新疆小白杏杏仁油中生物活性物质免疫调节定量构效关系及作用机制的蛋白质组学研究

Small white apricots oil,recognized as a traditional nutritional supplement and inherited medicament by minority residents,is one kind of unique medical and edible resources in Xinjiang.Currently,some problems,such as the aging of planting structure and the endanger of most wild species,have become the bottleneck restricting the development of small white apricot oil industry.Based on the previous research achievements in the establishment of fingerprints database about biological active substances in small white apricots oil samples,which were collected from major production regions Xinjiang.The immunomodulatory database of different samples has been analyzed and supplied statistically.In this project,the mathematic model,used for the quantitative regression analysis on the relationships of the structure-immunomodulatory activity,will be established by adopting partial least-square regression(PLSR) method,the regression model will be used to quantify the correlations among various active substances(in different samples) and multiple immunology indexes,and then establish the groups of immunocompetent compositions in different samples;Further,the structures of immunocompetent compositions will be identified through isolation and identification joint technologies,including preparative high performance liquid chromatography-mass spectrometry(PHPLC-MS), infrared spectrum(IR) and nuclear magnetic resonance(NMR),and confirm the relationships of the quantitative structure-immunomodulatory activity;Meanwhile, the influence of trial samples on plasma protein compositions of experimental mice will be studied by adopting the technologies of isotope tagging for relative and absolute protein quantitation(iTRAQ)and liquid chromatography-mass spectrometry/ mass spectrometry(LC-MS/MS).In order to mine response protein and explain the molecular immunoregulatory mechanisms of small white apricots oil,the function of differentially expressed proteins and their molecular regulatory networks will also be analyzed.The implementation of this study will provide a good theoretical basis for the optimization of planting resources,the development of wild species and the excavation of specific gene resources.Accordingly,the expected results can accelerate the transformation process of resource advantages into the economic advantages in Xinjiang,this research have profound significance in protecting the characteristics high-quality resources and upgrading the quality of products.

小白杏杏仁油是新疆少数民族长期食用的特色药食两用资源，目前小白杏种植结构老化及野生品种濒危问题已成为制约新疆小白杏杏仁油产业发展的瓶颈。在新疆各产区小白杏杏仁油生物活性物质指纹图谱库构建及其免疫调节数据库补充的研究基础上，本项目采用PLSR方法构建定量构效关系回归分析模型，对不同样本中各类活性物质与多重免疫指标之间的相关性进行量化，建立不同样本功效成分组；采用PHPLC-MS、IR及NMR联合分析技术对功效单体进行结构鉴定，确定功效单体结构与免疫调节之间的定量构效关系；同时采用iTRAQ标记及LC-MS/MS技术研究供试样本对小鼠血浆蛋白组成的影响，通过差异表达蛋白功能及其分子调控网络分析，挖掘与免疫调节相关的应答蛋白，深入解析小白杏杏仁油免疫调节作用分子机制。预期成果将为新疆小白杏种植资源优化、野生抚育及特异基因资源挖掘等方面研究提供坚实的理论依据，对特色资源保护和产品升级具有深远意义。

小白杏杏仁油是新疆少数民族长期食用的特色药食两用资源，目前小白杏种植结构老化及野生品种濒危问题已成为制约新疆小白杏杏仁油产业发展的瓶颈。本项目采用PCA结合PLSR构建定量构效关系回归分析模型，对不同样本中各类活性物质与多重免疫指标之间的相关性进行量化，建立不同样本功效成分组；采用PHPLC-MS、IR及NMR联合分析技术对功效单体进行结构鉴定，确定功效单体结构与免疫调节之间的定量构效关系；采用iTRAQ标记及LC-MS/MS技术研究供试样本对小鼠血浆蛋白组成的影响，通过差异表达蛋白功能及其分子调控网络分析，挖掘与免疫调节相关应答蛋白，解析小白杏杏仁油免疫调节作用分子机制。研究成果如下：.1、针对新疆库车、喀什、阿克苏及和田等主产区21个种植区和市售小白杏杏仁油中生物活性物质进行分析鉴定，对小白杏杏仁油中非甘三酯成分及磷脂类生物活性物质进行补充；.2、通过GC-MS对超临界CO2萃取、机械压榨、超声波辅助提取及索氏提取四种小白杏杏仁油制备样品进行了分析，考察不同方法制备杏仁油样品对氧化应激生物标志物的清除效果。结果表明小白杏杏仁油对4种自由基均存在差异的清除能力，结合PLS1模型明晰了各类物质对样品抗氧化性贡献程度，前期研究成果预测新疆小白杏杏仁油具有调节机体免疫的功能；.3、采用不同剂量的环磷酰胺诱导的方法建立免疫力低下大鼠模型，结合PCA筛选确定9项免疫指标进行后续免疫调节定量购效关系研究；通过免疫力低下试验大鼠模型构建及酶联免疫吸附检测的方法发现，小白杏杏仁油可显著提高免疫力低下大鼠模型的脏器指数，改善相关免疫器官组织病变现象，对大鼠血清及肝脏中IgA、IgM、IgG、IL-12及SOD指标的改善程度达到显著差异水平，对IL-2、TNF-α及GSH-Px及MDA指标的改善程度呈现较显著差异水平；.4、采用iTRAQ标记及LC-MS/MS对小白杏杏仁油干预试验大鼠血清差异表达蛋白进行定量鉴定，并通过表达模式及代谢通路聚类对鉴定获得的差异蛋白进行生化代谢途径和信号转导途径分析。结果表明小白杏杏仁油可能通过调节血清中结合珠蛋白、角蛋白、微管蛋白α-8链、免疫球蛋白重(k)链V-III区、α-1-抗蛋白酶及T-激肽原等蛋白的表达水平，作用于金黄色葡萄球菌感染、吞噬体、致病性大肠杆菌感染、原发性免疫缺陷、补体和凝血级联5条先天性免疫通路而展现其免疫调节作用。