高糖饲料对草鱼生长及肝脏糖代谢的影响

陈志钊; 朱涛; 雷彩霞; 姜鹏; 杜金星; 朱俊杰; 宋红梅; 李胜杰

doi:10.12131/20230020

高糖饲料对草鱼生长及肝脏糖代谢的影响

doi: 10.12131/20230020

陈志钊^{1, 2,},
朱涛²,
雷彩霞²,
姜鹏²,
杜金星²,
朱俊杰¹,
宋红梅²,
李胜杰^2, ,

1.
湖州师范学院生命科学学院，浙江湖州 313000
2.
中国水产科学研究院珠江水产研究所/农业农村部热带亚热带水产资源利用与养殖重点实验室，广东广州 510380

基金项目: 国家重点研发计划项目 (2021YFD1200804)；中国水产科学研究院基本科研业务费专项资金 (2020TD34, 2021XT0301)；国家自然科学基金青年基金项目 (31902357)

详细信息

作者简介:
陈志钊 (1998—)，男，硕士研究生，研究方向为水产动物遗传育种。E-mail: Zhizhao0312@163.com

通讯作者:
李胜杰 (1981—)，男，研究员，博士，研究方向为水产生物技术与遗传育种。E-mail: ssjjli@163.com

中图分类号: S 963
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出版历程
- 收稿日期: 2023-02-15
- 修回日期: 2023-05-19
- 录用日期: 2023-05-25
- 网络出版日期: 2023-06-21
- 刊出日期: 2023-10-05

Effects on growth and hepatic glucose metabolism of grass carp fed with high dietary carbohydrates

1.
School of Life Science, Huzhou University, Huzhou 313000, China
2.
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences/ Key Laboratory of Tropical & Subtropical Fishery Resource Application & Cultivation, Ministry of Agriculture and Rural Affairs, Guangzhou 510380, China

摘要

摘要: 为了从生长性状、器官健康和营养代谢水平方面探究草食性鱼类草鱼 (Ctenopharyngodon idella) 对高糖饲料的适应和利用能力，以初始体质量 (132.01±16.43) g的草鱼为研究对象，分别配制正常糖水平 (15%，对照组) 和高糖水平 (40%，高糖组) 的等氮等能饲料分组投喂草鱼140 d，每组设3个重复，每个重复300 尾鱼。结果显示，相较于对照组，第60、第80 和第140天时高糖组草鱼的体质量显著增加 (P<0.05)，第140天时，高糖组草鱼的脏体比、肠系膜脂肪系数、中肠绒毛高度和肝糖原含量均显著提高且肝脏和中肠肠道组织出现损伤。随实验时间的延长，高糖组葡萄糖激酶 (gk)、丙酮酸激酶 (pk)、糖原合酶 (gys2)、脂肪酸合成酶 (fas) 基因表达量呈下降趋势，磷酸烯醇式丙酮酸激酶 (pepck)、葡萄糖-6-磷酸酶 (g6pase) 基因表达量呈上升趋势，对照组gys2、fas、pepck和g6pase基因表达量变化趋势与高糖组一致，但gk、pk基因表达量在各时间点均无显著性变化。第140天时，与对照组相比，高糖组pk基因表达量显著下调，g6pase、pepck和fas基因表达量显著上调。综上，采用高糖饲料投喂草鱼可促进其体质量增加，但长期饲喂会对其生理指标造成一定负面影响，肝脏糖代谢相关基因表达变化显著。
- 草鱼 /
- 糖代谢 /
- 肝脏 /
- 肠道结构
Abstract: In order to explore the growth performance, organic health and nutritional metabolism level of Ctenopharyngodon idella, a fed herbivorous fish-grass carp, with high dietary carbohydrates, we divided 1 800 individuals of (132.01±16.43) g into normal carbohydrate diet group (Control group) and high carbohydrate diet group (Group H), feeding them with 15% and 45% level carbohydrate diets for 140 d. Each group included three replications and each replication included 300 individuals. The results show that compared with the control group, the body mass of Group H was higher significantly on 60^th, 80^th and 140^th day (P<0.05). The visceral body ratio, mesenteric fat coefficient, villus height and hepatic glycogen content of Group H increased significantly on 140^th day, and damage was observed in liver and intestinal tract. The expression of glucose kinase gene (gk), pyruvate kinase gene (pk), glycogen synthase 2 gene (gys2) and fatty acid synthetase gene (fas) showed a decreasing trend in Group H along with time, while phosphoenolpyruvate carboxykinase gene (pepck) and glucose-6-phosphatase gene (g6pase) showed an increasing trend. The expressions of gys2, fas, pepck and g6pase had the same trend in Group L as that in Group H, while no significant variation trend was observed in gk and pk gene at each time. Compared with the control group, the expression of pk was significantly lower, while those of g6pase, pepck and fas were significantly higher in Group H. In conclusion, 40% level dietary carbohydrate can improve the weight gain rate of grass carp, but high dietary carbohydrates have a negative impact on physiological indicators and significantly affect glucose metabolism at molecular level.
- Ctenopharyngodon idella /
- Glucose metabolism /
- Liver /
- Intestinal structure

HTML全文

图 1 草鱼肝脏组织结构

注：a. 对照组草鱼肝脏 (400×)；b. 高糖组草鱼肝脏 (400×)；c. 对照组草鱼肝脏 (1 000×)；d. 高糖组草鱼肝脏 (1 000×)。

Figure 1. Tissue slices of liver of grass carp

Note: a. Liver of grass carp (400×); b. Liver in Group H of grass carp (400×); c. Liver in control group of grass carp (1 000×); d. Liver in Group H of grass carp (1 000×).

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图 2 草鱼中肠组织结构

注：a. 对照组草鱼中肠 (100×)；b. 高糖组草鱼中肠 (100×)；c. 对照组草鱼中肠 (400×)；d. 高糖组草鱼中肠 (400×)。

Figure 2. Tissue slices of midgut of grass carp

Note: a. Midgut in Control group grass carp (100 ×); b. Midgut in Group H of grass carp (100×); c. Midgut in control group of grass carp (400×); d. Midgut in Group H of grass carp (400×).

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图 3 高糖饲料对草鱼肠道组织绒毛高度、宽度和肌层厚度的影响

Figure 3. Effects of dietary carbohydrate on villus height, villus width and muscle layer thickness

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图 4 草鱼肝脏糖代谢相关基因的表达

注：方柱上的不同大写字母表示同一时间点高糖组、对照组的相对表达量差异显著，不同小写字母表示同一组在不同时间点的相对表达量差异显著 (P<0.05)。

Figure 4. Relative expression of genes related to liver glucose metabolism in grass carp

Note: Different uppercase letters on the square columns indicate significant differences in relative expression levels between the high glucose group and the control group at the same time point, while different lowercase letters on the square columns indicate significant differences in relative expression levels of the same group at different time points (P<0.05).

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表 1 实验饲料配方及常规成分分析

Table 1. Formulation and proximate composition of diets

主要成分 Essential component	普通饲料 Normal diet/%	高糖饲料 High-carbohydrate diet/%
鸡肉粉 Poultry meal	5	5
普通蒸汽级鱼粉 Ordinary fish meal	15	15
豆粕 Bean meal	33.57	33.57
木薯生淀粉 Raw cassava starch	15	40
豆油 Soybean oil	3	3
细统糠 Fine bran	25	0
多维多矿预混料^① Premix (1%)	1	1
磷酸二氢钙 Ca(H₂PO₄)₂	2	2
氯化胆碱 Choline chloride (50%)	0.2	0.2
食盐 Common salt	0.2	0.2
抗氧化剂 Antioxidant	0.03	0.03
合计 Total	100	100
营养成分 Nutritional ingredient
粗蛋白 Primary protein	28.55	28.55
粗脂肪 Crude fat	5.22	5.32
钙 Ca	1.35	1.35
磷 P	1.41	1.41
赖氨酸 Lys	1.83	1.83
蛋氨酸 Met	0.53	0.53
注：① 多维多矿预混料为商用预混料，由淮安市禾丰饲料有限公司提供。 Note: ① Multi-mineral premix is a commercial premix provided by Huai'an Hefeng Feed Co. Ltd..

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表 2 草鱼糖代谢相关基因 qRT-PCR 引物

Table 2. qRT-PCR primers of grass carp glycometabolism genes

基因 Gene	引物序列 (5'—3') Primer sequence (5'–3')	GenBank 登录号 GenBank ID	参考文献 Reference
gk	F: GGACAATGGTGAACTGGAGGACTTC R: TAGCAGCACAAGACGCACAAGC	GU065314	[19]
pk	F: GCCGAGAAAGTCTTCATCGCACAG R: CGTCCAGAACCGCATTAGCCAC	JQ951928	[20]
pepck	F: ATCGTCACGGAGAACCAA R: CCTGAACACCAAACTTAGCA	JQ898294.1	[20]
g6pase	F: AAGGACAGCAGGTAGAAGAGG R: ACGGAAAACAAGAAGAGCAG	KY742725.1
gys2	F: CCTCCAGTAACAACTCACAACA R: CAGATAGATTGGTGGTTACGC	JQ792167	[19]
fas	F: GATGGGTCTAAGCCTGATGG R: GACACCCTGTGGACATTGAGC	MG241310	[19]
β-actin	F: GCTATGTGGCTCTTGACTTCG R: GGGCACCTGAACCTCTCATT	M25013.1	[21]

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表 3 高糖饲料对草鱼生长性能的影响

Table 3. Effects of dietary carbohydrate on growth performance of grass carp

项目 Item	样本量/组 N/Group	养殖时间 Culture time/d	对照组 Control group	高糖组 Group H
初始体质量 Initial body mass/g	60	0	130.61±14.79	132.83±13.36
终末体质量 Final body mass/g	60	40	146.08±27.97	143.88±20.43
		60	153.18±29.40	165.54±36.64^*
		80	168.71±36.73	195.53±48.70^*
		140	361.18±67.61	388.57±59.02^**
体质量增长率 R_F/%	60	40	109.98±15.64	110.18±9.33
		60	121.24±4.21	133.28±4.09^*
		80	129.82±0.23	147.20±9.68^*
		140	271.78±13.07	297.51±15.85
体长 Body length/cm	60	40	20.74±1.29	20.13±1.05
		60	20.34±1.21	20.75±1.41
		80	21.21±1.71	21.58±2.16
		140	29.27±0.82	29.22±1.09
体高 Body height/cm	60	40	4.90±0.37	4.96±0.37
		60	4.88±0.44	5.20±0.51^**
		80	5.34±0.59	5.49±0.70
		140	6.54±0.15	6.87±0.32^**
肥满度 R_F	60	40	1.61±0.13	1.76±0.32^**
		60	1.78±0.28	1.85±0.23
		80	1.77±0.19	1.92±0.28^**
		140	1.48±0.13	1.72±0.17^**
肝体比 R_HI/%	30	40	1.94±0.47	2.19±0.62
		60	1.68±0.61	2.14±0.60
		80	2.17±0.27	2.29±0.49
		140	1.23±0.24	1.39±0.14
脏体比 R_VI/%	30	140	6.64±1.29	8.12±0.71^**
肠系膜脂肪系数 R_MFI/%	30	140	1.16±0.54	2.20±0.64^**
空壳质量 W_E/g	30	140	345.71±20.67	383.23±27.46^**
注：. 同一时期下高糖组与对照组相比差异显著 (P<0.05)；. 差异极显著 (P<0.01)。后表同此。 Note: . There are significant differences between the high glucose group and the control group during the same period (P<0.05); **. Extremely significant differences (P<0.01). The same case in the following table.

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表 4 高糖饲料对草鱼血糖、胰岛素、糖原的影响

Table 4. Effects of dietary carbohydrate on plasma glucose, insulin and glycogen of grass carp

项目　　　　　　 Item	养殖时间 Culture time/d	对照组 Control group	高糖组 Group H
血糖浓度 Blood glucose/(mmol·L⁻¹)	40	10.95±0.99	10.99±1.17
	60	11.46±2.15	13.65±1.63
	80	9.77±1.77	10.79±1.78
	140	13.39±0.63	12.78±1.00
胰岛素水平 Insulin level/(mIU·L⁻¹)	140	9.23±1.18	10.31±1.17
肝糖原质量分数 Liver glycogen/(mg·g⁻¹)	140	24.23±13.03	36.29±10.41^*
肌糖原质量分数 Muscle glycogen/(mg·g⁻¹)	140	0.15±0.06	0.24±0.08

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