Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (2): 368-378.doi: 10.3864/j.issn.0578-1752.2023.02.013

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles     Next Articles

Evaluation of Regulatory Effect of Guiqi Yimu Oral Liquid on Rumen of Postpartum Dairy Cows Based on UPLC-MS/MS Metabolomics Technology

TANG YuLin1(),ZHANG Bo1,REN Man1,ZHANG RuiXue1,QIN JunJie2,ZHU Hao2,GUO YanSheng1()   

  1. 1College of Agriculture, Ningxia University, Yinchuan 750021
    2Beijing Center Biology Co., Ltd, Beijing 102600
  • Received:2021-09-22 Accepted:2022-08-22 Online:2023-01-16 Published:2023-02-07

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Abstract:

【Background】Under the background of ban on antibiotics in animal feed at the current, the use of low residue, non-toxic Chinese herbal medicine preparations for perinatal health care has been favored by the majority of breeding enterprises. The previous studies found that Guiqi Yimu oral liquid could reduce inflammatory response, enhance immunity and relieve negative energy balance in postpartum cows, but the detailed mechanism of action was unclear. 【Objective】This study aimed to reveal the mechanism of action of Guiqi Yimu oral liquid from a new perspective through the regulatory effect of the prescription on rumen metabolism in postpartum cow with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) metabolomics technology. 【Method】 18 healthy parturient Holstein cows with similar body condition and 2-3 parities were selected and averagely divided into the control group and the test group. From Day 0 to Day 6 after birth, the cows of test group were administrated Guiqi Yimu oral liquid, and those of control group were orally administrated water oral. Rumen fluids of all cows were collected at postpartum Day 0 before administration and postpartum Day 7 before first meal. After pretreatment, the metabolites in rumen were qualified and quantified with UPLC-MS/MS technology. The obtained data was imported into MetaboAnanlyst 5.0 software to perform principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA), investigating alteration in metabolic profiles of rumen fluids between the two groups, and screening differential metabolites for cluster and metabolic pathway enrichment analysis. 【Result】 The rumen metabolic profile of the test group was obvious different with the control group at postpartum Day 7 due to the concentrations of 43 rumen metabolites changed significantly (VIP>1, P<0.05). Compared wtih the control group, 8 metabolites were up-regulated (P<0.05), including 9-oxoODE, punicic acid, 6-dimethylaminopurine, L-proline, maleic acid, N-acetyl-5-hydroxytryptamine, pantothenol, and formononetin, and 35 metabolites were down-regulated in the test group (P<0.05), including organic acids and their derivatives, nucleotides and their metabolites, carbohydrates and their metabolites. Metabolic pathway enrichment analysis showed Guijiyimu oral liquid obviously altered rumen histidine metabolism, pentose and glucuronic acid mutual conversion, arginine and proline metabolism tryptophan metabolism, pyrimidine metabolism and purine metabolism pathways in postpartum dairy cows. 【Conclusion】 Guiqi Yimu oral liquid promoted the metabolic function of rumen flora in postpartum dairy cows, which was one of the mechanisms to alleviate the negative energy balance, enhance immunity, anti-inflammation and maintain intestinal homeostasis in postpartum dairy cows.

Key words: Guiqi Yimu oral liquid, postpartum dairy cows, rumen fluid, UPLC-MS/MS metabolomics

Fig. 1

Effects of Guiqi Yimu Oral Liquid on PCA in rumen fluid of dairy cows on day 0 postpartum a: A compared with F, b: B compared with G"

Fig. 2

2D scatter plot (a) and validation map (b) of OPLS-DA metabolites in rumen fluid of dairy cows at 7 d postpartum"

Table 1

Up-regulation of differential metabolites in rumen fluid of dairy cows at 7 days postpartum"

差异代谢物
Differential metabolite		 含量变化
Content change		 重要变量投影值Variable importance projection value P值
P value
9-氧代-10E，12Z-十八碳二烯酸
9-oxoODE[9-oxo-10E,12Z-octadecadienoic acid]		 上调Up 2.51 5.71×10-5
石榴酸 Punicic Acid 上调Up 2.11 2.09×10-3
6-二甲基氨基嘌呤 6-Dimethylaminopurine 上调Up 2.10 2.24×10-3
L-脯氨酸 L-Proline 上调Up 1.88 7.83×10-3
马来酸 Maleic Acid 上调Up 1.87 8.30×10-3
N-乙酰基5-羟色胺
O-N-Acetyl-5-Hydroxytryptamine		 上调Up 1.75 0.015
DL-泛酰醇 Pantothenol 上调Up 1.67 0.022
刺芒柄花素 Formononetin 上调Up 1.61 0.029

Table 2

Differential metabolites down-regulated in rumen fluid of dairy cows at 7 d postpartum"

差异代谢物
Differential metabolite		 含量变化
Content change		 重要变量投影值
Variable importance projection value		 P值
P value
1-辛烯-3-酮 1-Octen-3-one 下调Down 2.54 3.64×10-5
1,2-二氯乙烷 1,2-Dichloroethane 下调Down 2.53 4.16×10-5
冰片 (+)-borneol 下调Down 2.52 4.81×10-5
6-甲基-5-庚烯-2-酮 6-Methyl-5-hepten-2-one 下调Down 2.51 5.23×10-5
(±)-萜烯-4-醇 p-Menth-1-en-4-ol 下调Down 2.50 6.17×10-5
磷酸三乙酯 Triethyl phosphate 下调Down 2.48 7.72×10-5
（±）9,10-二羟基-12Z-十八碳烯酸
9,10-DiHOME [(+-)9,10-dihydroxy-12Z-octadecenoic acid]		 下调Down 2.29 4.91×10-4
溶血卵磷脂酰胆碱16:0 Lysopc 16:0 下调Down 1.95 5.32×10-3
2,6-二氨基庚二酸 2,6-Diaminooimelic Acid 下调Down 1.91 6.85×10-3
吲哚-3-丙酸 3-Indolepropionic Acid 下调Down 1.89 7.40×10-3
溶血磷脂酰胆碱 18:3 Lysopc 18:3 下调Down 1.88 7.95×10-3
3-羟基-2-吡啶甲酸 3-Hydroxypicolinic acid 下调Down 1.88 8.04×10-3
3-羟基 - DL-犬尿氨酸 4-3-Hydroxy-DL-kynurenine 下调Down 1.87 8.15×10-3
脱氧鸟苷 Deoxyguanosine 下调Down 1.85 9.18×10-3
胸腺嘧啶 Thymine 下调Down 1.78 1.30×10-2
猪去氧胆酸 Hyodeoxycholic Acid 下调Down 1.76 0.014
2，2'-硫代二乙醇酸 Thiodiglycolic Acid 下调Down 1.71 0.018
尿刊酸 Urocanic Acid 下调Down 1.54 0.019
对苯二甲酸 Phthalic Acid 下调Down 1.70 0.019
L-胱氨酸 L-Cystine 下调Down 1.69 0.019
2,4-二羟基苯甲酸 2,4-Dihydroxybenzoic Acid 下调Down 1.67 0.021
溶血卵磷脂酰乙醇胺16:0 Lysope 16:0 下调Down 1.67 0.022
新蝶呤 Neopterin 下调Down 1.63 0.025
腐胺 Putrescine 下调Down 1.60 0.029
2-吲哚甲酸 Indole-2-Carboxylic Acid 下调Down 1.59 0.031
蛋氨酸亚砜 Methionine Sulfoxide 下调Down 1.58 0.032
乙基丙二酸Ethylmalonate 下调Down 1.57 0.032
土木香内酯 Alantolactone 下调Down 1.56 0.034
溶血卵磷脂酰胆碱18:2 Lysopc 18:2 下调Down 1.55 0.034
十八碳烯酸(反-9) Elaidic Acid (C18:1N9T) 下调Down 1.56 0.034
木糖 Xylose 下调Down 1.55 0.035
尿苷 Uridine 下调Down 1.55 0.036
9S-羟基-10E，12Z，15Z-十八碳三烯酸
9-HOTrE [9S-hydroxy-10E,12Z,15Z-octadecatrienoic acid]		 下调Down 1.50 0.044
苯乙酰甘氨酸 N-Phenylacetylglycine 下调Down 1.49 0.045
鸟苷 Guanosine 下调Down 1.48 0.046

Fig. 3

Clustering heat map of differential metabolites in the first 10 rumen fluids of dairy cows at 7 days postpartum"

Fig. 4

Metabolic pathways and partial pathway names of differential metabolites in rumen juice of dairy cows at 7 days postpartum 1: Histidine metabolism; 2: Interconversion of pentose and glucuronic acid; 3: Arginine and proline metabolism; 4: Tryptophan metabolism; 5: Pyrimidine metabolism; 6: Purine metabolism"

Fig. 5

Violin diagram of differential metabolites in metabolic pathways between the control group and the test group 7 days postpartum"

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