1 材料与方法
1.1 主要仪器和试剂
1.2 试验动物和试验设计
1.3 UPLC-MS/MS检测条件
1.4 数据处理与统计分析
1.5 差异代谢物筛选与生物学功能分析
2 结果
2.1 代谢物鉴定及多元统计分析
图1 已鉴定代谢物的多元统计分析图a为PCA,图b为C0与T0组OPLS-DA,图c为C21与T21组OPLS-DA。C0组和T0组表示分娩当天的对照组和试验组,C21组和T21组表示产后第21天的对照组和试验组。 Fig.1 Multivariate statistical analysis of identified metabolites Figure amean PCA, figure b mean OPLS-DA of C0 and T0 groups, and figure c mean OPLS-DA of C21 and T21 groups. Group C0 and group T0 represented the control group and the experimental group on day of calving, and group C21 and group T21 represented the control group and the experimental group on day 21 after calving. |
2.2 差异代谢物筛选及聚类分析
表1 分娩当天C0组(对照组)与T0组(试验组)筛选的差异代谢物Table 1 Differential metabolites between C0 group (control group) and T0 group (experimental group) on day of calving |
| 代谢物 Metabolites | 变量投影重要度 VIP | P值 P-value | 差异倍数 FC | 变化趋势 Variation trend |
|---|---|---|---|---|
| 氨基酸及其衍生物(10种) Amino acids and their derivatives (10 kinds) | ||||
| L-苯丙氨酸 L-Phe | 2.06 | 0.009 | 11.18 | 上调 |
| 谷氨酸-酪氨酸 Glu-Tyr | 1.65 | 0.005 | 1.53 | 上调 |
| 精氨酸-精氨酸 Arg-Arg | 1.36 | 0.027 | 0.81 | 下调 |
| 谷氨酸-异亮氨酸 Glu-Ile | 1.56 | 0.012 | 0.75 | 下调 |
| L-脯氨酸 L-Pro | 1.16 | 0.044 | 0.69 | 下调 |
| 亮氨酸-苯丙氨酸 Leu-Phe | 2.00 | 0.001 | 0.67 | 下调 |
| 亮氨酸-亮氨酸-正亮氨酸 Leu-Leu-norleucine | 1.74 | 0.009 | 0.59 | 下调 |
| 谷氨酸-S-甲基半胱氨酰-丙氨酸 Glu-S-methylcysteinyl-Ala | 2.16 | <0.001 | 0.46 | 下调 |
| 代谢物 Metabolites | 变量投影重要度 VIP | P值 P-value | 差异倍数 FC | 变化趋势 Variation trend |
| 甲硫氨酸-酪氨酸 Met-Tyr | 2.51 | <0.001 | 0.38 | 下调 |
| L-异亮氨酸 L-Ile | 1.55 | 0.027 | 0.08 | 下调 |
| 碳水化合物(3种) Carbohydrates (3 kinds) | ||||
| 磷酸羟基丙酮酸 Phosphohydroxypyruvic acid | 1.64 | 0.013 | 3.26 | 上调 |
| 半乳糖-1-磷酸 Galactose-1-phosphate | 1.63 | 0.001 | 2.45 | 上调 |
| α-乳糖 α-lactose | 1.57 | 0.011 | 1.64 | 上调 |
| 脂质(4种) Lipids (4 kinds) | ||||
| 咖啡酸 Caffeic acid | 1.13 | 0.049 | 1.40 | 上调 |
| 血栓素B2 TXB2 | 1.60 | 0.010 | 0.67 | 下调 |
| 己二酸 Adipic acid | 1.37 | 0.022 | 0.62 | 下调 |
| 植物鞘氨醇 Phytosphingosine | 2.07 | 0.001 | 0.51 | 下调 |
| 其他代谢物(3种) Other metabolites (3 kinds) | ||||
| 鹅去氧胆酸 CDCA | 1.53 | 0.030 | 1.44 | 上调 |
| 黄嘌呤核苷酸 XMP | 2.15 | <0.001 | 0.62 | 下调 |
| 环磷酸鸟苷 cGMP | 2.34 | <0.001 | 0.45 | 下调 |
表2 产后第21天C21组(对照组)与T21组(试验组)筛选的差异代谢物Table 2 Differential metabolites between C21 group (control group) and T21 group (experimental group) on day 21 after calving |
| 代谢物 Metabolites | 变量投影重要度 VIP | P值 P-value | 差异倍数 FC | 变化趋势 Variation trend |
|---|---|---|---|---|
| 氨基酸及其衍生物(10种) Amino acids and their derivatives (10 kinds) | ||||
| L-酪氨酸 L-Tyr | 1.16 | 0.041 | 2.21 | 上调 |
| 天冬氨酸-丙氨酸 Asp-Ala | 1.47 | 0.014 | 0.84 | 下调 |
| 精氨酸-精氨酸 Arg-Arg | 2.07 | <0.001 | 0.66 | 下调 |
| 亮氨酸-苯丙氨酸 Leu-Phe | 2.14 | <0.001 | 0.65 | 下调 |
| 谷氨酸-异亮氨酸 Glu-Ile | 2.03 | <0.001 | 0.65 | 下调 |
| 谷氨酰胺-色氨酸 Gln-Try | 1.97 | <0.001 | 0.63 | 下调 |
| 甲硫氨酸-酪氨酸 Met-Tyr | 1.76 | 0.007 | 0.62 | 下调 |
| 异亮氨酸-酪氨酸 Ile-Tyr | 1.76 | 0.001 | 0.59 | 下调 |
| 亮氨酸-亮氨酸-正亮氨酸 Leu-Leu-norleucine | 2.23 | <0.001 | 0.55 | 下调 |
| 苯丙氨酸-苏氨酸 Phe-Thr | 1.08 | 0.021 | 0.55 | 下调 |
| 碳水化合物(1种) Carbohydrates (1 kind) | ||||
| 葡萄糖-6-磷酸 Glucose-6-phosphate | 1.38 | 0.033 | 1.16 | 上调 |
| 脂质(3种) Lipids (3 kinds) | ||||
| 植物鞘氨醇 Phytosphingosine | 2.20 | <0.001 | 0.64 | 下调 |
| 咖啡酸 Caffeic acid | 1.36 | 0.019 | 0.60 | 下调 |
| 白三烯A4 Leukotriene A4 | 2.47 | <0.001 | 0.24 | 下调 |
| 其他代谢物(6种) Other metabolites (6 kinds) | ||||
| 烟酰胺 Niacinamide | 1.26 | 0.009 | 2.11 | 上调 |
| 二磷酸尿核苷 UDP | 1.35 | 0.019 | 1.09 | 上调 |
| 鹅去氧胆酸 CDCA | 1.11 | 0.026 | 0.81 | 下调 |
| 脱氧尿苷 Deoxyuridine | 1.66 | 0.001 | 0.76 | 下调 |
| 黄嘌呤核苷酸 XMP | 2.29 | <0.001 | 0.51 | 下调 |
| 环磷酸鸟苷 cGMP | 2.39 | <0.001 | 0.24 | 下调 |
图2 差异代谢物聚类热图图a为C0与T0组的差异代谢物聚类热图,图b为C21与T21组的差异代谢物聚类热图。C0组和T0组表示分娩当天的对照组和试验组,C21组和T21组表示产后第21天的对照组和试验组。 Fig.2 Cluster heatmap of differential metabolites Figure a was the cluster heatmap of differential metabolites between C0 group and T0 group, and figure b was the cluster heatmap of differential metabolites between C21 group and T21 group. Group C0 and group T0 represented the control group and the experimental group on day of calving, and group C21 and group T21 represented the control group and the experimental group on day 21 after calving. |
2.3 差异代谢物的代谢通路富集分析
表3 分娩当天和产后第21天差异代谢物富集通路Table 3 Enrichment pathway of differential metabolites on day of calving and day 21 after calving |
| 通路 Pathway | P值 P-value | Impact值 Impact value |
|---|---|---|
| 分娩当天 On day of calving | ||
| 苯丙氨酸代谢 Phenylalanine metabolism | 0.11 | 0.36 |
| 半乳糖代谢 Galactose metabolism | 0.02 | 0.14 |
| 产后第21天 On day 21 after calving | ||
| 苯丙氨酸、酪氨酸和色氨酸生物合成 Phenylalanine, tyrosine and tryptophan biosynthesis | 0.03 | 0.50 |
| 烟酸和烟酰胺代谢 Nicotinate and nicotinamide metabolism | 0.10 | 0.19 |
| 酪氨酸代谢 Tyrosine metabolism | 0.29 | 0.14 |
| 淀粉和蔗糖代谢 Starch and sucrose metabolism | 0.13 | 0.14 |
| 嘧啶代谢 Pyrimidine metabolism | 0.03 | 0.10 |
Impact值为通路拓扑分析的影响值。 | |
Impact value was impact value of pathway topology analysis. |

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