导通路及关联蛋白网络。结果:肝郁脾虚证模型大鼠肝脏差异蛋白共有287个,其中上调64个,下调223个;明显变化的信号传导通路有11条,重要的蛋白相互作用通路2个。结论:肝郁脾虚证模型大鼠肝脏存在多个蛋白的异常表达,其功能涉及清除异物毒物、胆固醇及多种激素代谢、尼古丁降解、免疫调节等信号通路,提示中医肝郁脾虚证生物学内涵可能涉及肝脏上述分子通路的调控异常。Objective： To explore the molecular biology connotation of liver in rats with stagnation of liver qi and spleen deficiency syndrome（GYPX） by proteomics analysis. Methods： Rats were randomly divided into the control group and GYPX model group, 10 rats of each group. The GYPX model was induced by chronic restraint stress, improper diet, and excess fatigue for 28 days. After modeling, the liver of rats were collected; then undergone the protein digestion, i TRAQ labeling, Offline reverse-phase liquid chromatography（RPLC） separation, Online LC/MS/MS analysis, to obtain the MS detection data; after the evaluation of the reverse phase database, the identification and quantification of proteins, the differential protein profiles between the model group and control group were determined; then, the function of differential proteins of the model group, main signaling pathways and associated protein network that differentially expressed proteins were analyzed with IPA software. Results： There were 287 differentially expressed proteins in the GYPX group, of which 64 proteins were up-regulated and 223 were downregulated. The significant change signaling pathways were 11 and the main protein interaction pathways were 2 in the GYPX group. Conclusion： There are multiple proteins differentially expressed in the liver of GYPX model rats, the restraining functions include signaling pathways of foreign matter removal, cholesterol and a variety of hormone metabolism, nicotine degradation and immune regulating. It prompts that the connotation of stagnation of liver qi and spleen deficiency syndrome may involve the above molecular pathways of the liver.

关 键 词：肝郁脾虚证 大鼠模型 肝脏 蛋白组学 

分 类 号：R-332[医药卫生] R228