基于方证相关探寻肾阳虚证物质基础的蛋白质组学研究  

Proteomics on Material Basis of Kidney-Yang Deficiency Syndrome Based on Relevance of Formula and Syndrome

作　　者：梁华[1] 王燕[1] 朱明雪[1] 西旺 李泽光[2] LIANG Hua;WANG Yan;ZHU Mingxue;XI Wang;LI Zeguang(Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China;The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《中华中医药学刊》2020年第12期10-13,I0029,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81473563)。

摘　　要：目的基于中医方证相关理论,采用iTRAQ技术筛选衰老进程中虚证相关差异蛋白变化,以金匮肾气丸干预衰老小鼠,以方测证模式探寻阳虚证的物质基础。方法选用30只自然衰老雄性小鼠为研究对象,3月龄小鼠10只为青年组,剩余小鼠饲养至20月龄后随机分为老龄组和金匮肾气丸组,每组各10只。金匮肾气丸组灌胃给药剂量为10.53 g/kg,其余两组给予等量生理盐水。摘取小鼠肝脏组织,以蛋白滤膜辅助酶解方法进行酶解,之后iTRAQ标记,采用Easy nLC1000和orbitrap Elite液质联用平台对iTRAQ标记样品进行数据采集,经数据库UniProt release 201607筛选,使用Significant B算法利用余补误差函数,以Foldchange>1.2或<0.83,且P<0.05进行差异蛋白的筛选,鉴定出的差异蛋白进行生物信息学分析。结果青年组与老龄组共鉴定出995个差异蛋白;老龄组与金匮肾气丸组共鉴定出674个差异蛋白,发现79个差异蛋白在金匮肾气丸给药后回调。结论金匮肾气丸回调的差异蛋白主要富集在破骨细胞分化、癌症中心碳代谢、氨酰-tRNA生物合成、补体途径和花生四烯酸代谢通路中。Objective Based on the theory of TCM formula and syndrome, iTRAQ technology was used to screen the changes of different proteins related to deficiency syndrome in aging process, Jingui Shenqi pill was used to intervene aging mice, and the material basis of Yang deficiency syndrome was explored by formula and syndrome testing mode. Method Mice liver tissues were extracted and enzymatically hydrolyzed by protein filter membrane-assisted enzymatic hydrolysis method. After that, iTRAQ was labeled. The iTRAQ labeled samples were collected by Easy nLC1000 and Orbitrap Elite liquid chromatography-mass spectrometry platform. The data was screened by UniProt release 201607 database. Significant B algorithm was used to make use of residual error function to Foldchange>1.2 or <0.83 and P<0.05 to carry out differential proteins. The differential proteins were screened and identified for bioinformatics analysis. Results A total of 995 differential proteins were identified in the youth group and the aged group, 674 differential proteins were identified in the aged group and the Jingui Shenqi Pill group, and 79 differential proteins were found to be reversed after Jingui Shenqi Pill was administered. Conclusion The differentially expressed proteins of Jingui Shenqi Pill were mainly concentrated in osteoclast differentiation, central carbon metabolism in cancer, aminoacyl-tRNA biosynthesis, complement and coagulation cascades and arachidonic acid metabolism.

关 键 词：蛋白质组学 金匮肾气丸 阳虚证 方证相关 

分 类 号：R285.5[医药卫生—中药学]