摘要
该研究采用代谢组学方法分析广陈皮对脾虚痰湿证内源性代谢产物的影响,并探究广陈皮治疗脾虚痰湿证的相关作用机制。采用多因素造模法建立脾虚痰湿证大鼠模型。通过观察大鼠肝组织病理变化,并测定血浆中甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)等病理生化指标含量,初步评价广陈皮对脾虚痰湿证模型大鼠的干预作用。采用免疫组化技术检测大鼠肾AQP2、结肠AQP3、颌下腺AQP5的表达,评估陈皮对脾虚痰湿证大鼠的水通道蛋白表达作用。进一步采用UHPLC-ESI-MS/MS技术对大鼠血浆样本进行代谢轮廓分析,应用主成分分析(PCA)、正交偏最小二乘-判别分析(OPLS-DA)等多种方法进行模式识别,结合t检验和变量投影重要性(VIP)筛选差异代谢物,并基于MetaboAna-lyst 5.0进行通路分析。实验表明广陈皮可改善脾虚痰湿证大鼠肝组织病理变化,升高大鼠血浆中HDL-C水平,并降低TG、TC、LDL-C水平,增强对脾虚痰湿证大鼠肾AQP2、结肠AQP3以及颌下腺AQP5的表达。此外,UHPLC-ESI-MS/MS共筛选出脾虚痰湿证差异代谢物87个(其中39个代谢物水平明显上调升高,48个代谢物水平显著下降),差异最大的代谢物依次为甘氨酸、L-异亮氨酸、N-乙酰-L-酪氨酸、黄嘌呤、次黄嘌呤、葫芦巴碱等。差异代谢物主要富集于类固醇激素生物合成、亚油酸代谢、嘌呤代谢等通路。该研究采用代谢组学方法能区分并揭示脾虚痰湿证的特征性代谢模式,OPLS-DA判别模型的初步构建为脾虚痰湿证的中医证候辨识提供了客观辨证依据,为从小分子层面及整体水平探究中医证候的生物学基础提供思路与方法。
The present study analyzed the effect of Citri Reticulatae Pericarpium on endogenous metabolites in spleen deficiency and phlegm dampness syndrome by metabolomics, and explored the underlying mechanism of Citri Reticulatae Pericarpium in the treatment of spleen deficiency and phlegm dampness syndrome.The model of spleen deficiency and phlegm dampness syndrome was induced in rats by the multi-factor modeling method.The intervention effects of Citri Reticulatae Pericarpium on rats with spleen deficiency and phlegm dampness syndrome were preliminarily evaluated by observing the pathological changes of rat liver tissues and measuring the plasma content of pathological and biochemical indexes such as triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C).Immunohistochemistry was used to detect the expression of AQP2 in the kidney, AQP3 in the colon, and AQP5 in the submandibular gland, and the effect of Citri Reticulatae Pericarpium on aquaporin expression in rats with spleen deficiency and phlegm dampness syndrome was evaluated.Furthermore, UHPLC-ESI-MS/MS was used to analyze the metabolic profiles of rat plasma samples.Multiple methods, such as principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used for pattern recognition.Differential metabolites were screened out by t-test and variable importance in projection(VIP), followed by pathway analysis based on MetaboAnalyst 5.0.As revealed by experimental results, Citri Reticulatae Pericarpium could improve the pathological changes of liver tissues, increase the levels of HDL-C in the plasma, reduce the levels of TC, TG, and LDL-C, and enhance the expression of AQP2 in the kidney, AQP3 in the colon, and AQP5 in the submandibular gland of rats with spleen deficiency and phlegm dampness syndrome.In addition, 87 differential metabolites of spleen deficiency and phlegm dampness syndrome were screened out by UHPLC-ESI-MS/MS(the levels of 39 met
作者
吴蓓
申梦园
陈鸿平
刘友平
王福
陈林
WU Bei;SHEN Meng-yuan;CHEN Hong-ping;LIU You-ping;WANG Fu;CHEN Lin(State Key Laboratory of Southuestern Chinese Medicine Resources,School of Pharmacy,Chengdu Universiy of Traditional Chinese Medicine,Chengdu 611137,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第15期4136-4147,共12页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(8197142602)。
关键词
广陈皮
脾虚痰湿证
代谢组学
差异代谢物
UHPLC-ESI-MS/MS
Citri Reticulatae Pericarpium
spleen deficiency and phlegm dampness syndrome
metabolomics
differential metabolites
UHPLC-ESI-MS/MS
