代乳粉添加单宁酸对7—28日龄湖羊羔羊胃肠道发育的影响

doi:10.3864/j.issn.0578-1752.2019.21.019

中国农业科学 ›› 2019, Vol. 52 ›› Issue (21): 3924-3933.doi: 10.3864/j.issn.0578-1752.2019.21.019

• 畜牧·兽医·资源昆虫 • 上一篇下一篇

代乳粉添加单宁酸对7—28日龄湖羊羔羊胃肠道发育的影响

郑琛¹,李发弟^2,³,李飞²,周巨旺¹,段鹏伟¹,刘绘汇¹,樊海苗¹,朱威力¹,刘婷¹()

1 甘肃农业大学动物科学技术学院,兰州 730070
2 兰州大学草地农业生态系统国家重点实验室/兰州大学农业农村部草牧业创新重点实验室/ 兰州大学草地农业科技学院,兰州 730020
3 甘肃省肉羊繁育生物技术工程实验室,甘肃民勤 733300

收稿日期:2019-06-24 接受日期:2019-08-06 出版日期:2019-11-01 发布日期:2019-11-12
作者简介:郑琛,E-mail： zhengc@gsau.edu.cn。
基金资助:
甘肃农业大学青年研究生指导教师扶持基金项目(GAU-QNDS-201603)

Effects of Tannic Acid Addition in Milk Replacer on Development of Gastrointestinal Tract of 7 to 28 Days Old Hu Lambs

ZHENG Chen¹,LI FaDi^2,³,LI Fei²,ZHOU JuWang¹,DUAN PengWei¹,LIU HuiHui¹,FAN HaiMiao¹,ZHU WeiLi¹,LIU Ting¹()

1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070
2 State Key Laboratory of Grassland Agro-ecosystems, Lanzhou University/Key laboratory of Grassland Livestock Industry Innovation, Ministry of Agriculture and Rural Affairs, Lanzhou University/College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730020
3 Engineering Laboratory of Mutton Sheep Breeding and Reproduction Biotechnology in Gansu Province, Minqin 733300, Gansu

Received:2019-06-24 Accepted:2019-08-06 Online:2019-11-01 Published:2019-11-12

摘要/Abstract

摘要：

目的观察代乳粉中添加单宁酸对7—28日龄湖羊羔羊胃肠道生长发育的影响。方法选择同质性好的7日龄湖羊公羔（双羔）30只,随机分为2组,每组15只,每只1个重复,分别饲喂对照代乳粉或单宁酸含量为0.2%的代乳粉,试验期21 d。羔羊28日龄时饲养试验结束,每组随机选择8只羔羊进行屠宰,分离胃室和肠段,分别称量各胃室和各肠段净质量以及含内容物质量,并量取各肠段的长度,计算各胃室和各肠段的相对质量和相对长度及内容物分布。肠道组织形态学和上皮细胞凋亡率用皱胃胃底腺区及十二指肠、空肠和回肠中段的组织样品测定。肠道屏障功能相关的闭锁蛋白（occludin）、闭锁小带1（zonula occludens-1,ZO-1）和紧密连接蛋白1（claudin 1）mRNA表达量用采集的十二指肠、空肠和回肠黏膜测定。结果除十二指肠指数（%活体质量,P=0.012）和相对质量（%全肠质量,P=0.034;%全胃肠质量,P=0.017）、空肠和结肠相对长度（%全肠长度,P=0.030,P=0.004）及结肠内容物分布（%活体质量,P=0.039）外,单宁酸对羔羊胃肠指数（%活体质量）、胃肠相对质量（%全胃质量、%全肠质量和%全胃肠质量）、肠道相对长度（%全肠长度）、内容物相对质量（%活体质量）、胃肠内容物相对总胃/肠内容物及总胃肠内容物相对质量（%总胃内容物、总肠内容物、总胃肠内容物）以及小肠claudin 1蛋白mRNA表达量均没有产生显著影响（P＞0.05）,但显著提高羔羊十二指肠肌层厚度并降低绒毛宽度（P=0.013,P=0.001）,显著上调空肠ZO-1蛋白mRNA表达量（P=0.003）,此外,单宁酸有上调羔羊空肠occludin蛋白mRNA表达量并降低空肠绒毛宽度和隐窝深度以及空肠和回肠上皮细胞凋亡率的趋势（P=0.077,P=0.073,P=0.062,P=0.097,P=0.052）。结论单宁酸显著降低7—28日龄湖羊羔羊十二指肠相对质量和空肠相对长度,但可通过提高十二指肠肌层厚度、上调空肠ZO-1蛋白和occludin蛋白mRNA表达量并降低空肠隐窝深度及空肠和回肠上皮细胞凋亡率来改善肠屏障功能。

关键词: 单宁酸, 羔羊, 代乳粉, 胃肠道, 发育

Abstract:

【Objective】 This study was conducted to investigate the effects of tannic acid supplementation to milk replacer on the development of gastrointestinal tract of 7 to 28 day-old Hu lambs. 【Method】 Thirty 7 day-old Hu male lambs were chosen and divided into 2 groups, fifteen lambs in each group and each lamb as a duplication. Lambs were fed milk replacer with or without 0.2% tannic acid, respectively. The test lasted 21 days. Eight lambs were selected from each group randomly and slaughtered at 28 day-old. The weights of the compound stomach and the intestinal tract with and without content, and lengths of the intestinal tract were measured, then the relative quality and length were calculated. While the tissue samples from fundus gland region of the abomasum, the middle part of duodenum, jejunum and ileum were fixed in paraformaldehyde to analyse the histomorphology, and the apoptotic rate of intestinal epithelial cells as well. And the mRNA expression of occludin, ZO-1 and claudin 1 protein of duodenum, jejunum and ileum mucosa were measured.【Result】 The results showed that except relative weights of duodenum (% body weight, P=0.012; % intestinal tract weight, P=0.034; % gastrointestinal tract weight, P=0.017), relative length of jejunum and colon (% intestinal tract length, P=0.030, P=0.004), and content of colon (% body weight, P=0.039), the relative weights (% body weight, % stomach weight, % intestinal tract weight, and % gastrointestinal tract weight), relative lengths (% intestinal tract length), content of stomach and intestinal tract (% body weight, % stomach content weight, % intestinal tract content weight, and % gastrointestinal tract content weight), and mRNA expression of claudin 1 protein in intestinal tract of lambs were not affected by tannic acid (P＞0.05). However, tannic acid elongated the muscular thickness of duodenum and decreased the villus width of lamb duodenum significantly (P=0.013,P=0.001), and up-regulated mRNA expression of ZO-1 protein of jejunum (P=0.003). And there was a tendency that tannic acid up-regulated mRNA expression of occludin protein of jejunum (P=0.077), and decreased villus width and crypt depth of jejunum and the apoptotic rate of jejunum and ileum cells (P=0.073, P=0.062, P=0.097, P=0.052). 【Conclusion】 In conclusion, tannic acid decreased the relative weights of duodenum and relative length of jejunum of 7-28 day-old Hu lambs, but improved the barrier function of intestinal tract via elongating the muscular thickness of duodenum, up-regulating the mRNA expression of ZO-1 and occludin protein of jejunum, and decreasing the crypt depth of jejunum and the apoptotic rate of jejunum and ileum cells.

Key words: tannic acid, lamb, milk replacer, gastrointestinal tract, development

郑琛,李发弟,李飞,周巨旺,段鹏伟,刘绘汇,樊海苗,朱威力,刘婷. 代乳粉添加单宁酸对7—28日龄湖羊羔羊胃肠道发育的影响[J]. 中国农业科学, 2019, 52(21): 3924-3933.

ZHENG Chen,LI FaDi,LI Fei,ZHOU JuWang,DUAN PengWei,LIU HuiHui,FAN HaiMiao,ZHU WeiLi,LIU Ting. Effects of Tannic Acid Addition in Milk Replacer on Development of Gastrointestinal Tract of 7 to 28 Days Old Hu Lambs[J]. Scientia Agricultura Sinica, 2019, 52(21): 3924-3933.

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图/表 6

表1

项目 Item		对照组 Control	单宁酸组 Tannic acid	SEM	P值 P value
胃室 Stomach
瘤胃 Rumen	%活体质量 % body weight	0.54	0.55	0.02	0.856
	%全胃质量 % the quality of total stomach	29.70	31.03	1.10	0.564
	%全胃肠质量 % the quality of total gastrointestinal tract	6.69	7.50	0.34	0.247
网胃 Reticulum	%活体质量 % body weight	0.16	0.16	0.01	0.864
	%全胃质量 % the quality of total stomach	9.16	9.03	0.35	0.859
	%全胃肠质量 % the quality of total gastrointestinal tract	2.04	2.19	0.09	0.426
瓣胃 Omasum	%活体质量 % body weight	0.08	0.08	0.01	0.659
	%全胃质量 % the quality of total stomach	4.08	4.59	0.32	0.450
	%全胃肠质量 % the quality of total gastrointestinal tract	0.92	1.14	0.10	0.304
皱胃 Abomasum	%活体质量 % body weight	1.04	1.00	0.05	0.683
	%全胃质量 % the quality of total stomach	57.05	55.34	1.23	0.506
	%全胃肠质量 % the quality of total gastrointestinal tract	12.77	13.40	0.40	0.448
肠道 Intestinal tract
十二指肠 Duodenum	%活体质量 % body weight	0.20^a	0.15^b	0.01	0.012
	%全肠质量 % the quality of total intestinal tract	3.21^a	2.69^b	0.12	0.034
	%全胃肠质量 % the quality of total gastrointestinal tract	2.48^a	2.04^b	0.10	0.017
空肠Jejunum	%活体质量 % body weight	4.00	3.59	0.15	0.162
	%全肠质量 % the quality of total intestinal tract	63.93	63.29	0.73	0.675
	%全胃肠质量 % the quality of total gastrointestinal tract	49.63	47.98	0.79	0.317
回肠 Ileum	%活体质量 % body weight	0.15	0.14	0.01	0.878
	%全肠质量 % the quality of total intestinal tract	2.35	2.52	0.17	0.642
	%全胃肠质量 % the quality of total gastrointestinal tract	1.83	1.91	0.14	0.772
盲肠 Cecum	%活体质量 % body weight	0.35	0.29	0.02	0.072
	%全肠质量 % the quality of total intestinal tract	5.54	5.16	0.25	0.456
	%全胃肠质量 % the quality of total gastrointestinal tract	4.30	3.90	0.19	0.321
结肠 Colon	%活体质量 % body weight	1.18	1.11	0.05	0.522
	%全肠质量 % the quality of total intestinal tract	18.85	19.64	0.53	0.477
	%全胃肠质量 % the quality of total gastrointestinal tract	14.61	14.88	0.40	0.750
直肠 Rectum	%活体质量 % body weight	0.38	0.37	0.02	0.755
	%全肠质量 % the quality of total intestinal tract	6.11	6.70	0.34	0.407
	%全胃肠质量 % the quality of total gastrointestinal tract	4.73	5.06	0.24	0.517

表3

表4

单宁酸对羔羊胃肠内容物相对质量的影响"

项目 Item		对照组 Control	单宁酸组 Tannic acid	SEM	P值 P value
胃室内容物 Stomach content
瘤胃内容物 Rumen content	%活体质量 % body weight	1.44	1.75	0.32	0.648
	%全胃内容物质量 % the quality of total stomach content	26.96	37.02	4.58	0.287
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	17.32	22.73	3.15	0.410
网胃内容物 Reticulum content	%活体质量 % body weight	0.27	0.20	0.06	0.553
	%全胃内容物质量 % the quality of total stomach content	4.99	3.39	0.91	0.405
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	3.51	2.13	0.71	0.358
瓣胃内容物 Omasum content	%活体质量 % body weight	0.01	0.01	0.0005	0.776
	%全胃内容物质量 % the quality of total stomach content	0.23	0.31	0.03	0.191
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	0.14	0.17	0.01	0.333
皱胃内容物 Abomasum content	%活体质量 % body weight	3.43	2.27	0.30	0.053
	%全胃内容物质量 % the quality of total stomach content	68.58	60.65	4.65	0.412
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	43.32	34.89	3.00	0.168
肠道内容物 Intestinal tract content
十二指肠内容物 Duodenum content	%活体质量 % body weight	0.15	0.09	0.02	0.111
	%全肠内容物质量 % the quality of total intestinal tract content	7.25	3.46	1.44	0.198
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	1.99	1.47	0.29	0.392
空肠内容物 Jejunum content	%活体质量 % body weight	1.26	1.10	0.18	0.670
	%全肠内容物质量 % the quality of total intestinal tract content	39.64	44.17	2.75	0.429
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	14.33	18.16	1.91	0.332
回肠内容物 Ileum content	%活体质量 % body weight	0.08	0.04	0.02	0.218
	%全肠内容物质量 % the quality of total intestinal tract content	1.75	1.77	0.32	0.978
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	0.80	0.75	0.16	0.904
盲肠内容物 Cecum content	%活体质量 % body weight	0.43	0.33	0.08	0.554
	%全肠内容物质量 % the quality of total intestinal tract content	12.85	13.20	1.82	0.928
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	4.90	4.89	0.64	0.991
结肠内容物 Colon content	%活体质量 % body weight	1.08^a	0.57^b	0.13	0.039
	%全肠内容物质量 % the quality of total intestinal tract content	33.28	25.72	2.57	0.148
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	12.53	10.96	1.42	0.596
直肠内容物 Rectum content	%活体质量 % body weight	0.17	0.23	0.04	0.449
	%全肠内容物质量 % the quality of total intestinal tract content	6.10	11.68	2.09	0.192
	%全胃肠内容物质量 % the quality of total gastrointestinal tract content	2.08	4.70	0.87	0.137

表4

表5

表6

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营养成分 Ingredient	含量 Concentration
干物质 Dry matter	95.68
粗蛋白质 Crude protein	24.89
粗脂肪 Ether extract	17.50
粗纤维 Crude fiber	2.16
粗灰分 Crude ash	5.81
钙 Calcium	1.02
总磷 Total phosphorus	0.59

项目Item	对照组Control	单宁酸组 Tannic acid	SEM	P值 P value
皱胃 Abomasum
黏膜厚度 Mucosa thickness (μm)	333.36	343.82	8.54	0.605
肌层厚度 Muscular thickness (μm)	548.48	278.36	52.15	0.810
十二指肠 Duodenum
绒毛高度 Villus height (μm)	361.99	425.00	21.54	0.150
绒毛宽度 Villus width (μm)	80.04^a	54.80^b	4.43	0.001
隐窝深度 Crypt depth (μm)	148.87	156.30	8.10	0.665
肌层厚度 Muscular thickness (μm)	161.93^b	208.41^a	10.01	0.013
绒隐比 V / C	2.57	2.76	0.13	0.505
空肠Jejunum
绒毛高度 Villus height (μm)	431.59	378.90	19.85	0.194
绒毛宽度 Villus width (μm)	71.13	64.74	1.79	0.073
隐窝深度 Crypt depth (μm)	173.48	143.65	8.05	0.062
肌层厚度 Muscular thickness (μm)	157.73	146.10	7.41	0.451
绒隐比 V / C	2.59	2.79	0.07	0.154
回肠 Ileum
绒毛高度 Villus height (μm)	373.81	383.77	29.23	0.883
绒毛宽度 Villus width (μm)	68.91	78.12	2.76	0.108
隐窝深度 Crypt depth (μm)	176.96	207.92	11.78	0.228
肌层厚度 Muscular thickness (μm)	274.31	302.91	46.72	0.792
绒隐比 V / C	2.19	1.85	0.12	0.194

项目Item	对照组Control	单宁酸组 Tannic acid	SEM	P值 P value
十二指肠 Duodenum
Occludin	0.86	1.16	0.10	0.124
ZO-1	0.91	1.07	0.08	0.317
Claudin 1	0.91	0.94	0.09	0.850
细胞凋亡 Apoptosis (%)	11.27	7.90	2.97	0.595
空肠Jejunum
Occludin	0.82	1.39	0.16	0.077
ZO-1	0.71^b	1.29^a	0.11	0.003
Claudin 1	0.97	1.10	0.25	0.800
细胞凋亡 Apoptosis (%)	21.09	3.99	5.13	0.097
回肠 Ileum
Occludin	0.95	1.02	0.10	0.744
ZO-1	0.88	1.11	0.10	0.249
Claudin 1	0.80	1.00	0.16	0.544
细胞凋亡 Apoptosis (%)	47.29	20.76	7.00	0.052

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代乳粉添加单宁酸对7—28日龄湖羊羔羊胃肠道发育的影响

Effects of Tannic Acid Addition in Milk Replacer on Development of Gastrointestinal Tract of 7 to 28 Days Old Hu Lambs

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项目Item	对照组Control	单宁酸组 Tannic acid	SEM	P值 P value
十二指肠 Duodenum	2.28	2.08	0.11	0.361
空肠Jejunum	81.90^a	80.33^b	0.37	0.030
回肠 Ileum	0.84	0.84	0.06	0.990
盲肠 Cecum	0.90	0.93	0.05	0.848
结肠 Colon	11.84^b	13.56^a	0.33	0.004
直肠 Rectum	2.23	2.26	0.13	0.886