基于方证相关探寻气、血虚证物质基础的蛋白质组学研究  

Proteomic Study on Material Basis of Qi Deficiency Syndrome and Blood Deficiency Syndrome Based on Correlation of Prescription and Syndrome

作　　者：梁华[1] 梁尔新 李奇玮[1] 闫起 王燕[1] 李泽光[2] LIANG Hua;LIANG Erxin;LI Qiwei;YAN Qi;WANG Yan;LI Zeguang(Heilongjiang University of Chinese Medicine, Harbin 150040, China;The First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150040, 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：《中医药学报》2022年第6期16-21,共6页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(81473563)。

摘　　要：目的:以中医方证相关理论为指导,采用iTRAQ蛋白质组学方法观察经典名方补气剂四君子汤、补血剂四物汤对自然衰老模型小鼠肝脏线粒体中蛋白质的影响,挖掘衰老相关的蛋白质标志物,进一步分析四君子汤和四物汤对衰老相关蛋白生物标志物的影响,为方证关系研究提供科学依据。方法:选取40只自然衰老雌性小鼠为研究对象,10只3月龄小鼠为空白组,其他小鼠饲养至16月龄时再将其进行随机分组,分为四君子汤组、四物汤组及模型组,每组10只。四君子汤组、四物汤组灌胃给药剂量为13mL/kg,3月龄空白组与16月龄模型组采用生理盐水灌胃,与给药组等体积比,剂量为13mL/kg。摘取小鼠肝脏组织,并提取线粒体样本,采用同位素标记相对和绝对定量技术,对自然增龄模型小鼠肝细胞线粒体蛋白生物标志物进行研究和分析。结果:空白组与模型组共鉴定出差异蛋白496个;四物汤组有237个标志性蛋白质出现了良性回调;四君子汤组有256个标志性蛋白质出现了良性回调;同时受到四物汤组、四君子汤组影响的蛋白有493个。结论:通过蛋白质组学技术研究发现,补益气血剂对16月龄小鼠肝组织内质网中的蛋白质加工、剪接体及脂肪消化吸收等途径具有调节作用,通过调节这些信号通路的相关蛋白,进而对增龄产生的相关机体功能紊乱起到保护和调节作用。Objective:To observe the changes of protein in liver mitochondria of natural aging model mice intervened by Qi tonifying prescription of Sijunzi Decoction and blood tonifying prescription of Siwu Decoction under the guidance of TCM prescription and syndrome related theory,and to further analyze the effects of Sijunzi Decoction and Siwu Decoction on aging related protein biomarkers,aiming to provide scientific basis for the study on correlation of prescription and syndrome.Methods:40 natural aging female mice were selected as the research objects,10 mice at the age of 3 months were assigned into the blank group,and the other mice were randomly assigned into the Sijunzi Decoction group,the Siwu Decoction group and the model group respectively when they were raised to the age of 16 months,with 10 mice in each group.The dose of the Sijunzi Decoction group and the Siwu Decoction group was 13mL/kg.The 3-month-old blank group and the 16-month-old model group were gavaged with normal saline at the dose of 13 ml/kg.The liver tissue of mice was collected,and the mitochondrial samples were extracted.The relative and absolute quantitative techniques of isotope labeling were used to study and analyze the biomarkers of mitochondrial protein in hepatocytes of naturally aging model mice.Results:496 differential proteins were identified in the blank group and the model group.There were 237 marker proteins with benign callback in the Siwu Decoction group,and there were 256 marker proteins with benign callback in the Sijunzi Decoction group.493 proteins were affected by both Siwu Decoction and Sijunzi Decoction.Conclusion:Through proteomic research,it is found that tonifying Qi and supplementing blood prescription can regulate protein processing and splicing,as well as regulate fat digestion and absorption in the endoplasmic reticulum of liver tissue in 16 month old mice.By regulating the related proteins of these signal pathways,it can protect and regulate the related functional disorders caused by aging.

关 键 词：四君子汤 四物汤 衰老 蛋白质组学 ITRAQ 

分 类 号：R285.5[医药卫生—中药学]