应用液质联用技术研究脾虚大鼠血清代谢物谱群特征  

Study on Serum Metabolite Profiling in Pi-deficiency Rats Based on LC-MS Technique

作　　者：贾连群[1] 甄毕贤[1] 徐荧[1] 杨关林[1] 

机构地区：[1]辽宁中医药大学中医脏象理论及应用教育部重点实验室,沈阳110847

出　　处：《中国中西医结合杂志》2016年第3期359-365,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家重点基础研究发展计划("973"计划)资助项目(No.2013CB531704);国家自然科学基金青年基金项目(No.81202834)

摘　　要：目的应用液质联用技术（LC-MS）研究脾气虚、脾阳虚大鼠血清代谢物谱群变化特征,并从小分子代谢物水平探讨脾虚证候本质。方法 21只SPF级雄性SD大鼠,随机分为正常组、脾气虚组、脾阳虚组,每组7只。脾气虚证模型造模方法采用先饱食1天,再禁食2天,自由饮水,每日游泳至力竭,水温35~37℃,连续15天。脾阳虚证造模方法采用脾气虚证加灌服20%番泻叶水浸剂（2 m L/100 g）,早晚各1次,连续1周。在造模结束后,根据大鼠的一般情况、体重变化及肛温的改变进行模型评价,运用LC-MS技术对大鼠进行血清代谢组学检测,应用正交偏最小二乘判别分析研究组间代谢物谱图差异,并通过变量重要性投影（OPLS-DA）选取血清中与证型相关的生物标志物。结果与造模前比较,脾气虚组和脾阳虚组大鼠的体重均有显著降低,两组体重差值与正常组比较,差异有统计学意义（P〈0.01）,脾阳虚组较脾气虚组体重值降低更明显（P〈0.05）;与正常组比较,脾气虚组和脾阳虚组大鼠的肛温下降,脾阳虚组较脾气虚组降低更多（P〈0.05,P〈0.01）。与正常组比较,脾气虚组与脾阳虚组PC（19∶0）/PE（22∶0）、PC（17∶0）/PE（20∶0）、癸酸、油酸、硬脂酸、琥珀酸、延胡索酸、苹果酸、葡萄糖含量升高,花生四烯酸、亚麻酸、十二碳烯酸、雄酮、4-庚酮、DHAP（6∶0）、尿苷等含量降低。与脾气虚组比较,脾阳虚组PC（22∶1）、PC（22∶6）、PE（18∶0）/PC（15∶0）、视黄醇及脱氧胞苷相对含量显著升高,PC（18∶1）、PC（19∶3）、PC（20∶3）、PC（17∶0）/PE（20∶0）、PC（19∶1）/PE（22∶1）、PC（19∶0）/PE（22∶0）、PC（17∶1）/PE（20∶1）、PC（16∶1）/PE（19∶1）、胆酸、马尿酸、糠酸、十一烷二甲酸、棕榈油酸、羟基硬脂酸、二十碳三烯酸、苯丙氨酸、酪氨酸、谷氨酸、丝氨酸、氨基甲酰基天冬氨酸、Objective To explore metabolite profiling changes in serum of rats with pi-qi deficiency syndrome（ PQDS） and pi-yang deficiency syndrome（ PYDS） based on liquid chromatograph-mass spectrometer（ LC-MS） technique,and to explore the essence of Pi-deficiency syndrome（ PDS） from small molecule metabolite level. Methods Totally 21 male SD rats of SPF grade were randomly divided into three groups,the normal control group,the PQDS group,and the PYDS group,7in each group. Rats in the PQDS group overate for 1 day and fasted for 2 days. They drank freely and then swam to be exhausted in water at 35 ℃- 37 ℃ for 15 successive days. The PYDS model was established by the same method for PQDS rats plus drenching 20% Folium sennae water extract（ 2 m L /100 g）,once in the morning and once in the evening for one successive week. After modeling,models were evaluated according to rat general state,changes in body weight and rectal temperature. Serum metabonomic profiles were detected using LC-MS technique. Difference in inter-group metabolite spectrograms was analyzedusing orthogonal partial least squares discriminant analysis（ OPLS-DA）. Potential biomarkers related to syndrome types in rat serum were selected via the parameter of variable importance in the projection（ VIP）. Results The weight of rats in the PQDS group and the PYDS group decreased more significantly after modeling. The difference in pre-post weight was statistically significant from that of the normal control group（P〈 0. 01）. It was more obviously lowered in the PYDS group than in the PQDS group（P〈 0. 05）. Compared with the normal control group,the rectal temperature of rats in the PYDS group and the PQDS group decreased（P 〈0. 05,P〈 0. 01）. It decresed more in the PYDS group than in the PQDS group（P 〈0. 05,P〈 0. 01）.Compared with the normal control group,levels of PC（ 19∶ 0） / PE（ 22∶ 0）,PC（ 17∶ 0） / PE（ 20∶ 0）,capric acid,oleic acid,stearic acid,succinic acid,fumaric acid,malic acid,g

关 键 词：液质联用 血清 脾气虚证 脾阳虚证 代谢组学 

分 类 号：R228[医药卫生—中医基础理论] R-332[医药卫生—中医学]