加味枳术散对断奶仔兔肠黏膜屏障和抗氧化能力的影响

doi:10.11843/j.issn.0366-6964.2024.12.044

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (12): 5825-5838.doi: 10.11843/j.issn.0366-6964.2024.12.044

加味枳术散对断奶仔兔肠黏膜屏障和抗氧化能力的影响

靳茹文¹^,²(), 王英杰¹(), 蒋全兴¹(), 刘天强³, 邓阳⁴, 罗杰⁵, 赵玲¹, 叶刚¹, 施飞¹, 李英伦¹, 唐华侨¹^,*()

1. 四川农业大学动物医学院, 成都 611130
2. 四川省川龙动科药业有限公司, 成都 611230
3. 通威农业发展有限公司, 成都 610041
4. 成都市畜禽遗传资源保护中心, 成都 610081
5. 铜仁职业技术学院, 铜仁 554300

收稿日期:2023-03-02 出版日期:2024-12-23 发布日期:2024-12-27
通讯作者: 唐华侨 E-mail:jingruwen123@163.com;1764720963@qq.com;jiangquanxing27@163.com;turtletang@163.com
作者简介:靳茹文(1987-), 男, 四川达州人, 硕士, 主要从事兽用中药对畜禽疾病防治研究和肠靶向制剂开发, E-mail: jingruwen123@163.com
王英杰(1997-), 女, 河南淮阳人, 硕士生, 主要从事兽医药理学和毒理学研究, E-mail: 1764720963@qq.com
蒋全兴(1998-), 男, 四川德阳人, 硕士生, 主要从事天然药物活性研究, E-mail: jiangquanxing27@163.com
第一联系人：
靳茹文、王英杰和蒋全兴为同等贡献作者

Effects of Modified Zhizhu Powder on Intestinal Mucosal Barrier and Antioxidant Capacity of Weaned Rabbits

JIN Ruwen¹^,²(), WANG Yingjie¹(), JIANG Quanxing¹(), LIU Tianqiang³, DENG Yang⁴, LUO Jie⁵, ZHAO Ling¹, YE Gang¹, SHI Fei¹, LI Yinglun¹, TANG Huaqiao¹^,*()

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2. Sichuan Chuanlong Dongke Pharmaceutical Co., Ltd., Chengdu 611230, China
3. Tongwei Agricultural Development Co., Ltd., Chengdu 610041, China
4. Chengdu Animal Genetic Resources Protection Center, Chengdu 610081, China
5. Tongren Polytechnic College, Tongren 554300, China

Received:2023-03-02 Online:2024-12-23 Published:2024-12-27
Contact: TANG Huaqiao E-mail:jingruwen123@163.com;1764720963@qq.com;jiangquanxing27@163.com;turtletang@163.com

摘要/Abstract

摘要：

旨在探讨加味枳术散对断奶仔兔回肠肠黏膜屏障的影响，为仔兔腹泻疾病的防治提供新思路。试验选取90只35日龄，体重相近的新西兰白兔，按照雌雄各半的原则随机分为5组。其中空白组仔兔饲喂基础日粮，恩诺沙星组、低剂量组、中剂量组和高剂量组分别在基础日粮中添加0.2%的恩诺沙星粉剂以及0.1%、0.3%、0.5%的加味枳术散粉剂，预饲期7 d，正式试验14 d。试验结束后检测仔兔的肠道屏障完整情况及其抗氧化能力。结果显示，加味枳术散中、高剂量组可以显著降低肠道的病理组织评分(P < 0.01)。与空白组相比，其他处理组均显著升高回肠ZO2(zonula occludens proteins 2)的mRNA表达量(P < 0.001)；高剂量组显著增加了Occludin、Claudin、ZO1(zonula occludens proteins-1)、ZO2以及JAM2 mRNA的水平(P < 0.05)；加味枳术散显著增加回肠MUC2蛋白的表达水平(P < 0.05)，且高剂量组显著降低MPO活性(P < 0.01)；加味枳术散高剂量组可显著增强回肠sIgA的分泌，降低促炎细胞因子IL-1β、IL-2、IL-6与 TNF-α的水平，提高抗炎细胞因子IL-10与TGF-β的水平；加味枳术散可提高回肠SOD、GSH、CAT的活性和T-AOC。综上，加味枳术散能够有效改善断奶仔兔回肠形态结构、加强肠道细胞间的紧密连接，维持肠黏膜通透性，降低肠道的炎症反应，增强肠黏膜屏障功能并发挥抗氧化能力，可在替抗使用方面发挥积极作用。

关键词: 仔兔断奶腹泻, 加味枳术散, 肠黏膜屏障, 炎症反应, 抗氧化

Abstract:

The study aimed to investigate the effects of Modified Zhizhu Powder on the ileal intestinal mucosal barrier of weaned rabbits, with the goal of providing new ideas for the prevention and treatment of diarrhea diseases in weaned rabbits. Ninety 35-day-old New Zealand white rabbits, with similar body weights, were randomly divided into 5 groups, based on the principle of 50/50 male and female. Rabbits in control group were fed a basal diet, Enrofloxacin group, low-dose group, medium-dose group and high-dose group were fed the basal diet supplemented with 0.2% Enrofloxacin powder and 0.1%, 0.3% and 0.5% Modified Zhizhu Powder, respectively. The pre-test lasted for 7 days, and the formal test lasted for 14 days. The integrity of the intestinal barrier and its antioxidant capacity were measured after the experiment. The results showed that adding Modified Zhizhu Powder could significantly reduce the intestinal pathological tissue score (P < 0.05). Compared with the blank group, the other treatment groups all significantly increased the relative expression level of ZO2 (zonula occludens protein 2) mRNA in the ileum (P < 0.001). The high-dose group significantly increased the mRNA levels of Occludin, claudin, ZO1 (zonula occludens protein 1), ZO2, andJAM2 (junctional adhesion molecule 2) (P < 0.001). Modified Zhizhu Powder significantly increased the expression of MUC2 (Mucin 2) protein in the ileum (P < 0.05) and the high-dose group significantly reduced MPO (myeloperoxidase) activity (P < 0.01). Modified Zhizhu Powder high-doce group could significantly enhance the secretion of sIgA, reduce the levels of pro-inflammatory cytokinesIL-1β, IL-2, IL-6, andTNF-α, and increase the levels of anti-inflammatory cytokinesIL-10 andTGF-β. Modified Zhizhu Powder could also improve the activity of antioxidant enzymes (SOD, GSH, T-AOC, CAT). In conclusion, the Modified Zhizhu Powder could effectively improve the morphology and structure of the ileum of weaned rabbits, strengthen the tight connection between intestinal cells, maintain the permeability of intestinal mucosa, reduce the inflammatory response of the intestinal tract, enhance the function of the intestinal mucosal barrier, and play an antioxidant role, which can have a positive effect on the use of substitute antibodies.

Key words: weaning diarrhea of young rabbits, Modified Zhizhu Powder, intestinal mucosal barrier, inflammatory response, antioxidation

中图分类号:

S853.74

靳茹文, 王英杰, 蒋全兴, 刘天强, 邓阳, 罗杰, 赵玲, 叶刚, 施飞, 李英伦, 唐华侨. 加味枳术散对断奶仔兔肠黏膜屏障和抗氧化能力的影响[J]. 畜牧兽医学报, 2024, 55(12): 5825-5838.

JIN Ruwen, WANG Yingjie, JIANG Quanxing, LIU Tianqiang, DENG Yang, LUO Jie, ZHAO Ling, YE Gang, SHI Fei, LI Yinglun, TANG Huaqiao. Effects of Modified Zhizhu Powder on Intestinal Mucosal Barrier and Antioxidant Capacity of Weaned Rabbits[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5825-5838.

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原料 Ingredient	含量 Content	营养水平² Nutrient level	含量 Content
二级玉米 Two gard corn	12	消化能/(MJ·kg^-1) DE	12.65
豆粕 Soybean meal	5.5	总灰分 Total ash	8.63
膨化大豆 Expanded soybean	2.5	粗蛋白 Crude protein	15.48
玉米胚芽粕 Corn germ meal	12	粗脂肪 Crude fat	4.29
菜籽粕 Rapeseed meal	6	粗纤维 Crude fiber	13.76
苜蓿草颗粒 Alfalfa pellets	22.5	钙 Ca	1.02
统糠 Chaff	13	磷 P	0.51
小麦次粉 Wheat middlings	5	赖氨酸 Lysine	0.79
小麦麸皮 Wheat bran	17	蛋氨酸 Methionine	0.33
302肉兔核心料¹ 302 Meat rabbit core material	2	半胱氨酸 Cystine	0.21
石粉 Limestone	1.1
磷酸氢钙 Ca(HCO₃)₂	0.7
大豆油 Soybean oil	0.7
合计 Total	100

组别 Group	日粮组成 Diet composition	试验动物数量/只 Number of experimental animals
空白组 Control group	基础饲粮	18
恩诺沙星组 Enrofloxacin group	基础日粮+0.2%恩诺沙星粉剂	18
低剂量组 Low-dose group	基础日粮+0.1%加味枳术散粉剂	18
中剂量组 Medium-dose group	基础日粮+0.3%加味枳术散粉剂	18
高剂量组 High-dose group	基础日粮+0.5%加味枳术散粉剂	18

评分 Score	病变情况 Pathological condition
评分 Score	黏膜上皮 Mucosal epithelium	固有层 Proper layer
0	未见脱落坏死	绒毛结构清晰
1	无	毛细管充轻微血
2	上皮纹状缘萎缩消失	毛细管充血，炎细胞浸润
3	黏膜上皮细胞脱落坏死	脂肪细胞增生，炎细胞浸润，形成空泡
4	上皮细胞脱落坏死，较重	炎细胞浸润明显
5	绒毛结构破坏，上皮细胞大面积脱落坏死	脂肪空泡较多，炎细胞浸润明显

组分名称 Constituent	体积/μL Volume
cDNA	2
上游引物 Forwad primer	0.5
下游引物 Reverse primer	0.5
SYBR Green Master Mix	5
ddH₂O	2
合计 Total	10

引物名称 Primer name	登录号 Accession No.	引物序列(5′→3′) Primer sequence(5′→3′)
GAPDH	NM_001082253	F：TCGGAGTGAACGGATTTGGC R：GCCGTGGGTGGAATCATAC
ZO2	XM_017341711	F：GAGGAGAGACGCCAGCA R：GGAGGACGTGAACGGAGT
ZO1	XM_017348360	F：TTTCGTGTTGTGGATACCTT R：TGTATACTGCACACTGG
JAM2	XM_017346699	F：TGATAGATTTCAGTATACGCATC R：GGCACTAACTTCACAACGA
Claudin	XM_017347162	F：GTTGCCAGTTCCTCTTACCTT R：AGATACAGGCACAAAGTCCA
Occludin	XM_017344772	F：TGTTTATTGCCACCATTGTCT R：GTGAGCCATACATAGATCCAG
IL-1β	NM_001082201	F：CAAGAGGCACAACAGATCGC R：CGGCCTCAAAGAACAGGTCA
TNF-α	XM_008262537	F：CACTTCAGGGTGATCGGC R：TGCGGGTTTGCTACTACG
IL-2	NM_001163180	F：CATCATCTCAGCATCGAGTTCA R：TGAAGCCATTTTAGAGCCCCT
IL-6	NM_001082064	F：ACCTGCCTGCTGAGAATCAC R：TCGTCACTCCTGAACTTGGC
IL-10	NM_001082045	F：TCCCTCTTACCCTCTGGAGC R：AAGGGGTTGAAAATCGAGGTCA
TGF-β11	XM_008249704	F：CTGGAACGGGCTCAACATCT R：CAGGTCCTTGCGGAAGTCAA

加味枳术散对断奶仔兔肠黏膜屏障和抗氧化能力的影响

Effects of Modified Zhizhu Powder on Intestinal Mucosal Barrier and Antioxidant Capacity of Weaned Rabbits

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