Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 10: 7074 - 7086

DOI: https://doi.org/10.12418/CJAN2025.576

RESEARCH PAPER

Effects of Houttuynia cordata on Milk Antioxidant Capacity and Shelf Life based Metabolomics

  • FENG Lunjie , 1 ,
  • TANG Huayu 1 ,
  • WU Tianci 1 ,
  • CHEN Ling 2 ,
  • CHEN Shuai 3 ,
  • CHEN Liwen 3 ,
  • JIANG Chengquan 2 ,
  • YAN Honglin , 1, *
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  • 1 College of Life Science and Agri-Forestry, Southwest University of Science and Technology, Mianyang 621000, China
  • 2 Sichuan College of Traditional Chinese Medicine, Mianyang 621000, China
  • 3 Mianyang Anzhou District Hongfeng Dairy Cattle Farming Co., Ltd., Mianyang 621000, China
*associate professor, E-mail:

Received date: 2025-01-03

  Online published: 2025-10-15

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Abstract

This experiment was conducted to study the effects of Houttuynia cordata on milk antioxidant capacity and shelf life based metabolomics. Twenty healthy Holstein cows with similar body weight were divided into 2 groups according to the principle of similar parity, lactation days and milk yield, with 10 replicates in each group and 1 cow in each replicate. The control group was fed a basal diet without Houttuynia cordata, and the experimental group was fed the experimental diet which used 2% Houttuynia cordata replacing an equivalent amount of corn silage in the basal diet. The experiment lasted for 5 weeks. At the end of the experiment, the milk samples were collected to measure the bioactive substances contents and antioxidant indices, and the metabolomics techniques were employed for small molecule differential metabolite analysis. The fresh milk samples underwent pasteurization or boiling were subjected to a 6-day storage trial, to determine the antioxidant indices at 3 and 6 days of storage. The results showed as follows: 1) compared with the control group, the total phenolics, total flavonoids and glutathione contents and total superoxide dismutase activity of the experimental group were significantly increased (P<0.05). 2) There were 3 102 differential metabolites in milk between control group and experimental group, among them, 1 131 metabolites were significantly down-regulated (P<0.05), and 1 931 metabolites were significantly up-regulated (P<0.05). Compared with the control group, the contents of N-acetylgalactosamine and N-acetyl-D-glucosamine in the amino sugar and nucleotide sugar metabolism pathway of the experimental group were significant increased (P<0.05), and the contents of fucose-1-phosphate, D-mannose-1-phosphate, mannose-6-phosphate and N-acety-l-glucosamine-1-phosphate were significantly decreased (P<0.05); the contents of mannose-6-phosphate, fucose-1-phosphate and D-mannose-1-phosphate in the fructose and mannose metabolism pathway were significantly decreased (P<0.05); the contents of D-arabitol and D-xylitol in the pentose and glucuronate interconversion pathway were significantly increased (P<0.05); the content of L-tyrosine in the phenylalanine, tyrosine and tryptophan biosynthesis pathway was significantly increased (P<0.05); the content of riboflavin in the riboflavin metabolism pathway was significantly increased (P<0.05). 3) Compared with the control group, the glutathione content of boiled milk storage for 3 and 6 days and the total antioxidant capacity of boiled milk storage for 6 days of the experimental group were significant increased (P<0.05), and the malondialdehyde content of boiled milk storage for 3 and 6 days were significant decreased (P<0.05); the total antioxidant capacity of pasteurized milk storage for 3 and 6 days were significant increased (P<0.05), and the malondialdehyde content of pasteurized milk storage for 3 and 6 days were significant decreased (P<0.05). In conclusion, adding Houttuynia cordata in cows, diet enhance the antioxidant capacity of fresh milk, pasteurized milk and boiled milk, beneficial for extending the shelf life of milk, it may be related to the changes in composition of metabolites in milk.

Key words： Houttuynia cordata; milk; heat treatment; shelf life; antioxidant; metabolites

Cite this article

FENG Lunjie , TANG Huayu , WU Tianci , CHEN Ling , CHEN Shuai , CHEN Liwen , JIANG Chengquan , YAN Honglin . Effects of Houttuynia cordata on Milk Antioxidant Capacity and Shelf Life based Metabolomics[J]. Chinese Journal of Animal Nutrition, 2025 , 37(10) : 7074 -7086 . DOI: 10.12418/CJAN2025.576

牛奶是人类获取优质蛋白质、矿物质、碳水化合物、脂肪、维生素的重要来源,其含有的营养物质有利于维持和促进机体健康,牛奶蛋白质是生物活性肽的重要来源,牛奶中所含的蛋白质、钙、磷和其他微量元素可以促进骨骼、肌肉和神经系统的发育[1]。随着人们生活方式的改变和代谢疾病发病率的增加,消费者对含有天然抗氧化剂的食品的需求正在快速上升,牛奶及其制品消费持续向好,一些乳制品如奶油、黄油、酸奶和奶酪同样受到市场青睐。然而牛奶中的有机物质性质不稳定,牛奶的质量取决于脂肪和蛋白质含量、微生物和体细胞数量及储存时间,温度是微生物生长的所必需的环境因素,不同微生物适宜生长温度不同[2]。微生物的繁衍导致牛奶易在储藏过程中发生氧化变质,通常鲜奶在高温下的保质期从2~3 h到24 h不等[3]。影响牛奶货架期的因素有奶品质、微过滤、罐装条件、温度、包装方法和热处理等,其中热处理是生产上抑制微生物生长和繁衍最常用的方法,可以影响牛奶风味和减少乳蛋白产生,热处理越强烈,发生的变化就越彻底,是保证乳产品质量、延长产品保质期的重要工序,常见的热处理方法主要是巴氏杀菌和煮沸[4]。乳制品行业普遍采用高温短时间巴氏杀菌的工艺,在72 ℃热处理至少15 s或采用低温或保温工艺在63~66 ℃下加热至少30 min[5]。牛奶中细菌种类和数量直接影响其品质,鱼腥草的主要抑菌活性成分癸乙醛对多种细菌、抗酸杆菌和真菌均有明显的抑菌作用[6]。牛奶中嗜冷菌是评估原料奶细菌质量的生物学指标,其数量超过1.3×105 CFU/mL则在冷藏或运输过程中易导致原料奶出现质量缺陷[7]。牛奶中嗜冷菌数量过高会产生较多具有热稳定性质的蛋白酶和脂肪酶,而热处理并不能完全将其分解,影响牛奶货架期,造成产品质量缺陷;此外,大多数嗜冷细菌可以在牛奶储存和加工设备上形成生物膜,这些生物膜具有生物转移潜力,是微生物污染的持续来源[8]。研究发现,具有抗氧化作用的草药和香料可以有效延长乳制品的保质期,且副作用小[9]。
鱼腥草作为一种药食同源的中草药,在中国有悠久的使用历史,其富含生物碱、精油、多酚和类黄酮等多种生物活性物质,具有抗氧化、抗炎和抗菌作用,是一种可以替代抗生素的绿色饲料添加剂[10]。鱼腥草中的多酚和黄酮类生物活性物质具有抗氧化作用。大鼠采食添加了鱼腥草的饲粮后,能够改善硫酸庆大霉素诱导的氧化应激大鼠的肾脏功能,改善与肾脏损伤相关的生化和氧化指标[11]。此外,动物采食中草药会引起机体内代谢物发生改变,这些代谢物具有较好的生物学活性和药用价值,代谢组学在中草药的鉴定和分析中的应用,有助于发现潜在的生物标志物,更好的解释中草药的作用机制。综上所述,鱼腥草和热处理都有抑菌和抗氧化的作用,鱼腥草的药理作用能弥补热处理不能完全分解嗜冷菌的缺陷。通过分析代谢物的变化,可以更好地理解机体的生物化学过程和代谢途径。因此,本试验旨在研究泌乳期荷斯坦奶牛饲喂鱼腥草对鲜奶和不同热处理牛奶抗氧化能力和货架期及牛奶中代谢物的影响,以期为鱼腥草在奶牛生产中的应用提供一定的理论参考。

1 材料与方法

1.1 试验设计

本试验方案经西南科技大学动物伦理委员会批准(批准号:L2023006)。选用20头健康状态良好、胎次为(2.15±0.75)胎、泌乳天数为(135±9) d、产奶量为(26.80±1.40) kg的荷斯坦奶牛,随机分为2组,每组10个重复,每个重复1头奶牛。对照组饲喂不含鱼腥草的基础饲粮,试验组饲喂用2%鱼腥草等量代替基础饲粮中青贮玉米的试验饲粮。试验期为5周。在试验的第35天,采集牛奶样品用于活性物质、抗氧化能力以及小分子差异代谢物分析,并对牛奶样品进行为期6 d储藏试验。试验饲粮组成及营养水平见表1。
表1 试验饲粮组成及营养水平(干物质基础)

Table 1 Composition and nutrient levels of experimental diets (DM basis) %

项目
Items		 基础饲粮
Basal
diet		 试验饲粮
Experimental
diet
原料Ingredients
玉米Corn 20.30 20.30
豆粕Soybean meal 7.40 7.40
燕麦Oats 3.70 3.70
苜蓿草Alfalfa hay 8.70 8.70
青贮玉米Corn silage 47.10 45.10
鱼腥草Houttuynia cordata 2.00
甜菜粕Sugar beet pulp 4.50 4.50
全棉籽Whole cottonseed 4.50 4.50
葡萄糖Glucose 0.40 0.40
脂肪粉Fat powder 0.20 0.20
碳酸氢钠NaHCO3 0.70 0.70
磷酸氢钙CaHPO4 0.50 0.50
氯化钠NaCl 0.50 0.50
预混料Premix1) 1.50 1.50
合计Total 100.00 100.00
营养水平Nutrient levels2)
粗蛋白质CP 15.73 15.52
中性洗涤纤维NDF 30.00 30.30
酸性洗涤纤维ADF 18.00 18.40
钙Ca 0.88 0.90
磷P 0.38 0.36
泌乳净能NEL/(MJ/kg) 6.71 6.73
总酚Total phenolics/(g/kg) 6.26 7.09
总黄酮Total flavonoids/(g/kg) 5.26 5.60

1)每千克预混料含有 Each kilogram of the premix contained the following:VA 200 000 IU,VD 40 000 IU,VE 5 000 IU,Mg 95 200 mg,Zn 1 000 mg,Co 50 mg。2)泌乳净能参照NRC(2001)计算获得,其余为实测值。NEL was a calculated value according to NRC (2001), while the others were measured values.

1.2 试验材料

本试验所用鱼腥草营养成分含量(干物质基础):粗蛋白质13.26%,粗灰分12.60%,粗脂肪43.00%,粗纤维30.60%,中性洗涤纤维42.90%,酸性洗涤纤维32.90%,钙1.06%,总磷0.23%,总能17.43 MJ/kg。

1.3 试养管理

试验期间奶牛饲养于同一圈舍中,自由采食和饮水,每日饲喂3次(06:30、13:30、20:30),挤奶3次(05:30、12:30、19:30)。奶牛的其他饲养管理与环境条件一致,每隔7 d对圈舍进行1次消毒。

1.4 样品采集

1.4.1 鱼腥草和饲粮样品

在试验开始当天,采集基础饲粮、试验饲粮和鱼腥草各500 g,置于-20 ℃冰箱保存,用于营养水平的测定。

1.4.2 牛奶样品

于试验第35天,每个重复选择1头奶牛,使用自动并列式挤奶机,采集当天05:30、12:30和19:30的牛奶,按4∶3∶3[12]的比例混合为240 mL奶样,用无菌注射器吸取3 mL装入2个1.5 mL无菌冻存管中,置于-80 ℃冰箱保存,用于牛奶非靶向代谢组学分析和抗氧化指标测定。将剩余牛奶等量分为2份,分别置于65 ℃加热30 min和煮沸1 min,分别代表巴氏杀菌牛奶和煮沸牛奶,待冷却后装入100 mL聚对苯二甲酸乙二醇酯(polyethylene terephthalate,PET)瓶中,置于4 ℃冰箱冷藏6 d,分别在冷藏3和6 d时从每个重复中取2 mL牛奶,用于抗氧化指标测定。

1.5 测定指标及方法

1.5.1 营养水平

鱼腥草和饲粮的粗蛋白质、粗脂肪、粗纤维、中性洗涤纤维、酸性洗涤纤维、粗灰分、磷、钙含量的测定分别参照GB/T 6432—2018、GB/T 6433—2006、GB/T 6434—2006、GB/T 20806—2006、NY/T 1459—2007、GB/T 6438—2007、GB/T 6437—2018和GB/T 6436—2002中方法;鱼腥草的总能按照国际标准ISO 9831:1998推荐的方法,使用氧弹式测热仪(C2000,IKA,德国)测定。

1.5.2 牛奶中总黄酮和总酚含量以及抗氧化指标

采用苏州格锐思生物科技有限公司提供的试剂盒测定牛奶中总黄酮、总酚及1,1-二苯基-2-三硝基苯肼(DPPH)含量,采用南京建成生物工程研究所提供的试剂盒测定总超氧化物歧化酶(T-SOD)活性、总抗氧化能力(T-AOC)及谷胱甘肽(GSH)、丙二醛(MDA)含量。

1.5.3 牛奶代谢组学分析

采用液相色谱-串联质谱(LC-MS/MS)法进行牛奶非靶向代谢组学的分析,由上海百趣生物医学科技有限公司完成。代谢组样品首先经过前处理去除蛋白质及杂质并提取代谢物,然后在液相色谱-质谱(LC-MS)正、负模式下分别上机检测采集信息,后续的数据分析过程中将2组数据合并分析,得到代谢物的质谱(MS)和串联质谱(MS/MS)信息。采用Progenesis QI(Waters Corporation,美国)软件进行代谢物注释和数据预处理等,最终得到代谢物列表及数据矩阵,结合t检验和正交偏最小二乘方判别分析(OPLS-DA)的变量投影重要性(VIP),以VIP>1、P<0.05为标准筛选出差异代谢物。根据所获得的代谢物通过京都基因与基因组百科全书(KEGG)进行注释和代谢物途径分析。

1.6 数据统计与分析

数据采用Excel 2019进行初步处理,用SPSS 22.0统计软件进行独立样本t检验显著性分析,试验结果以“平均值±标准误”表示,P<0.05表示差异显著。

2 结果与分析

2.1 鱼腥草对牛奶中总酚和总黄酮含量的影响

由表2可知,与对照组相比,试验组的牛奶中总酚和总黄酮含量显著增加(P<0.05)。
表2 鱼腥草对牛奶中总酚和总黄酮含量的影响

Table 2 Effects of Houttuynia cordata on contents of total phenolics and total flavonoids in milk

项目Items 对照组Control group 试验组Experimental group P值P-value
总酚Total phenolics/(μg/mL) 64.63±1.01b 75.90±0.92a <0.001
总黄酮Total flavonoids/(g/kg) 20.11±1.02b 30.09±1.13a <0.001

同行数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。下表同。

In the same row, values with different small letter superscripts mean significant difference (P<0.05), while with the same or no letter superscripts mean no significant difference (P>0.05). The same as below.

2.2 鱼腥草对牛奶中抗氧化指标的影响

由表3可知,与对照组相比,试验组的牛奶中T-SOD活性和GSH含量显著增加(P<0.05),牛奶中T-AOC及DPPH、MDA含量没有显著差异(P>0.05)。
表3 鱼腥草对牛奶中抗氧化指标的影响

Table 3 Effects of Houttuynia cordata on milk antioxidant indices

项目Items 对照组Control group 试验组Experimental group P值P-value
总抗氧化能力T-AOC/(mmol/mL) 0.99±0.03 0.97±0.01 0.486
1,1-二苯基-2-三硝基苯肼DPPH/(μg/mL) 5.06±0.23 5.10±0.13 0.879
谷胱甘肽GSH/(μmol/L) 69.52±8.25b 90.13±3.82a 0.047
总超氧化物歧化酶T-SOD/(U/mL) 106.96±12.46b 148.12±11.16a 0.028
丙二醛MDA/(nmol/mL) 3.36±0.60 3.08±0.39 0.695

2.3 鱼腥草对牛奶中代谢物组成的影响

2.3.1 多元变量模式识别分析

由图1-A可知,主成分分析(PCA)结果显示,牛奶样本均位于95%置信区间内,表明各组样本组内差异小,2个组的牛奶样本变化规律不同,说明鱼腥草使牛奶中的代谢物发生了不同幅度的变化,即奶牛摄入鱼腥草对牛奶代谢组有较大的影响。由图1-B可知,OPLS-DA得分图显示,2个组间牛奶样本均发生明显的分离,同一组内牛奶样本呈聚集趋势,模型参数R2Y=0.969,Q2=0.713,表明组间聚类差异显著,OPLS-DA模型稳定可靠,对2个组牛奶样品具有良好的预测能力。
图1 已鉴定代谢物的多元统计分析

A:主成分分析 PCA;B:正交偏最小二乘方-判别分析 OPLS-DA。

Fig.1 Multivariate statistical analysis of identified metabolites

2.3.2 牛奶差异代谢物的筛选

由图2可知,共筛选出了3 102个差异代谢物,其中磷酰胆碱、六氢吡啶、D-甘露糖-1-磷酸和甘露糖-6-磷酸等1 131个代谢物显著下调(P<0.05),N-乙酰神经氨酸、L-络氨酸和核黄素等1 931个代谢物显著上调(P<0.05)。
图2 差异代谢物火山图

Fig.2 Volcano map of differential metabolites

由表4可知,以VIP>1、P<0.05及差异倍数(FC)≥1.5或FC ≤0.67为筛选条件,共筛选出了23个共有差异代谢物,其中14个共有差异代谢物显著上调,分别为N-乙酰神经氨酸、N-乙酰半乳糖胺、D-葡萄糖醛酸、3-吡啶乙酸、L-酪氨酸、核黄素、L-氢化乳清酸、组氨酸、葡萄糖胺-1-磷脂酸、D-阿拉伯糖醇、D-木糖醇、葡萄糖酸和N-甲基-L-谷氨酸;9个共有差异代谢物显著下调,分别为磷酸烯醇丙酮酸、3-甲基胞苷、环磷酸腺苷、N-乙酰氨基葡萄糖-1-磷酸、甘露糖-1-磷酸、1-磷酸岩藻糖、甘露糖-6-磷酸、六氢吡啶和脱磷酸辅酶A。
表4 鱼腥草对牛奶中代谢物的影响

Table 4 Effects of Houttuynia cordata on metabolites in milk

代谢物
Metabolites		 差异倍数
FC		 P值
P-value		 变量投影重要性
VIP		 变化趋势
Change trend		 HMD KEGG
N-乙酰神经氨酸
N-acetylneuraminic acid		 1.56 0.024 1.54 上调 HMDB0000230 C00270
N-乙酰半乳糖胺
N-acetylgalactosamine		 3.35 0.001 2.45 上调 HMDB0000212 C01074
D-葡萄糖醛酸D-glucuronic acid 1.70 0.001 2.18 上调 HMDB0000127 C00191
3-吡啶乙酸3-pyridylacetic acid 3.89 0.001 2.32 上调 HMDB0001538
L-酪氨酸L-tyrosine 1.54 0.005 1.86 上调 HMDB0000158 C00082
核黄素Riboflavin 1.67 0.001 2.11 上调 HMDB0000244 C00255
L-氢化乳清酸
L-dihydroorotic acid		 1.52 0.019 1.55 上调 HMDB0003349 C00337
组氨酸Histidine 1.64 0.049 1.44 上调 HMDB0000177 C00135
葡萄糖胺-1-磷脂酸
Glucosamine-1-phosphate		 1.52 0.005 1.79 上调 HMDB0001109 C06156
D-阿拉伯糖醇D-arabitol 1.78 0.001 1.90 上调 HMDB0000568 C01904
D-木糖醇D-xylitol 1.78 0.001 1.90 上调 HMDB0002917 C00379
葡萄糖酸Gluconic acid 2.27 0.001 2.45 上调 HMDB0000625 C00257
N-甲基-L-谷氨酸
N-methy-L-glutamic acid		 1.63 0.001 2.09 上调 HMDB0062660 C01046
N-乙酰-D-葡萄糖胺
N-acetyl-D-glucosamine		 2.79 0.001 1.57 上调 HMDB0000215 C00140
磷酸烯醇丙酮酸
Phosphoenolpyruvic acid		 0.22 0.001 2.35 下调 HMDB0000263 C00074
5-甲基胞苷5-methylcytidine 0.59 0.001 1.92 下调 HMDB0000982
环磷酸腺苷
Adenosine 3',5'-cyclic phosphate		 0.47 0.001 1.99 下调 HMDB0000058 C00575
N-乙酰氨基葡萄糖-1-磷酸
N-acetyl-glucosamine-1-phosphate		 0.60 0.001 1.92 下调 HMDB0001367 C04256
D-甘露糖-1-磷酸
D-mannose-1-phosphate		 0.31 0.001 2.41 下调 HMDB0006330 C00636
1-磷酸岩藻糖
Fucose-1-phosphate		 0.42 0.047 1.23 下调 HMDB0001265 C02985
甘露糖-6-磷酸
Mannose-6-phosphate		 0.37 0.001 2.22 下调 HMDB0001078 C00275
六氢吡啶Piperidine 0.43 0.001 2.53 下调 HMDB0034301 C01746
脱磷辅酶A Dephospho-CoA 0.40 0.001 2.00 下调 HMDB0001373 C00882

HMD为该物质在HMDB数据库中的索引;KEGG为该物质在KEGG COMPOUND数据库中的索引。

HMD is the index of the substance in the HMDB database; KEGG is the index of the substance in the KEGG COMPOUND database.

2.3.3 KEGG代谢通路富集分析

使用MetaboAnalyst 6.0分析平台对对照组和试验组牛奶中筛选到的23个共有差异代谢物进行KEGG代谢通路富集分析,由图3可知,共有差异代谢物主要富集的代谢通路包括核黄素代谢、抗坏血酸和醛糖酸代谢、苯丙氨酸、酪氨酸和色氨酸的生物合成、组氨酸代谢、氨基酸糖和核苷酸糖代谢、果糖和甘露糖代谢、戊糖和葡萄糖醛酸互相转化、苯丙氨酸代谢、泛醌和其他萜类醌生物合成、柠檬酸循环(TCA循环)、泛酸和辅酶A生物合成、β-丙氨酸代谢、戊糖磷酸途径、丙酮酸代谢、糖酵解/糖异生、磷酸肌醇代谢、嘧啶代谢和酪氨酸代谢。
图3 差异代谢物KEGG富集通路气泡图

Fig.3 Bubble diagram of different metabolic KEGG enrichment pathways

由表5可知,与对照组相比,试验组在氨基酸糖和核苷酸糖代谢途径中N-乙酰半乳糖胺、N-乙酰-D-葡萄糖胺含量显著上升(P<0.05),1-磷酸岩藻糖、D-甘露糖-1-磷酸、甘露糖-6-磷酸和N-乙酰氨基葡萄糖-1-磷酸含量显著下降(P<0.05);在果糖和甘露糖代谢途径中甘露糖-6-磷酸、1-磷酸岩藻糖和D-甘露糖-1-磷酸含量显著下降(P<0.05);在戊糖和葡萄糖醛酸互相转化途径中D-阿拉伯糖醇和D-木糖醇含量显著上升(P<0.05);在苯丙氨酸、酪氨酸和色氨酸的生物合成代谢途径中L-酪氨酸含量显著上升(P<0.05);在核黄素代谢途径中核黄素含量显著上升(P<0.05)。
表5 代谢通路以及相关差异代谢物

Table 5 Metabolic pathways and associated differential metabolites

序号
No		 代谢途径
Metabolic pathways		 差异代谢物
Differential metabolites		 P值
P-value
1 氨基酸糖和核苷酸糖代谢 N-乙酰半乳糖胺↑、N-乙酰-D-葡萄糖胺↑、1-磷酸岩藻
糖↓、D-甘露糖-1-磷酸↓、甘露糖-6-磷酸↓、
N-乙酰氨基葡萄糖-1-磷酸↓		 <0.001
2 果糖和甘露糖代谢 甘露糖-6-磷酸↓、1-磷酸岩藻糖↓、
D-甘露糖-1-磷酸↓		 <0.001
3 戊糖和葡萄糖醛酸互相转化 D-阿拉伯糖醇↑、D-木糖醇↑ 0.014
4 苯丙氨酸、酪氨酸和色氨酸的生物合成 L-酪氨酸↑ 0.039
5 核黄素代谢 核黄素↑ 0.039
6 苯丙氨酸代谢 L-酪氨酸↑ 0.078
7 抗坏血酸和醛糖酸代谢 D-葡萄糖醛酸↑ 0.085
8 组氨酸代谢 组氨酸↑ 0.146
9 泛醌和其他萜类醌生物合成 L-酪氨酸↑ 0.162
10 柠檬酸循环(TCA循环) 磷酸烯醇丙酮酸↓ 0.179
11 泛酸和辅酶A生物合成 脱磷辅酶A↓ 0.179
12 β-丙氨酸代谢 脱磷辅酶A↓ 0.187
13 戊糖磷酸途径 葡萄糖酸↑ 0.203
14 丙酮酸代谢 磷酸烯醇丙酮酸↓ 0.203
15 糖酵解/糖异生 磷酸烯醇丙酮酸↓ 0.226
16 磷酸肌醇代谢 D-葡萄糖醛酸↑ 0.257
17 嘧啶代谢 L-氢化乳清酸↑ 0.321
18 酪氨酸代谢 L-酪氨酸↑ 0.341

↑:上升 increased;↓:下降 decreased。

2.4 鱼腥草对煮沸和巴氏杀菌牛奶储藏期间抗氧化指标的影响

由表6可知,与对照组相比,试验组煮沸牛奶储藏3和6 d的GSH含量以及储藏6 d的T-AOC显著增加(P<0.05),煮沸牛奶储藏3和6 d的MDA含量显著降低(P<0.05);试验组巴氏杀菌牛奶储藏3和6 d的T-AOC显著增加(P<0.05),巴氏杀菌牛奶储藏3和6 d的MDA含量显著降低(P<0.05)。
表6 鱼腥草对煮沸和巴氏杀菌牛奶储藏期间抗氧化指标的影响

Table 6 Effects of Houttuynia cordata on antioxidant indices during storage of boiled and pasteurized milk

项目
Items		 对照组
Control group		 试验组
Experimental group		 P值
P-value
煮沸(储藏3 d) Boil (storage for 3 days)
总抗氧化能力T-AOC/(mmol/mL) 1.18±0.04 1.08±0.06 0.168
1,1-二苯基-2-三硝基苯肼DPPH/(μg/mL) 1.84±0.06 1.89±0.06 0.579
谷胱甘肽GSH/(μmol/L) 51.33±2.15b 68.88±6.20a 0.026
丙二醛MDA/(nmol/mL) 8.32±0.66a 3.89±0.32b <0.001
煮沸(储藏6 d) Boil (storage for 6 days)
总抗氧化能力T-AOC/(mmol/mL) 0.75±0.02b 0.82±0.02a 0.034
1,1-二苯基-2-三硝基苯肼DPPH/(μg/mL) 0.57±0.02 0.58±0.01 0.641
谷胱甘肽GSH/(μmol/L) 38.25±1.48b 57.78±4.06a 0.002
丙二醛MDA/(nmol/mL) 11.74±1.12a 9.04±0.65b 0.045
巴氏杀菌(储藏3 d) Pasteurized (storage for 3 days)
总抗氧化能力T-AOC/(mmol/mL) 0.82±0.02b 0.96±0.01a <0.001
1,1-二苯基-2-三硝基苯肼DPPH/(μg/mL) 1.84±0.08 1.87±0.03 0.754
谷胱甘肽GSH/(μmol/L) 44.22±2.79 43.65±5.96 0.932
丙二醛MDA/(nmol/mL) 11.02±0.47a 4.56±0.53b <0.001
巴氏杀菌(储藏6 d) Pasteurized (storage for 6 days)
总抗氧化能力T-AOC/(mmol/mL) 0.81±0.02b 0.91±0.03a 0.015
1,1-二苯基-2-三硝基苯肼DPPH/(μg/mL) 0.52±0.01 0.48±0.02 0.192
谷胱甘肽GSH/(μmol/L) 42.69±7.59 41.04±4.09 0.852
丙二醛MDA/(nmol/mL) 14.00±0.54a 8.45±0.15b <0.001

3 讨论

3.1 鱼腥草对牛奶抗氧化能力的影响

牛奶和奶制品含有多种对生理和生化功能有益的生物活性物质,例如蛋白质、钙、必需脂肪酸、氨基酸和脂肪,是人类抗氧化化合物的主要来源。牛奶氧化会引起强烈的异味和牛奶营养质量的恶化,牛奶和乳制品的氧化稳定性是牛奶中抗氧化和促氧化过程之间微妙平衡的结果。牛奶中抗氧化成分的含量和抗氧化能力可以通过调整饲粮组成进行调控。研究表明,在奶牛的饲粮中添加南瓜青贮饲料可显著提高初乳的营养价值,包括类胡萝卜素的含量,初乳的抗氧化能力也明显更高;对饲喂富含类胡萝卜素奶牛的牛奶研究也得出了类似的结果[13]。Santos等[14]报道,采食含有葡萄渣青贮饲料奶牛的牛奶具有更高的抗氧化能力。鱼腥草中的绿原酸、多糖、多酚和黄酮类活性物质具有较强的抗氧化能力[15]。本研究发现,饲喂鱼腥草显著增加了牛奶中T-SOD活性和GSH含量。超氧化物歧化酶(SOD)可以催化超氧自由基转化为过氧化氢和氧气,起到清除超氧自由基的作用,而GSH是一种还原性强的抗氧化剂,能直接清除自由基,还作为辅酶在谷胱甘肽过氧化物酶(GSH-Px)中发挥作用,将过氧化氢转化为水,从而缓解氧化损伤[16]。祝璟琳等[17]研究表明,鱼腥草能缓解无乳链球菌引起的尼罗罗非鱼肝脏损伤以及改善氧化还原状态。同样,给大鲟幼鱼饲喂0.5 g/kg剂量的含鱼腥草的复方中草药显著增加了其血清中SOD活性和GSH含量[18]。在大鼠饲粮中添加400 mg/kg的鱼腥草显著增加了大鼠肝脏、胰腺和脂肪组织中T-SOD活性;此外,添加相同剂量的鱼腥草还可降低链脲佐菌素诱导的大鼠线粒体氧化应激和糖尿病诱导的血液中MDA含量[19]。Wang等[20]研究发现,在饲粮中添加1 000 mg/kg的鱼腥草水提取物显著提高了兔背最长肌中抗氧化相关基因的mRNA表达水平。何晓芳等[21]研究发现,在海兰褐壳蛋鸡饲粮中添加鱼腥草显著增加了十二指肠和空肠黏膜中T-AOC和SOD活性。司雨豪[22]研究发现,鱼腥草粉(2%)可以显著增加仔猪血清T-AOC、SOD活性和GSH含量,显著降低MDA含量[22]。Wang等[23]研究发现,鱼腥草的水蒸气提取物可以通过增加奶牛乳腺组织中血红素加氧酶-1含量,增强抗氧化能力,从而降低患乳腺炎的概率。本研究结果与前人研究结果一致,表明饲喂鱼腥草可作为提升牛奶抗氧化能力的有效营养干预策略。

3.2 鱼腥草对牛奶货架期的影响

牛奶和乳制品的货架期在一定程度上取决于其中抗氧化物质含量的多少以及抗氧化能力的强弱。非酶性抗氧化物质如GSH含量的提高以及SOD、过氧化氢酶(CAT)、GSH-Px等活性增强可延长牛奶及乳制品的货架期[24-25]。饲喂多酚类、黄酮类和维生素等活性物质可提升牛奶抗氧化能力,延长货架期。本试验发现,试验组牛奶中总酚和总黄酮含量显著高于对照组,表明奶牛摄入鱼腥草可能通过提高多酚和黄酮在牛奶中的沉积来提升牛奶的抗氧化能力。奶牛饲粮中添加富含酚类或黄酮类化合物的青贮果渣可促进牛奶中酚类或黄酮的蓄积,提高牛奶的抗氧化能力[26-27]。多酚和黄酮因其抗氧化活性,常添加于肉制品以延长其货架期[28]。研究表明,牛肉中添加含多酚的柑橘渣及含黄酮醇的葡萄渣可以延长其货架期[29]。此外,肉牛饲粮中添加富含黄酮和多酚的葡萄渣可提高储藏3、7和10 d牛肉中GSH含量,并且降低储藏3和7 d牛肉中MDA含量,进而延长了牛肉的货架期[30]。本试验中,奶牛采食鱼腥草显著增强了煮沸牛奶储藏3和6 d的GSH含量以及储藏6d的T-AOC,显著降低了煮沸牛奶储藏3和6 d的MDA含量;奶牛采食鱼腥草显著增强了巴氏杀菌牛奶储藏3和6 d的T-AOC活性,显著降低了巴氏杀菌牛奶储藏3和6 d中的MDA含量。T-AOC可反映食品中酶类和非酶类主导的抗氧化系统作用下的总体抗氧化能力,而MDA是脂质非酶性氧化作用下的终产物,其含量增加会加速食品的脂质氧化反应[31]。本试验中,试验组牛奶中MDA含量显著降低,可能与牛奶的抗氧化能力增加有关。煮沸和巴氏杀菌牛奶中主要抗氧化物质存在差异,可能与温度的升高会改变牛奶中抗氧化物质的含量有关[32]。综上所述,鱼腥草中的活性成分多酚和黄酮可以通过提高牛奶的抗氧化能力来延长牛奶的货架期。

3.3 牛奶中代谢物对抗氧化能力和货架期的影响

牛奶的抗氧化能力与其含有的抗氧化成分含量直接相关。在正常新陈代谢的情况下,生物体中产生的自由基会被抗氧化系统中和。代谢紊乱导致自由基和抗氧化反应之间的平衡紊乱,从而导致细胞中的自由基过量积累,天然抗氧化物质有助于延长食品货架期。在自然界中多酚通常以糖苷的形式存在,这有利于降低其脂溶性。糖苷的水解通常由乳糖酶根皮苷水解酶催化,释放出亲脂性更强的糖苷配基[33],然后糖苷配基能够通过被动扩散进入肠上皮细胞。或者,糖苷可以通过钠依赖性葡萄糖转运蛋白主动转运到肠上皮细胞中,然后被胞质β-葡萄糖苷酶水解[34],使其通过肠道进入循环系统被机体吸收。因此,虽然糖不具备抗氧化作用,但与多酚结合后却能增强多酚的抗氧化效果,这种协同作用可以提高食品的抗氧化能力,延缓食品的氧化过程。在食品加工中,糖和多酚常常被用于一起,如糖和水果中的花青素结合在一起形成的复合物等。这种组合不仅可以提高食品的口感和口味,还能增加食品的营养价值和抗氧化能力[35]。D-阿拉伯糖醇和D-木糖醇等多元醇是一类具有多个羟基(-OH)基团的有机化合物,可以通过抑制氧化反应和减少自由基对食品的氧化损伤,保持食品的新鲜度和质量,延长其保存时间[36]。此外,D-木糖醇因具有抑制微生物繁殖和防止蛋白质变性的特性,在食品工业中被广泛用作防腐剂和抗氧化剂[37]。本研究发现,奶牛摄入鱼腥草后在氨基酸糖和核苷酸糖代谢途径中N-乙酰半乳糖胺、N-乙酰-D-葡萄糖胺含量显著上升,在戊糖和葡萄糖醛酸互相转化途径中D-阿拉伯糖醇和D-木糖醇含量显著上升,表明鱼腥草中的多酚可能通过调节糖代谢提高牛奶的货架期。
牛奶中的蛋白质也具有抗氧化特性,因为其含有的含硫氨基酸以及色氨酸、酪氨酸和苯丙氨酸可以通过中和自由基充当抗氧化剂。牛奶和乳制品中含有的特异性蛋白质片段生物活性肽具有清除自由基、螯合金属离子和抑制脂质过氧化的能力[38]。研究表明,肽的氨基酸组成与抗氧化活性相互作用,肽通常由5~11个氨基酸残基组成,包括疏水性残基(脯氨酸、组氨酸、酪氨酸、色氨酸或半胱氨酸),其游离形式也具有抗氧化活性[39]。本研究发现,奶牛在苯丙氨酸、酪氨酸和色氨酸的生物合成代谢途径中L-酪氨酸含量显著上升,表明鱼腥草可能通过增强氨基酸代谢提高牛奶的货架期。牛奶中的脂溶性维生素具有清除自由基、抑制乳脂氧化的作用,此外,B族维生素及维生素C(抗坏血酸钠)也可以中和牛奶中的自由基,延缓牛奶的氧化过程,减少氧化反应对牛奶的影响,保持牛奶的新鲜度和营养价值[40-41]。本研究发现,在核黄素代谢途径中核黄素含量显著上升。以上结果表明,饲喂奶牛鱼腥草有助于增加提高牛奶抗氧化能力和延长货架期的代谢物含量。

4 结论

奶牛饲粮中添加2%鱼腥草可提高牛奶中总黄酮和总酚含量,增强鲜奶和热处理后不同冷藏时间牛奶的抗氧化能力,并可在一定程度上延长牛奶的货架期,这与牛奶中抗氧化代谢物含量增加有关。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
MILLOGO V, SISSAO M, SIDIBÉ A G, et al. Effect of storage time and temperature on raw milk composition of dairy cattle in tropical conditions[J]. African Journal of Dairy Farming and Milk Production, 2015, 2(1):104-108.

[2]
PATIL S S, ZAGARE M S, GHORADE I B, et al. Isolation of pathogens from dairy products and effect of temperature on pathogens[J]. Paripex-Indian Journal of Research, 2014, 3(1):76-79.

[3]
殷献华, 殷诚, 钱静, 等. 鲜牛奶品质参数的动力学特性研究及货架期预测[J]. 包装工程, 2024, 45(3):55-61.

YIN X H, YIN C, QIAN J, et al. Kinetics and shelf life prediction of fresh milk quality parameters[J]. Packaging Engineering, 2024, 45(3):55-61. (in Chinese)

[4]
RITOTA M, DI COSTANZO M G, MATTERA M, et al. New trends for the evaluation of heat treatments of milk[J]. Journal of Analytical Methods in Chemistry, 2017, 2017(1):1864832.

[5]
BEZIE A. The effect of different heat treatment on the nutritional value of milk and milk products and shelf-life of milk products.A review[J]. Journal of Dairy&Veterinary Sciences, 2019, 11(5):555822.

[6]
KUMNERDKHONKAEN P, SAENGLEE S, ASGAR M A, et al. Antiproliferative activities and phenolic acid content of water and ethanolic extracts of the powdered formula of Houttuynia cordata Thunb.fermented broth and Phyllanthus emblica Linn. fruit[J]. BMC Complementary and Alternative Medicine, 2018, 18(1):130.

[7]
MARCHAND S, HEYLEN K, MESSENS W, et al. Seasonal influence on heat-resistant proteolytic capacity of Pseudomonas lundensis and Pseudomonas fragi,predominant milk spoilers isolated from Belgian raw milk samples[J]. Environmental Microbiology, 2009, 11(2):467-482.

[8]
YUAN L, SADIQ F A, BURMØLLE M, et al. Insights into psychrotrophic bacteria in raw milk:a review[J]. Journal of Food Protection, 2019, 82(7):1148-1159.

[9]
SAIKIA R R, BORPUZARI T, HAZARIKA M, et al. Evaluation of the effect of plant extracts (Paederia foetida and Houttuynia cordata) incorporation on the quality of masala paneer[J]. International Journal of Current Microbiology and Applied Sciences, 2020, 9(12):240-249.

[10]
WEI P P, LUO Q, HOU Y, et al. Houttuynia cordata Thunb.:a comprehensive review of traditional applications,phytochemistry,pharmacology and safety[J]. Phytomedicine, 2024,123:155195.

[11]
KANG C, LEE H, HAH D Y, et al. Protective effects of houttuynia cordata thunb. on gentamicin-induced oxidative stress and nephrotoxicity in rats[J]. Toxicological research, 2013, 29(1):61-67.

DOI PMID

[12]
MA F T, SHAN Q, JIN Y H, et al. Effect of Lonicera japonica extract on lactation performance,antioxidant status,and endocrine and immune function in heat-stressed mid-lactation dairy cows[J]. Journal of Dairy Science, 2020, 103(11):10074-10082.

[13]
ANTONE U, ZAGORSKA J, STERNA V, et al. Effects of dairy cow diet supplementation with carrots on milk composition,concentration of cow blood serum carotenes,and butter oil fat-soluble antioxidative substances[J]. Agronomy Research, 2015, 13(4):879-891.

[14]
SANTOS N W, SANTOS G T D, SILVA-KAZAMA D C, et al. Production,composition and antioxidants in milk of dairy cows fed diets containing soybean oil and grape residue silage[J]. Livestock Science, 2014,159:37-45.

[15]
HAMED M, ABDALLAH I A, BEDAIR A, et al. Sample preparation methods for determination of quercetin and quercetin glycosides in diverse matrices[J]. Microchemical Journal, 2023,194:109233.

[16]
WANG W J, ZHANG Z W, LIU X Y, et al. An improved GC-MS method for malondialdehyde (MDA) detection:avoiding the effects of nitrite in foods[J]. Foods, 2022, 11(9):1176.

[17]
祝璟琳, 季桓涛, 肖炜, 等. 鱼腥草对无乳链球菌引起吉富罗非鱼肝脏损伤的修复作用[J]. 水产学报, 2020, 44(7):1187-1197.

ZHU J L, JI H T, XIAO W, et al. Protective effects from Houttuynia cordata against streptococcus agalactiae induced liver injury of GIFT Oreochromis niloticus strain[J]. Journal of Fisheries of China, 2020, 44(7):1187-1197

[18]
张建明, 张德志, 舒德斌, 等. 4种中药复方对长江鲟幼鱼抗氧化和抗应激指标的影响[J]. 南方农业学报, 2022, 53(2):568-576.

ZHANG J, ZHANG D, SHU D, et al. Effects of four Chinese herbal medicine compound on antioxidation and anti-stress indexes of Acipenser dabryanus juveniles[J]. Journal of Southern Agriculture, 2022, 53(2):568-576.

[19]
KUMAR M, PRASAD S K, KRISHNAMURTHY S, et al. Antihyperglycemic activity of Houttuynia cordata Thunb. in streptozotocin-induced diabetic rats[J]. Advances in Pharmacological Sciences, 2014,2014:809438.

[20]
WANG X, XIE S L, CHEN R, et al. Dietary Houttuynia cordata Thunb. extract supplementation influences on antioxidant enzymes,meat quality,and gene expressions in fresh rabbit Longissimus thoracis et lumborum meat[J]. Meat Science, 2025,226:109845.

[21]
何晓芳, 韦春秀, 张新月, 等. 菊粉和鱼腥草素联用对海兰褐蛋雏鸡生长性能、肠道消化酶活性及抗氧化性能的影响[J]. 饲料研究, 2022(19):53-57.

HE X F, WEI C X, ZHANG X Y, et al. Effect of inulin and Houttuynia cordata on growth performance,intestinal digestive enzyme activity,and anti-oxidation of Hyline brown chick[J]. Feed Research, 2022(19):53-57.

[22]
司雨豪. 鱼腥草对断奶清平仔猪生长性能、血液理化指标与肠道健康的影响[D].硕士学位论文. 武汉: 华中农业大学, 2024.

SI Y H. The effects of houttuynia cordata on the growth performance,blood physicochemical indicators,and intestinal health of weaned Qingping piglets[D].Master’s Thesis. Wuhan: Huazhong Agricultural University, 2024. (in Chinese)

[23]
WANG N, ZHU Y B, LI D D, et al. 2-methyl nonyl ketone from Houttuynia cordata Thunb. alleviates LPS-induced inflammatory response and oxidative stress in bovine mammary epithelial cells[J]. Frontiers in Chemistry, 2022,9:793475.

[24]
USTA B, YILMAZ E L Antioxidant enzymes of milk and their biological effects[J]. Ziraat Fakültesi Dergisi,Uludağ Üniversitesi, 2013, 27(2):123-130.

[25]
KHAN I T, NADEEM M, IMRAN M, et al. Antioxidant properties of milk and dairy products:a comprehensive review of the current knowledge[J]. Lipids in Health and Disease, 2019, 18(1):41.

[26]
NIERO G, PENASA M, CURRÒ S, et al. Development and validation of a near infrared spectrophotometric method to determine total antioxidant activity of milk[J]. Food Chemistry, 2017,220:371-376.

[27]
LEE S H, HUMPHRIES D J, COCKMAN D A, et al. Accumulation of citrus flavanones in bovine milk following citrus pulp incorporation into the diet of dairy cows[J]. EC Nutrition, 2017, 7(4):143-154.

[28]
MUNEKATA P E S, PATEIRO M, BELLUCCI E R B, et al. Chapter five-strategies to increase the shelf life of meat and meat products with phenolic compounds[J]. Advances in Food and Nutrition Research, 2021,98:171-205.

[29]
TAYENGWA T, CHIKWANHA O C, GOUWS P, et al. Dietary citrus pulp and grape pomace as potential natural preservatives for extending beef shelf life[J]. Meat Science, 2020,162:108029.

[30]
MOLOSSE V L, DEOLINDO G L, LAGO R V, et al. The use of secondary grape biomass in beef cattle nutrition on carcass characteristics,quality and shelf life of meat[J]. Food and Nutrition Sciences, 2024, 15(6):447-469.

[31]
ZENG Z Y, HE X, LI C Y, et al. Oral delivery of antioxidant enzymes for effective treatment of inflammatory disease[J]. Biomaterials, 2021,271:120753.

[32]
CHÁVEZ-SERVÍN J L, ANDRADE-MONTEMAYOR H M, VELÁZQUEZ VÁZQUEZ C, et al. Effects of feeding system,heat treatment and season on phenolic compounds and antioxidant capacity in goat milk,whey and cheese[J]. Small Ruminant Research, 2018,160:54-58.

[33]
DAY A J, CAÑADA F J, DÍAZ J C, et al. Dietary flavonoid and isoflavone glycosides are hydrolysed by the lactase site of lactase phlorizin hydrolase[J]. FEBS Letters, 2000, 468(2/3):166-170.

[34]
GEE J M, DUPONT M S, DAY A J, et al. Intestinal transport of quercetin glycosides in rats involves both deglycosylation and interaction with the hexose transport pathway[J]. The Journal of Nutrition, 2000, 130(11):2765-2771.

[35]
HANUKA KATZ I, ERAN NAGAR E, OKUN Z, et al. The link between polyphenol structure,antioxidant capacity and shelf-life stability in the presence of fructose and ascorbic acid[J]. Molecules, 2020, 25(1):225.

[36]
RICE T, ZANNINI E, K ARENDT E, et al. A review of polyols-biotechnological production,food applications,regulation,labeling and health effects[J]. Critical Reviews in Food Science and Nutrition, 2020, 60(12):2034-2051.

[37]
UMAI D, KAYALVIZHI R, KUMAR V, et al. Xylitol:bioproduction and applications-a review[J]. Frontiers in Sustainability, 2022,3:826190.

[38]
STOBIECKA M, KRÓL J, BRODZIAK A. Antioxidant activity of milk and dairy products[J]. Animals, 2022, 12(3):245.

[39]
UDENIGWE C C, ALUKO R E. Food protein-derived bioactive peptides:production,processing,and potential health benefits[J]. Journal of Food Science, 2012, 77(1):R11-R24.

[40]
RASHAD M M, MAHMOUD A E, ABDOU H M, et al. Improvement of nutritional quality and antioxidant activities of yeast fermented soybean curd residue[J]. African Journal of Biotechnology, 2011, 10(28):5504-5513.

[41]
MANN S, SHANDILYA U K, SODHI M, et al. Determination of antioxidant capacity and free radical scavenging activity of milk from native cows (Bos indicus),exotic cows (Bos taurus),and riverine buffaloes (Bubalus bubalis) across different lactation stages[J]. International Journal of Dairy Science & Processing, 2016, 3(4):66-70.

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