广藿香对贵州小型猪血清代谢组的影响及其机制探讨

doi:10.12300/j.issn.1674-5817.2022.186

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (3): 253-261.DOI: 10.12300/j.issn.1674-5817.2022.186

广藿香对贵州小型猪血清代谢组的影响及其机制探讨

陆涛峰¹(), 张慧²(), 周洁³, 李倩¹, 吴曙光¹, 吴延军¹()()

^1.贵州中医药大学实验动物研究所, 贵阳 550025
^2.广州中医药大学东莞医院, 东莞 523000
^3.上海懿尚生物科技有限公司, 上海 201319

收稿日期:2022-12-05 修回日期:2023-02-09 出版日期:2023-06-25 发布日期:2023-07-18
通讯作者: 吴延军（1987—），男，博士，讲师，研究方向：实验动物学。E-mail：601833449@qq.com。ORCID： 0000-0002-4379-8230
作者简介:陆涛峰（1982—），男，博士，副教授，研究方向：实验动物学。E-mail：taofenglu@163.com。ORCID： 0000-0001-5881-905X；
张慧（1995—），女，硕士，中医师，研究方向：中医学。E-mail：mofans318@163.com。ORCID: 0009-0001-9384-9163
基金资助:
贵州省科技计划项目“贵州香猪疫病监测体系建立及主要疫病防控技术研究”(黔科合支撑〔2020〕1Y035)

Effects of Pogostemon cablin on Serum Metabolomiceof Guizhou Miniature Pigs and It's mechanism

Taofeng LU¹(), Hui ZHANG²(), Jie ZHOU³, Qian LI¹, Shuguang WU¹, Yanjun WU¹()()

^1.Institute for Laboratory Animal Research, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China
^2.Dongguan Hospital, Guangzhou University of Traditional Chinese Medicine, Dongguan 523000, China
^3.Shanghai Yishang Biotechnology Co. , LTD. , Shanghai 201319, China

Received:2022-12-05 Revised:2023-02-09 Published:2023-06-25 Online:2023-07-18
Contact: WU Yanjun (ORCID: 0000-0002-4379-8230), E-mail: 601833449@qq.com

1. 该文献附件资料.xls(128KB)

摘要/Abstract

摘要：

目的基于液相色谱-串联质谱联用（liquid chromatography-tandem mass spectormetry，LC-MS/MS）技术研究广藿香对贵州小型猪血清代谢的影响，探讨其药理学作用机制。方法将9头贵州小型猪随机分为广藿香给药组（5头）和对照组（4头）。广藿香给药组按0.5 g·头^-1·d^-1剂量通过拌料经口饲喂广藿香中药配方颗粒，连续给药8 d；对照组正常饲喂，不额外给药。用药结束后采集各组动物的血清样品，采用LC-MS/MS技术分析各组样品的血清代谢物，对代谢组数据进行注释并与KEGG、HMDB和Lipidmaps数据库比对，通过偏最小二乘判别分析（partial least squares discriminant analysis，PLS-DA）、组间聚类、差异代谢物分析和功能富集等生物信息学分析方法，筛选差异代谢生物标志物及其可能的代谢通路。结果在广藿香给药组和对照组的血清样本中共鉴定出443个血清代谢物，筛选出44个差异显著（P<0.05）的代谢物，其中显著上调（P<0.01）的代谢物为肉桂酰甘氨酸、N-苄基-N-异丙基-N'-（4-三氟甲基）苯尿素、亚牛磺酸、D-葡萄糖-6-磷酸、顺-2-癸烯酸、二十碳二烯酸（顺-11，14）、前列腺素A2和10-羟基癸烯酸，显著下调（P<0.01）的代谢物为溶血磷脂酰胆碱22∶5、溶血磷脂酸22∶6和溶血磷脂酸22∶5。差异代谢物主要富集在丙氨酸、天冬氨酸和谷氨酸代谢通路（MapID：map00250），以及牛磺酸和亚牛磺酸代谢通路（MapID：map00430）。结论广藿香能显著影响贵州小型猪的溶血磷脂酸类物质的代谢，可能通过影响天冬氨酸-谷氨酸代谢通路、牛磺酸-亚牛磺酸代谢通路，起到缓解机体氨基酸代谢紊乱、调节炎性反应发生等功能。

关键词: 代谢组学, 广藿香, 贵州小型猪, 中药药理, 作用机制

Abstract:

Objective Based on the liquid chromatography-tandem mass spectormetry (LC-MS/MS), to study the effects of Pogostemon cablin on serum metabolism of Guizhou miniature pigs， and to explore its pharmacological mechanism. Methods Nine healthy Guizhou miniature pigs were divided into two groups, namely Pogostemon cablin drug group (n=5) and control group (n=4). The pigs in Pogostemon cablin drug group were orally fed with traditional Chinese medicine formula granules, each 0.5 g per day, for consecutive 8 days, while those in control group were given normal feeding without additional treatment. After the feeding experiment, serum samples were collected and analyzed using the LC-MS/MS technology. The metabolomics data was annotated and compared with KEGG, HMDB and Lipidmaps databases. Bioinformatics analysis methods including partial least squares discriminant analysis (PLS-DA), intergroup clustering, differential metabolite analysis and functional enrichment were used to screen differential metabolic biomarkers and their possible metabolic pathways. Results Forty-four differential metabolites (P<0.05) were screened out from the 443 metabolites, eight differential metabolites were significantly up-regulated (P<0.01), namely cinnamoylglycine, N-benzyl-N-isopropyl-N'-[4-(trifluoromethoxy) phenyl]urea, hypotaurine, D-glucose 6-phosphate, cis-2-decenoic acid, 11(Z),14(Z)-eicosadienoic acid, prostaglandin A2 and 10-hydroxydecanoic acid, and three differential metabolites were significantly down-regulated (P<0.01), namely lysophosphatidyl choline 22:5, lysophosphatidic acid 22:6 and lysophosphatidic acid 22:5. The differential metabolites were mainly enriched in the metabolic pathways of alanine, aspartate and glutamate metabolism (MapID: map00250) and taurine and hypotaurine metabolism (MapID: map00430). Conclusion Pogostemon cablin can significantly affect the metabolism of lysophosphatidic acids in porcine, and relieve the disorder of amino acids metabolism and regulate the occurrence of inflammation by affecting the metabolic pathways of alanine, aspartate and glutamate metabolism and taurine and hypotaurine metabolism.

Key words: Serum metabolomics, Pogostemon cablin, Guizhou miniature pig, Pharmacology of traditional Chinese medicine, Action mechanism

中图分类号:

R-332

陆涛峰, 张慧, 周洁, 李倩, 吴曙光, 吴延军. 广藿香对贵州小型猪血清代谢组的影响及其机制探讨[J]. 实验动物与比较医学, 2023, 43(3): 253-261.

Taofeng LU, Hui ZHANG, Jie ZHOU, Qian LI, Shuguang WU, Yanjun WU. Effects of Pogostemon cablin on Serum Metabolomiceof Guizhou Miniature Pigs and It's mechanism[J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 253-261.

图/表 5

图1 正（A）负（B）离子模式下QC样本的两两相关性分析注：QC指质量控制样本；pos指阳离子模式；neg指阴离子模式。

Figure 1 Pairings correlation of QC samples in positive （A） and negative （B） ion modeNote：QC， Quality control sample； pos， Positive ion mode； neg， Negative ion mode.

图2 广藿香给药后猪血清代谢物的LC-MS/MS总离子流图（A）及PLS-DA得分图（B）注：A，负离子模式下的LC-MS/MS总离子流图；B，代谢物的偏最小二乘判别分析（PLS-DA）得分图。GHX为广藿香给药组，CT为不加药、正常饲喂的对照组；RT，治疗时间。

Figure 2 The results of LC-MS/MS negative mode scanning total ion flow diagram （A） and PLS-DA score map （B） of serum metabolites of pigs after administration of Pogostemon cablinNote：A， LC-MS/MS negative mode scanning total ion flow diagram； B， Partial least squares discriminant analysis （PLS-DA） score map for metabolite data. GHX， Pogostemon cablin drug group given 0.5 g/head of traditional Chinese medicine formula granules per day by mixing materials through oral feeding for consecutive 8 days； CT， The control group given normal feeding without additional treatment； RT， Treatment time.

图3 广藿香给药后猪血清中差异代谢物的注释结果注：A～C分别为HMDB（Human Metabolome Database）数据库、KEGG（Kyoto Encyclopedia of Genes and Genomes）数据库和LIPID Maps数据库的对比注释结果。

Figure 3 Annotation results of serum metabolites of pigs after administration of Pogostemon cablinNote：A to C are the comparative annotation results of HMDB （Human Metabolome Database） database， KEGG （Kyoto Encyclopedia of Genes and Genomes） database， and LIPID Maps database， respectively.

图4 广藿香组（GHX）和对照组（CT）猪血清差异代谢物的火山图（A）、聚类热图（B）和相关性分析图（C）注：GHX为广藿香给药组，CT为不加药、正常饲喂的对照组；FC，差异倍数。

Figure 4 Volcanic map （A）， clustering heat map （B） and correlation analysis diagram （C） of differential serum metabolites of pigs after administration of Pogostemon cablinNote：GHX， Pogostemon cablin drug group given 0.5 g/head of traditional Chinese medicine formula granules per day by mixing materials through oral feeding for consecutive 8 days； CT， The control group given normal feeding without additional treatment； FC， Fold change.

表1 广藿香给药后影响猪血清主要差异代谢物的基本信息

Table 1 The basic information of the main differential metabolites of pigs after administration of Pogostemon cablin

代谢物 Metabolites	代谢物中文名称 Metabolites in Chinese	保留时间/min Retention time/min	log₂(FC)	P值 P value
Cinnamoylglycine	肉桂酰甘氨酸	8.378	3.670	0.001
N-benzyl-N-isopropyl-N'-[4-(trifluoromethoxy)phenyl] urea	N-苄基-N-异丙基-N'-(4-三氟甲基)苯尿素	4.541	2.217	0.005
Hypotaurine	亚牛磺酸	14.485	1.703	0.008
D-Glucose 6-phosphate	D-葡萄糖-6-磷酸	1.158	1.595	0.003
cis-2-Decenoic acid	顺-2-癸烯酸	12.057	1.536	0.002
11(Z),14(Z)-Eicosadienoic acid	二十碳二烯酸(顺-11,14)	14.820	1.349	0.007
Prostaglandin A2	前列腺素A2	12.727	1.289	0.008
10-Hydroxydecanoic acid	10-羟基癸烯酸	11.707	1.261	0.004
Lysophosphatidyl choline 22:5	溶血磷脂酰胆碱22:5	14.673	-2.190	0.002
Lysophosphatidyl acid 22:6	溶血磷脂酸22:6	14.010	-2.201	<0.001
Lysophosphatidyl acid 22:5	溶血磷脂酸22:5	14.139	-2.361	0.003

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Effects of Pogostemon cablin on Serum Metabolomiceof Guizhou Miniature Pigs and It's mechanism

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