基于LC-MS技术的海、淡水养殖刀鲚卵巢的代谢组学比较分析

高淑芳; 张金鹏; 施永海; 袁新程; 刘其根

doi:10.12131/20210185

基于LC-MS技术的海、淡水养殖刀鲚卵巢的代谢组学比较分析

doi: 10.12131/20210185

1.
上海海洋大学/水产科学国家级实验教学示范中心/农业农村部鱼类营养与环境生态研究中心/农业农村部淡水水产种质资源重点实验室, 上海 201306
2.
上海市水产研究所/上海市水产技术推广站, 上海 200433

基金项目: 农业部鱼类营养与环境生态研究中心 (A1-3201-19-300204)；上海市科委重点攻关项目(17391900300)

详细信息

作者简介:
高淑芳 (1994—)，女，硕士研究生，研究方向为刀鲚分子生物学。E-mail: sfgao101@163.com

通讯作者:
刘其根 (1965—)，男，教授，从事水域生态学研究。E-mail: qgliu@shou.edu.cn

中图分类号: S 965.227
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- 文章访问数: 34
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- 被引次数: 0
出版历程
- 收稿日期: 2021-06-24
- 修回日期: 2021-10-09
- 录用日期: 2021-11-04
- 网络出版日期: 2021-11-11

Metabonomics analysis of ovaries of Coilia nasus in seawater and freshwater based on liquid chromatography-mass spectrometry

1.
Shanghai Ocean University/National Demonstration Center for Experimental Fisheries Science Education/Centre for Research on Environmental Ecology and Fish Nutrition, Ministry of Agriculture and Rural Affairs/Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai 201306, China
2.
Shanghai Fisheries Research Institute/Shanghai Fisheries Technology Extension Station, Shanghai 200433, China

摘要

摘要: 为探究海、淡水养殖环境对刀鲚 (Coilia nasus) 卵巢发育的影响，该研究采用非靶向代谢组学的方法，测定了海、淡水环境下刀鲚卵巢中代谢产物的差异情况，并与基因组百科全书 (KEGG) 数据库进行比对，找出相对应的代谢通路并分析其原因。结果表明，海水组和淡水组样品共鉴定出47种差异代谢物 (P<0.05、FC>1、VIP>1)，与海水组相比，淡水组表达差异倍数最明显的为碳环血氧烷A2 (Carbocyclic Thromboxane A2)、半乳糖神经酰胺(Galactosyl ceramide)，差异倍数分别为10.40、2.78倍；与海水组相比，淡水组卵巢组织内皮质醇升高了1.61倍；对47种差异代谢物进行KEGG分析发现，变化显著的通路有氨酰-tRNA的生物合成和嘧啶代谢通路 (P<0.05)，皮质醇、氨酰-tRNA的生物合成通路、嘧啶代谢通路和鞘磷脂代谢通路可能与刀鲚生殖洄游过程中卵巢发育有关。
- 刀鲚 /
- 液相色谱-质谱联用 /
- 盐度 /
- 卵巢 /
- 代谢组学 /
- 代谢通路
Abstract: In order to clarify the effects of seawater and freshwater on the ovary development of Coilia nasus, we analyzed their differences by using non-targeted metabolomics, and compared with database of KEGG directly to find out the corresponding metabolic pathways, then analyzed its causes. The results show that a total of 47 metabolites had significant difference between the two groups (P<0.05, FC>1, VIP>1). Compared with the seawater group, the most significant differences in the expression were Carbocyclic Thromboxane A2, Galactosylceramide, and their differences were 10.40, 2.78 times, respectively. The cortisol in the ovarian tissue of the freshwater group increased by 1.61 times. According to the analysis on KEGG metabolic pathways of 47 different metabolites, the biosynthesis of aminoacyl-tRNA and pyridine metabolic pathways changed in the seawater and freshwater environments significantly (P<0.05). The biosynthesis pathway of cortisol, aminoyl-tRNA, pyrimidine metabolism pathway and sphingo-lipid metabolism pathway may be related to the ovarian development during the reproductive migration of C. nasus.
- Coilia nasus /
- LC-MS /
- Salinity /
- Ovary /
- Metabolomics /
- Metabolic pathways

HTML全文

图 1 海水组和淡水组的主成分分析 (a) 和正交偏最小二乘判别分析 (b) 得分图

Figure 1. PCA (a) and OPLS-DA (b) scores of seawater and freshwater

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图 2 置换检验图

Figure 2. Permutation test

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图 3 代谢产物Venn图

ａ.阳离子模式；b. 阴离子模式。

Figure 3. Venn map of metabolites

a. Positive ion mode; b. Negative ion mode.

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图 4 差异代谢物筛选火山图

Figure 4. Volcanic map of different metabolites

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图 5 代谢物的通路富集分析

a. 氨酰-tRNA生物合成；b. 嘧啶代谢；c. 鞘磷脂代谢；d. 三羧酸循环。

Figure 5. Metabolite pathway enrichment analysis

a. Aminoacyl-tRNA biosynthesis; b. Pyrimudine metabolism; c. Sphingolipid metabolism; d. Citrate cycle TCA.

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表 1 差异代谢物信息表

Table 1. Differential metabolites information sheet

差异代谢物　　　 Differential metabolite	分子式 Molecular formula	变量投影重要度 VIP	FC Fold change	P	变化趋势 Variation trend
磷脂酰肌醇 PI (20:4(5Z,8Z,11Z,14Z)/0:0)	C₂₉H₄₉O₁₂P	1.58	0.93	0.017	下降
谷氨酰胺色氨酸 Glutaminyltryptophan	C₁₆H₂₀N₄O₄	1.05	0.96	0.018	下降
L-异亮氨酸 L-Isoleucine	C₆H₁₃NO₂	1.33	0.96	0.005	下降
木麻黄6-α-D-葡萄糖苷 Casuarine 6-alpha-D-glucoside	C₁₄H₂₅NO₁₀	1.13	0.94	0.043	下降
甘油一脂 MG(10:0/0:0/0:0)	C₁₃H₂₆O₄	3.59	0.71	0.025	下降
皮质醇 Cortisol	C₂₁H₃0O₅	3.28	1.61	0.008	上升
碳环血氧烷A2 Carbocyclic thromboxane A2	C₂₂H₃6O₃	2.88	10.40	0.007	上升
(±)9-十八碳二烯酸 (±)9-HPODE	C₁₈H₃₂O₄	2.90	0.58	0.020	下降
N-棕榈酰蛋氨酸 N-palmitoyl methionine	C₂₁H₄₁NO₃S	1.86	0.75	0.022	下降
13-羟基十八酸 13-hydroxyoctadecanoic acid	C₁₈H₃₆O₃	1.95	0.86	0.005	下降
咖啡酰环戊醇 Caffeoylcycloartenol	C₃₉H₅₆O₄	2.07	0.81	0.036	下降
11,13-二十碳二烯酸 15-OxoEDE	C₂₀H₃₄O₃	2.51	0.70	0.032	下降
2,3-二氢苯并呋喃 2,3-dihydrobenzofuran	C₈H₈O	1.19	1.08	0.033	上升
4-甲酰基吲哚 4-formyl indole	C₉H₇NO	1.34	0.83	0.040	下降
红花素C Safflomin C	C₃₀H₃₀O₁₄	2.04	0.85	0.003	下降
6-[(2-羧基乙酰基)氧]-3,4,5-三羟基氧烷-2-羧酸 6-[(2-carboxyacetyl)oxy]-3,4,5-trihydroxyoxane-2-carboxylic acid	C₉H₁₂O₁₀	2.07	1.12	0.001	上升
异戊二烯 Isoputreanine	C₇H₁₆N₂O₂	2.33	0.84	0.001	下降
脯氨酸 L-Proline	C₅H₉NO₂	1.80	0.91	0.003	下降
羟脯氨酸 Hydroxyprolyl-hydroxyproline	C₁₀H₁₆N₂O₅	3.13	0.72	0.007	下降
5-O-阿魏酰黑精 5-O-Feruloylnigrumin	C₂₁H₂₅NO₁₀	1.0	0.95	0.037	下降
壬二酸 Azelaic acid	C₉H₁₆O₄	1.12	0.96	0.012	下降
尿苷 Uridine	C₉H₁₂N₂O₆	1.35	0.95	0.001	下降
视黄酯 Retinyl ester	C₂₀H₃₀O₂	1.24	0.96	0.004	下降
顺-9,10-环氧硬脂酸 cis-9,10-Epoxystearic acid	C₁₈H₃₄O₃	1.66	0.92	0.025	下降
异柠檬酸盐 Isocitrate	C₆H₈O₇	1.55	1.12	0.047	上升
戊二酸 Glutaric acid	C₅H₈O₄	2.53	1.38	0.015	上升
6-脱氧噬菌体胺 (6-脱氧花青) 6-Deoxyfagomine	C₆H₁₃NO₂	1.36	0.94	0.012	下降
乙酰-L-酪氨酸 Acetyl-L-tyrosine	C₁₁H₁₃NO₄	1.38	0.87	0.010	下降
γ-谷氨酰鸟氨酸 Gamma glutamyl ornithine	C₁₀H₁₉N₃O₅	1.87	1.58	0.023	上升
对茴香酸异戊酯 (异戊基异茴香酸酯) Isoamyl p-anisate	C₁₃H₁₈O₃	1.76	0.67	0.041	下降
3-氧十二酸 3-Oxododecanoic acid	C₁₂H₂₂O₃	1.01	0.92	0.043	下降
C-2神经酰胺 C-2 Ceramide	C₂₀H₃₉NO₃	2.21	0.75	0.028	下降
磷脂酰乙醇胺 PE(15:0/16:1(9Z))	C₃₆H₇₀NO₈P	1.42	1.10	0.030	上升
半乳糖神经酰胺 Galactosyl ceramide (d18:1/14:0)	C₃₈H₇₃NO₈	1.97	2.78	0.045	上升
N-棕榈酰甘氨酸 N-Palmitoyl glycine	C₁₈H₃₅NO₃	1.97	0.86	0.000	下降
二十碳五烯酸 Eicosapentaenoic acid	C₂₀H₃₀O₂	1.01	0.98	0.004	下降
溶血磷脂酰乙醇胺 LysoPE(0:0/20:2(11Z,14Z))	C₂₅H₄₈NO₇P	1.33	1.12	0.037	上升
9-羟基癸酸 9-Hydroxydecanoic acid	C₁₀H₂₀O₃	1.21	0.92	0.028	下降
甘油一脂 MG(a-13:0/0:0/0:0)[rac]	C₁₆H₃₂O₄	1.92	0.79	0.040	下降
反式-2-十二碳烯二酸 Traumatic acid	C₁₂H₂₀O₄	1.37	0.86	0.025	下降
9-氧壬酸 9-Oxo-nonanoic acid	C₉H₁₆O₃	1.15	0.93	0.019	下降
8-羟基-5,6-辛二烯酸 8-Hydroxy-5,6-octadienoic acid	C₈H₁₂O₃	1.04	0.94	0.016	下降
(S)-3-磺酸盐 (S)-3-Sulfonatolactate	C₃H₆O₆S	2.57	3.05	0.046	上升
γ-谷氨酰缬氨酸 gamma-Glutamylvaline	C₁₀H₁₈N₂O₅	1.80	0.87	0.008	下降
穗花牡荆苷 Agnuside	C₂₂H₂₆O₁₁	1.41	0.91	0.005	下降
假尿苷 Pseudouridine	C₉H₁₂N₂O₆	1.19	0.93	0.003	下降
雌三醇7-(6-反式-对-香豆酰基葡萄糖苷) Eriodictyol 7-(6-trans-p-coumaroylglucoside)	C₃₀H₂₈O₁₃	1.44	0.92	0.020	下降
注：FC表示某差异代谢物在淡水组相对于海水组的表达倍数变化，FC>1表示该代谢物上调，FC<1表示该代谢物下调；P<0.05表示差异显著，P<0.01表示差异极显著。 Note: FC value indicates the change of expression multiple of a differential metabolite in FOV group compared with SOV group. FC>1 indicates that the metabolite is upregulated; FC<1 indicates that the metabolite is down-regulated. P<0.05 indicates significant difference, and P<0.01 indicates extremely significant difference.

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表 2 差异代谢产物的KEGG通路富集表

Table 2. KEGG pathway enrichment table of differential metabolites

富集的差异代谢产物名称　 Enriched differential metabolite name	富集个数 Numble	富集通路 ID Pathway	通路描述　　 Pathway desciption	P
半乳糖神经酰胺 Galactosyl ceramide (d18:1/14:0)	1	map00600	鞘脂代谢	0.059
脯氨酸 L-Proline	1	map00330	精氨酸和脯氨酸代谢	0.176
异柠檬酸盐 Isocitrate	1	map00020	柠檬酸循环	0.057
戊二酸 Glutaric acid	1	map00310	赖氨酸降解	0.130
皮质醇 Cortisol	1	map00140	类固醇激素生物合成	0.207
尿苷 Uridine；假尿苷 Pseudo uridine	2	map00240	嘧啶代谢	0.011
异柠檬酸盐 Isocitrate	1	map00630	乙醛酸和二羧酸代谢	0.137
L-异亮氨酸 L-Isoleucine；脯氨酸 L-Proline	2	map00970	氨酰tRNA生物合成	0.008
L-异亮氨酸 L-Isoleucine	1	map00290	缬氨酸、亮氨酸和异亮氨酸生物合成	0.064
二十碳五烯酸 Eicosapentaenoic acid	1	map01040	不饱和脂肪酸生物合成	0.097
L-异亮氨酸 L-Isoleucine	1	map00280	缬氨酸、亮氨酸和异亮氨酸降解	0.107
(S)-3-磺酸盐 (S)-3-Sulfonatolactate	1	map00270	半膀氨酸和蛋氨酸代谢	0.141

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