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基于粪便代谢组学与燥证相关性探讨岭南炮制枳壳的润燥效应

[Dryness-alleviating effect of processed Aurantii Fructus from Lingnan based on correlation of fecal metabolomics and dryness].

作者信息

Huang Ying-Ying, Yang Ting, Wang Qi-Qi, Long Jiang-Ling, Li Qin-Ru, Xu Qi-Jian, Huang Ai-Hua, Wang Jin-Yu, Xia Quan

机构信息

School of Pharmaceutical Science, Guangzhou University of Chinese Medicine Guangzhou 510006, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2023 Jan;48(1):82-95. doi: 10.19540/j.cnki.cjcmm.20220712.301.

Abstract

With the approach of untargeted metabolomics and correlation analysis, this study aimed to explore the mechanism of Aurantii Fructus from Lingnan region in alleviating dryness by analyzing the different effects of raw Aurantii Fructus(RAF) and processed Aurantii Fructus(PAF) on fecal endogenous metabolism in normal rats. Eighteen Sprague-Dawley(SD) rats were randomly divided into a control group(C), an RAF group(10 g·kg(-1)), and a PAF group(10 g·kg(-1)). After seven days of administration, the effects of RAF and PAF on dryness-related indexes were compared, including water intake, fecal water content, salivary secretion, the expression of AQP5, VIP, and 5-HT in the submandibular gland, as well as the expression of AQP3, VIP, and 5-HT in the colon. The fecal samples in each group were determined by LC-MS. Multivariate statistical analysis and Pearson correlation coefficient were used for screening the differential metabolites and metabolic pathways in alleviating dryness of RAF. The results indicated that both RAF and PAF showed certain dryness, and the dryness of RAF was more significant. Moreover, PAF could alleviate dryness of RAF to a certain extent by reducing the water intake, fecal water content, and the expression of AQP3, VIP, and 5-HT in the colon and increasing the salivary secretion and the levels of AQP5, VIP, and 5-HT in the submandibular gland. According to the analysis of fecal metabolomics, 99 and 58 metabolites related to dryness were found in RAF and PAF respectively, where 16 of them played an important role in alleviating dryness of RAF. Pathway analysis revealed that the mechanism of PAF in alleviating dryness of RAF was presumably related to the regulation of riboflavin metabolism, purine metabolism, arginine biosynthesis, pyrimidine metabolism, alanine metabolism, aspartate metabolism, glutamate metabolism, and retinol metabolism pathways. This study suggested that PAF might alleviate dryness of RAF by affecting the metabolic levels of the body, which provides a new basis for further clarifying the processing mechanism of PAF.

摘要

采用非靶向代谢组学及相关性分析方法,本研究旨在通过分析生品枳壳(RAF)和炮制品枳壳(PAF)对正常大鼠粪便内源性代谢的不同影响,探讨岭南地区枳壳润燥的作用机制。将18只Sprague-Dawley(SD)大鼠随机分为对照组(C)、RAF组(10 g·kg⁻¹)和PAF组(10 g·kg⁻¹)。给药7天后,比较RAF和PAF对润燥相关指标的影响,包括饮水量、粪便含水量、唾液分泌量、下颌下腺中AQP5、VIP和5-HT的表达,以及结肠中AQP3、VIP和5-HT的表达。采用液相色谱-质谱联用仪测定各组大鼠粪便样本。运用多元统计分析和Pearson相关系数筛选RAF润燥的差异代谢物及代谢途径。结果表明,RAF和PAF均有一定的润燥作用,且RAF的润燥作用更显著。此外,PAF可通过降低饮水量、粪便含水量以及结肠中AQP3、VIP和5-HT的表达,增加唾液分泌量以及下颌下腺中AQP5、VIP和5-HT的水平,在一定程度上缓解RAF的润燥作用。通过粪便代谢组学分析,分别在RAF和PAF中发现99种和58种与润燥相关的代谢物,其中16种在缓解RAF润燥中起重要作用。通路分析显示,PAF缓解RAF润燥的机制可能与核黄素代谢、嘌呤代谢、精氨酸生物合成、嘧啶代谢、丙氨酸代谢、天冬氨酸代谢、谷氨酸代谢和视黄醇代谢途径的调控有关。本研究提示,PAF可能通过影响机体代谢水平来缓解RAF的润燥作用,为进一步阐明PAF的炮制机制提供了新依据。

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