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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、棕榈酰肉碱、肉豆蔻酰肉碱、乙酰左旋肉碱及亚油肉碱相对含量较脾气虚组显著降低。结论脾气虚及脾阳虚模型大鼠血清中多种代谢物的相对含量发生了显著变化,并初步获得了一些可能与脾虚证候相关的潜在小分子生物标志物。这可能为探索脾虚的中医证候的科学内涵、病理机制提供了一定的数据参考。
Objective To investigate the changes of serum metabolites in spleen-deficiency syndrome and spleen-yang deficiency syndrome by using liquid chromatography-mass spectrometry (LC-MS), and to explore the essence of syndrome of spleen deficiency syndrome from the level of small molecule metabolites. Methods Twenty-one SPF male Sprague-Dawley rats were randomly divided into normal group, spleen-qi deficiency group and spleen-Yang deficiency group, with 7 rats in each group. Spleen qi deficiency syndrome modeling method using the first satiation for 1 day, then fasting 2 days, free access to water, daily swimming to exhaustion, water temperature 35 ~ 37 ℃, for 15 days. Spleen deficiency syndrome modeling method using spleen qi deficiency syndrome plus irrigation service 20% senna flooding agent (2 m L / 100 g), morning and evening 1, for 1 week. At the end of the modeling, the model was evaluated according to the general situation of rats, body weight changes and rectal temperature changes. Serum metabolomics was detected by LC-MS. The results of orthogonal partial least squares discriminant analysis Metabolomics spectrum differences were also investigated, and the syndrome-related biomarkers in serum were selected by variable importance projection (OPLS-DA). Results Compared with the model group before the model, the body weight of the spleen deficiency group and the spleen yang deficiency group were significantly lower than that of the normal group (P <0.01). The difference of body weight between the two groups was statistically significant (P <0.01) (P <0.05). Compared with the normal group, the rectal temperature decreased in the spleen deficiency group and the spleen yang deficiency group, while the spleen deficiency group decreased more than the spleen deficiency group (P <0.05, P <0.01). Compared with normal group, PC (19:0) / PE (22:0), PC (17:0) / PE (20:0), capric acid, oleic acid, stearic acid , Fumaric acid, fumaric acid, malic acid and glucose. The contents of arachidonic acid, linolenic acid, dodecenoic acid, androsterone, 4-heptanone, DHAP (6:0) and uridine decreased. Compared with the spleen deficiency group, the relative contents of PC (22:1), PC (22:6), PE (18:0) / PC (15:0), retinol and deoxycytidine were significantly increased (19: 3), PC (20: 3), PC (17: 0) / PE PC (19: 0) / PE (22: 0) PC: 17: 1 PE: 20: 1 PC: 16: 1 PE: 19: 1 Cholic acid, hippuric acid, , Undecanedicarboxylic acid, palmitoleic acid, hydroxystearic acid, eicosatrienoic acid, phenylalanine, tyrosine, glutamic acid, serine, carbamoyl aspartic acid, palmitoyl carnitine , Myristoyl carnitine, acetyl-L-carnitine and linoleic acid relative content than Spleen deficiency group was significantly lower. Conclusion The relative contents of various metabolites in serum of spleen-qi deficiency and spleen-yang deficiency rats have changed significantly, and some potential small molecular biomarkers that may be related to the syndrome of spleen deficiency were preliminarily obtained. This may provide some data reference for exploring the scientific connotation and pathological mechanism of TCM syndrome of spleen deficiency.