单宁酸对免疫应激断奶仔猪肌肉形态、风味氨基酸及肌纤维相关基因表达的影响

doi:10.11843/j.issn.0366-6964.2025.03.028

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (3): 1290-1301.doi: 10.11843/j.issn.0366-6964.2025.03.028

单宁酸对免疫应激断奶仔猪肌肉形态、风味氨基酸及肌纤维相关基因表达的影响

周文涛¹^,²(), 王晨昱¹^,², 周辉¹^,², 刘洪彪³, 冯舒锾¹, 范高升¹, 李铁军²^,*(), 何流琴¹^,²^,*()

1. 湖南师范大学生命科学学院动物肠道功能调控湖南省重点实验室湖南省动物肠道生态与健康国际科技创新合作基地动物营养与人体健康实验室, 长沙 410081
2. 中国科学院亚热带农业生态研究所动物营养生理与代谢过程湖南省重点实验室, 长沙 410125
3. 张家界久瑞生物科技有限公司, 张家界 427099

收稿日期:2024-06-12 出版日期:2025-03-23 发布日期:2025-04-02
通讯作者: 李铁军,何流琴 E-mail:zhouwent213@163.com;tjli@isa.ac.cn;heliuqin@hunnu.edu.cn
作者简介:周文涛(1998-)，男，湖南衡阳人，硕士生，主要从事动物营养与饲料科学研究，E-mail：zhouwent213@163.com
基金资助:
国家自然科学基金(32172755；3210099)；国家自然科学区域创新联合基金重点支持项目(U23A20233)；湖南省科技创新领军人才项目(2023RC1054)；湖南省重点研发项目(2022NK2023)

Effects of Tannic Acid on Muscle Morphology, Flavor Amino Acids, and Expression of Muscle Fiber-related Genes in Immunostressed Weaned Piglets

ZHOU Wentao¹^,²(), WANG Chenyu¹^,², ZHOU Hui¹^,², LIU Hongbiao³, FENG Shuhuan¹, FAN Gaosheng¹, LI Tiejun²^,*(), HE Liuqin¹^,²^,*()

1. Laboratory of Animal Nutrition and Human Health, Hunan International Joint Laboratory of Animal Intestinal Ecology and Health, Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation, College of Life Sciences, Hunan Normal University, Changsha 410081, China
2. Hunan Provincial Key Laboratory of Animal Physiology and Metabolic Process, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China
3. Zhangjiajie Jiurui Biology & Chemistry Co., Ltd, Zhangjiajie 427099, China

Received:2024-06-12 Online:2025-03-23 Published:2025-04-02
Contact: LI Tiejun, HE Liuqin E-mail:zhouwent213@163.com;tjli@isa.ac.cn;heliuqin@hunnu.edu.cn

摘要/Abstract

摘要：

旨在研究低蛋白质饲粮中添加单宁酸(TA)对大肠杆菌(ETEC)诱导的免疫应激断奶仔猪血液生化指标、肌肉形态结构、风味氨基酸及肌纤维发育相关因子的影响。选取体重为6.75 kg±0.07 kg的杜×长×大三元健康断奶仔猪21头，随机分为3组，分别为基础饲粮组(CON组)、基础饲粮+ETEC组(EK组)和基础饲粮+0.05% TA+ETEC组(MTA组)，每组7个重复，每个重复1头猪。在试验第29和31天给予EK组和MTA组断奶仔猪灌服20 mL ETEC浓度为1×10¹² CFU·mL^-1的LB培养基，CON组灌服同体积灭活的LB培养基，第32天采集血液和背最长肌样品用于检测分析。结果表明：1)不同处理对断奶仔猪的全血细胞数量和比例没有显著影响(P>0.05)。2)在血清生化参数中，ETEC处理可显著降低血清中高密度脂蛋白的含量，提高总胆汁酸含量，而日粮补充TA可逆转这些指标的水平，且MTA组的肌酐含量较其他两组均显著降低。3)ETEC处理可损伤背最长肌的形态结构，尤其是可显著降低肌纤维平均直径和面积，而饲粮中添加TA可有效维持其形态结构的完整性。4)CON组和MTA组背最长肌MyoD和MSTN的mRNA相对表达量与EK组相比均显著降低(P < 0.05)，EK组MyoG的mRNA相对表达量显著高于CON组(P < 0.05)，而MTA组的MyHC Ⅱ b mRNA丰度显著低于CON组和EK组(P < 0.05)。5)ETEC处理可使背最长肌中赖氨酸、亮氨酸、蛋氨酸、必需氨基酸、甜味氨基酸和总氨基酸含量均显著减少(P < 0.05)，但饲喂TA日粮可在一定程度上提高这些氨基酸的含量。综上所述，低蛋白质饲粮添加0.05% TA可通过维持断奶仔猪肌肉形态结构的完整性，调控肌纤维相关基因表达和风味氨基酸的含量来促进免疫应激断奶仔猪的肌肉发育。

关键词: 单宁酸, 断奶仔猪, 应激损伤, 肌纤维, 风味氨基酸

Abstract:

The objective of this experiment was to investigate the impact of tannic acid (TA) supplementation in low protein diets on blood biochemical parameters, muscle morphology and structure, flavor amino acids, and factors associated with myofibril development in weaned piglets experiencing immune stress induced by Escherichia coli (ETEC). Twenty-one healthy weaned piglets (Duroc×Landrace×Yorkshire), weighing approximately 6.75±0.07 kg, were selected and randomly allocated into three groups: basal diet group (CON group), basal diet+ETEC group (EK group) and basal diet+0.05% TA+ETEC group (MTA group). Each group consisted of seven replications, with one pig per replication. Weaned piglets in the EK and MTA groups were orally administered 20 mL of LB medium containing an ETEC concentration of 1×10¹² CFU·mL^-1 on days 29 and 31 of the experiment, while the CON group received the same volume of inactivated LB medium. On day 32, blood samples and longissimus dorsi were collected for assay analysis. The results were showed as follows: 1) The various treatments did not yield any statistically significant impact on the quantity and proportion of whole blood cells in weaned piglets (P>0.05). 2) Among the serum biochemical parameters, ETEC treatment significantly decreased the serum high-density lipoprotein (HDL) level and increased the total bile acid level, whereas TA treatment reversed these indices, and creatinine levels were significantly lower in the MTA group compared to both other groups. 3) ETEC treatment may induce alterations in the morphology and structure of the longissimus dorsi, particularly leading to a reduction in both average diameter and area of muscle fibers. However, supplementation of TA to a low protein diet effectively preserves the integrity of its morphology and structure. 4) The relative mRNA expression levels of MyoD and MSTN in the longissimus dorsi were significantly reduced in both the CON and MTA groups compared to the EK group (P < 0.05). Additionally, the relative mRNA expression level of MyoG was significantly higher in the EK group than in the CON group (P < 0.05), while there was a significant decrease in MyHC Ⅱ b mRNA abundance observed in the MTA group compared to both the CON and EK groups (P < 0.05). 5) The ETEC treatment significantly decreased the levels of lysine, leucine, methionine, essential amino acids, sweet amino acids, and total amino acids in longissimus dorsi (P < 0.05). However, feeding TA diets partially restored the content of the amino acids mentioned above. In conclusion, the inclusion of 0.05% TA in low-protein diets can enhance muscle development in immunostressed weaned piglets by preserving muscle morphology and structure integrity, as well as modulating the expression of myofiber-related genes and flavor amino acid content.

Key words: tannic acid, weaned piglets, stress injury, muscle fibers, flavor amino acids

中图分类号:

S828.5

周文涛, 王晨昱, 周辉, 刘洪彪, 冯舒锾, 范高升, 李铁军, 何流琴. 单宁酸对免疫应激断奶仔猪肌肉形态、风味氨基酸及肌纤维相关基因表达的影响[J]. 畜牧兽医学报, 2025, 56(3): 1290-1301.

ZHOU Wentao, WANG Chenyu, ZHOU Hui, LIU Hongbiao, FENG Shuhuan, FAN Gaosheng, LI Tiejun, HE Liuqin. Effects of Tannic Acid on Muscle Morphology, Flavor Amino Acids, and Expression of Muscle Fiber-related Genes in Immunostressed Weaned Piglets[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1290-1301.

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项目 Item	含量 Content	营养水平 Nutrient Level²	含量 Content
玉米 Corn	28.63	干物质 Dry matter	87.95
熟化大米 Ripening rice	26.40	消化能 Digestive energy	14.42
面粉 Flour	13.00	总能/(MJ·kg^-1) Gross energy	17.35
膨化大豆 Expanded soybean	6.00	粗蛋白质 Crude protein	16.27
豆粕(蛋白含量46%)Soybean meal	4.50	粗脂肪 Ether extract	4.51
发酵豆粕 Fermented soybean meal	4.50	钙 Calcium	0.80
蔗糖 Sucrose	3.00	总磷 Phosphorus	0.56
麦麸 Wheat bran	2.50	赖氨酸 Lysine	1.31
鱼粉(蛋白含量65%)Fish meal	3.00	甲硫氨酸+半胱氨酸 Methionine+Cystine	0.75
葡萄糖 Glucose	2.60	色氨酸 Tryptophan	0.23
大豆油 Soybean oil	1.70	苏氨酸 Threonine	0.79
磷酸氢钙 CaHPO₄	1.15
石粉 Limestone	0.90
赖氨酸盐酸盐 lysine hydrochloride	0.77
氯化钠 NaCl	0.40
苏氨酸 Threonine	0.28
甲硫氨酸 Methionine	0.22
L-缬氨酸 L-Valine	0.10
L-色氨酸 L-Tryptophan	0.06
氯化胆碱 Choline chloride (50%)	0.10
预混料 Premix¹	0.19
合计 Total	100.00

基因 Gene	序列号 Accession number	引物序列(5'→3') Primer sequence(5'→3')	产物长度/bp Product length
β-肌动蛋白 β-actin	XM_003124280.3	F: CTGCGGCATCCACGAAACT R: AGGGCCGTGATCTCCTTCTG	147
肌球蛋白重链Ⅰ MyHC Ⅰ	NM_213855.2	F: AAGGGCTTGAACGAGGAGTAGA R: TTATTCTGCTTCCTCCAAAGGG	115
肌球蛋白重链Ⅱ a MyHC Ⅱ a	XM_021066217.1	F: TATCCACCAGCCTGAACCTT R: CTTGTGTCCCCGCATAATTCTC	140
肌球蛋白重链Ⅱ b MyHC Ⅱ b	XM_021066035.1	F: AGATGCCTCTGTCCTGTCCTT R: TAGCCATTTCCTGGTCGGAAC	151
肌球蛋白重链Ⅱ x MyHC Ⅱ x	XM_021066024.1	F: ACAAGTTGCGGGTCAAGAGT R: TTGTGTGCATTTCTTTGGTCAC	97
生肌决定因子 MyoD	NM_001002824.1	F: GAGAGCACTACAGCGGTGAC R: GCTGTAATAGGTGCCGTCGT	128
肌细胞生成素 MyoG	NM_001012406.1	F: CAGCGCCATCCAGTACATC R: GGTGAGGGAGTGCAGATTGT	238
肌肉生长抑制素 MSTN	NM_214435.2	F: CCCGTCAAGACTCCTACAACA R: GCCTGGGTTCATGTCAAGTT	114
肌源性因子5 Myf5	NM_001278775.1	F: CTGTCCGCAGAAGATGGACC R: CTCAAACTCGTCCCCGAACT	112

项目 Item	对照组 Control group	EK组 EK group	MTA组 MTA group	P值 P-value
白细胞数/(10⁹·L^－1) WBC	18.66±2.34	19.09±1.83	17.15±1.03	0.742
中性粒细胞数/(10⁹·L^－1) Neu	5.95±0.83	5.52±1.16	4.99±0.54	0.748
中性粒细胞百分比/% Neu	32.42±3.80	29.78±4.53	29.56±3.73	0.858
淋巴细胞数/(10⁹·L^－1) Lym	11.74±1.92	11.22±0.788	10.53±0.76	0.802
淋巴细胞百分比/% Lym	62.22±4.44	63.60±4.51	65.60±4.42	0.866
单细胞数/(10⁹·L^－1) Mon	0.66±0.10	0.74±0.16	0.46±0.06	0.261
单细胞百分比/% Mon%	3.10±0.42	4.00±0.39	2.78±0.40	0.124
嗜酸性粒细胞数/(10⁹·L^－1) Eos	0.30±0.04	0.38±0.09	0.31±0.04	0.604
嗜酸性粒细胞百分比/% Eos%	1.62±0.20	2.45±0.57	1.81±0.38	0.398
红细胞计数/(10¹²·L^－1) RBC	6.13±0.36	6.16±0.23	5.80±0.23	0.597
血红蛋白浓度/(g·L^－1) HGB	106.80±4.73	106.40±3.38	105.16±3.84	0.954
红细胞压积/% HCT	29.78±1.61	29.18±0.95	29.08±1.30	0.924
平均红细胞体积/fL MCV	48.74±2.09	48.16±0.58	50.60±0.35	0.395
平均红细胞血红蛋白含量/pg MCH	17.52±0.60	17.90±0.23	17.94±0.09	0.698
平均红细胞血红蛋白浓度/(g·L^－1) MCHC	364.50±2.87	369.16±6.06	358.60±3.23	0.310
红细胞分布宽/fL RDW-SD	43.08±1.94	41.02±0.977	42.72±0.77	0.525
血小板计数/(10⁹·L^－1) PLT	464.25±34.13	435.40±48.89	584.20±37.69	0.058
平均血小板体积/fL MPV	9.27±0.06	8.78±0.45	8.76±0.29	0.539
降钙素原/% PCT	0.43±0.02	0.40±0.05	0.52±0.29	0.108

项目 Item	对照组 Control group	EK组 EK group	MTA组 MTA group	P值 P-value
肌酐/(μmol·L^－1) CREA	76.50±3.69^a	82.16±1.97^a	65.57±4.35^b	0.014
葡萄糖/(mmol·L^－1) GLU	6.12±0.28	6.10±0.62	4.65±0.54	0.107
甘油三酯/(mmol·L^－1L) TG	0.54±0.07	0.52±0.10	0.45±0.06	0.669
胆固醇/(mmol·L^－1) CHOL	2.31±0.13	2.26±0.12	2.08±0.07	0.374
低密度脂蛋白-胆固醇/(mmol·L^－1) LDL-C	1.20±0.05	1.13±0.09	1.20±0.11	0.816
高密度脂蛋白-胆固醇/(mmol·L^－1) HDL-C	1.08±0.08^a	0.83±0.06^b	0.98±0.03^ab	0.040
总胆汁酸/(μmol·L^－1) TBA	49.7±4.44^b	68.97±5.68^a	41.18±5.88^b	0.008
总胆红素/(g·L^－1) TBIL	1.16±0.15	0.95±0.18	1.44±0.16	0.173

项目 Item	对照组 Control group	EK组 EK group	MTA组 MTA group	P值 P-value
肌纤维直径/μm Muscle fiber diameter	29.82±0.30^a	28.25±0.46^b	29.10±0.27^ab	0.030
肌纤维面积/mm² Muscle fiber area	737.37±13.76^a	646.60±21.26^b	701.19±12.64^a	0.007

单宁酸对免疫应激断奶仔猪肌肉形态、风味氨基酸及肌纤维相关基因表达的影响

Effects of Tannic Acid on Muscle Morphology, Flavor Amino Acids, and Expression of Muscle Fiber-related Genes in Immunostressed Weaned Piglets

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项目 Item	对照组 Control group	EK组 EK group	MTA组 MTA group	P值 P-value
肌球蛋白重链Ⅰ MyHC Ⅰ	1.00±0.19	0.85±0.08	0.89±0.13	0.793
肌球蛋白重链Ⅱ a MyHC Ⅱ a	1.00±0.12	1.46±0.14	1.17±0.28	0.289
肌球蛋白重链Ⅱ b MyHC Ⅱ b	1.00±0.04^a	0.98±0.09^a	0.62±0.05^b	0.003
肌球蛋白重链Ⅱ x MyHC Ⅱ x	1.00±0.04	0.94±0.03	0.97±0.09	0.839
生肌决定因子 MyoD	1.00±0.08^b	1.56±0.11^a	1.11±0.07^b	0.003
肌细胞生成素 MyoG	1.00±0.13^b	1.52±0.12^a	1.20±0.09^ab	0.022
肌肉生长抑制素 MSTN	1.00±0.10^b	1.45±0.16^a	0.87±0.16^b	0.034
肌源性因子5 Myf5	1.00±0.06	0.96±0.10	0.89±0.08	0.741

项目 Item	对照组 Control group	EK组 EK group	MTA组 MTA group	P值 P-value
谷氨酸 Glu	11.47±0.28	11.12±0.22	11.62±0.19	0.387
天冬氨酸 Asp	6.99±0.22	6.73±0.11	7.03±0.08	0.433
赖氨酸 Lys	7.04±0.17^a	6.57±0.08^b	6.87±0.06^ab	0.048
丙氨酸 Ala	4.39±0.14	4.07±0.10	4.35±0.09	0.118
苏氨酸 Thr	3.43±0.09	3.33±0.06	3.48±0.04	0.443
甘氨酸 Gly	3.72±0.12^ab	3.46±0.08^b	3.89±0.04^a	0.019
脯氨酸 Pro	3.07±0.12	2.91±0.05	3.18±0.02	0.170
丝氨酸 Ser	2.78±0.06	2.59±0.06	2.74±0.04	0.098
亮氨酸 Leu	6.38±0.16^a	5.89±0.09^b	6.15±0.07^ab	0.036
精氨酸 Arg	5.09±0.14	4.76±0.11	5.11±0.06	0.071
缬氨酸 Val	3.99±0.09	3.73±0.06	3.92±0.04	0.059
异亮氨酸 Ile	3.64±0.09	3.37±0.05	3.54±0.05	0.064
苯丙氨酸 Phe	3.62±0.17^a	2.91±0.05^b	3.11±0.68^b	0.001
酪氨酸 Tyr	2.85±0.13^a	2.24±0.03^b	2.42±0.04^b	<0.001
组氨酸 His	2.20±0.04	1.98±0.14	2.11±0.04	0.285
蛋氨酸 Met	1.58±0.06^a	1.37±0.02^b	1.56±0.04^a	0.018
必需氨基酸 EAA	29.92±0.81^a	27.28±0.43^b	28.65±0.33^ab	0.022
鲜味氨基酸 UAA	18.46±0.51	17.85±0.34	18.66±0.26	0.394
甜味氨基酸 SAA	24.97±0.69^a	23.04±0.55^b	24.62±0.18^ab	0.046
总氨基酸 TAA	73.30±1.97^a	66.82±1.62^b	71.24±0.76^ab	0.029