基于“运脾生血”理论的熟地黄补血作用代谢组学研究  

Metabolomic profiling reveals blood-tonifying effect of Rehmanniae Radix Praeparata based on theory of activating spleen and generating blood

作　　者：王洋洋[1] 冯卫生[2] 王秋红 匡海学[1] WANG Yang-yang;FENG Wei-sheng;WANG Qiu-hong;KUANG Hai-xue(Key Laboratory of Basic and Application Research of Beiyao,Ministry of Education,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Henan University of Chinese Medicine,Zhengzhou 450046,China;School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]黑龙江中医药大学教育部北药基础与应用研究重点实验室,黑龙江哈尔滨150040 [2]河南中医药大学,河南郑州450046 [3]广东药科大学中药学院,广东广州510006

出　　处：《中国中药杂志》2022年第13期3562-3568,共7页China Journal of Chinese Materia Medica

基　　金：黑龙江省“头雁”团队支持项目(黑龙江省头雁行动领导小组文件[2019]5号);国家中医药管理局中医药传承与创新“百千万”人才工程——岐黄工程首席科学家支持项目(国中医药人教函[2021]7号);国家中医药管理局中医药传承与创新“百千万”人才工程(岐黄工程)岐黄学者支持项目(国中医药人教函[2018]284号);国家重点研发计划项目(2017YFC1702800);2022年全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号);第七批全国老中医药专家学术经验继承工作项目(国中医药人教函[2022]76号)。

摘　　要：该研究基于“运脾生血”理论探讨熟地黄对血虚证大鼠脾脏代谢组学的研究。采用环磷酰胺结合N-乙酰苯肼的方法建立血虚证模型,并采用UPLC-Q-TOF-MS技术对血虚模型的脾脏样本进行代谢组学分析,对脾脏样本进行主成分分析(PCA)、正交偏二乘判别分析(OPLS-DA),对潜在生物标志物的相对定量结果进行MEV热图聚类分析以及体内代谢网络的构建,结合血虚证大鼠血常规指标,共同阐释熟地黄基于“运脾生血”理论的补血作用机制。结果表明,血虚证模型组白细胞数(WBC)、红细胞数(RBC)、血小板(PLT)和红细胞压积(HCT)等血常规指标显著降低,说明血虚证模型复制成功;在给予熟地黄后血虚证模型大鼠血常规指标显著改善,说明熟地黄对血虚证大鼠发挥了明显的补血作用。同时在代谢组学分析中,共鉴定了41个潜在生物标志物。PCA结果以及潜在生物标志物聚类分析中熟地黄对血虚证模型大鼠脾脏样本的整体代谢轮廓也均表现出明显改善作用。这些潜在生物标志物主要涉及机体的三羧酸循环(tricarboxylic acid,TCA)、烟酸和烟酰胺代谢,苯丙氨酸代谢,酪氨酸代谢,牛磺酸和亚牛磺酸代谢,鞘脂代谢。基于“运脾生血”理论以及血虚证大鼠模型推测熟地黄可能主要通过调节脾脏相关的能量代谢、外周血生成以及血细胞氧化损伤等相关代谢通路发挥其补血的作用。Based on the theory of activating spleen and generating blood,this study explored the effect of Rehmanniae Radix Prae-parata on the spleen metabolome of the rat model with blood deficiency syndrome.The rat model of blood deficiency syndrome was established by combining with cyclophosphamide(CTX)and N-acetyl-phenylhydrazine(APH),and the metabolomes of the spleen samples were analyzed with ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).Principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were carried out for the metabolite profiles of spleen samples.The MEV heatmap and metabolic network were established based on the potential biomarkers.Finally,the blood routine indexes were combined with the metabolomic profile to reveal the mechanism of Rehmanniae Radix Praeparata in activating spleen and generating blood.The treatment with CTX and APH decreased the blood routine indexes such as white blood cell count(WBC),red blood cell count(RBC),platelet(PLT),and hematocrit(HCT),indicating that the rat model of blood deficiency syndrome was successfully established.The administration of Rehmanniae Radix Praeparata significantly improved the blood routine indexes,which suggested that Rehmanniae Radix Praeparata played a role in replenishing blood.In addition,the metabolomics analysis identified 41 potential biomarkers.The PCA and MEV heatmap also showed significant improvement effect of Rehmanniae Radix Praeparata on the spleen metabolic profile.These potential biomarkers were mainly involved in tricarboxylic acid cycle,niacin and nicotinamide metabolism,phenylalanine metabolism,tyrosine metabolism,taurine and hypotaurine metabolism,and sphingolipid metabolism.Therefore,we hypothesize that Rehmanniae Radix Praeparata may regulate energy metabolism,peripheral blood production,and oxidative injury of hemocytes to tonify blood.

关 键 词：熟地黄 “运脾生血”理论 血虚证 脾脏代谢组学 

分 类 号：R285.5[医药卫生—中药学]