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基于盲肠内容物肠道菌群及短链脂肪酸检测与关联分析的焦糊层多糖对蛋鸡的保肝作用机制研究

Study on the Hepatoprotective Effect Mechanism of Polysaccharides from Charred on the Layer Chickens Based on the Detection of the Intestinal Floras and Short-Chain Fatty Acids of Cecal Contents and Association Analysis.

作者信息

Wu Fanlin, Ji Peng, Hu Yonghao, Li Chenchen, He Jian

机构信息

College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China.

出版信息

Vet Sci. 2023 Mar 15;10(3):224. doi: 10.3390/vetsci10030224.

DOI:10.3390/vetsci10030224
PMID:36977263
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10058873/
Abstract

To analyze the intervention mechanism of polysaccharides from charred (CASP) on the liver injury caused by Ceftiofur sodium (CS) and lipopolysaccharide (LPS) from the perspective of the intestine. Ninety-four one-day-old laying chickens underwent free feeding and drinking water for three days. Then, fourteen laying chickens were randomly selected as the control group, and sixteen laying chickens were selected as the model group. Sixteen laying chickens in the rest were randomly selected as the intervention group of CASP. Chickens in the intervention group were given CASP by the oral administration (0.25 g/kg/d) for 10 days, the control and model groups were given the same amount of physiological saline. During the 8th and 10th days, laying chickens in the model and CASP intervention group were subcutaneously injected with CS at the neck. In contrast, those in the control group were subcutaneously injected with the same amount of normal saline simultaneously. Except for the control group, the layer chickens in the model and CASP intervention groups were injected with LPS after CS injection on the 10th day of the experiment. In contrast, those in the control group were injected with the same amount of normal saline at the same time. 48 h after the experiment, the liver samples of each group were collected, and the liver injury was analyzed by hematoxylin-eosin (HE) staining and transmission electron microscopy. And the cecum contents of six-layer chickens in each group were collected, and the intervention mechanism of CASP on the liver injury from the perspective of the intestine was analyzed by the 16S rDNA amplicon sequencing technology and the short-chain fatty acids (SCFAs) detection of cecal contents based on Gas Chromatography-Mass Spectrometry (GC-MS), and their association analysis was carried out. The results showed that the structure of chicken liver in the normal control group was normal, while that in the model group was damaged. The structure of chicken liver in the CASP intervention group was similar to the normal control group. The intestinal floras in the model group were maladjusted compared to the normal control group. After the intervention of CASP, the diversity, and richness of chicken intestinal floras changed significantly. It was speculated that the intervention mechanism of CASP on the chicken liver injury might be related to the abundance and proportion of Bacteroidetes and Firmicutes. Compared with the model group, the indexes of ace, chao1, observed species, and PD whole tree of chicken cecum floras in the intervention group of CASP were significantly increased ( < 0.05). The contents of acetic acid, butyric acid, and total SCFAs in the intervention group of CASP were significantly lower than those in the model group ( < 0.05), and the contents of propionic acid and valeric acid in the intervention group of CASP were significantly lower than those in the model group ( < 0.05) and normal control group ( < 0.05). The correlation analysis showed that the changes in the intestinal floras were correlated with the changes in SCFAs in the cecum. It is confirmed that the liver-protecting effect of CASP is indeed related to the changes in the intestinal floras and SCFAs content in the cecum, which provides a basis for screening liver-protecting alternative antibiotics products for poultry.

摘要

从肠道角度分析焦糊多糖(CASP)对头孢噻呋钠(CS)和脂多糖(LPS)所致肝脏损伤的干预机制。94只1日龄蛋鸡自由采食和饮水3天。然后,随机选取14只蛋鸡作为对照组,16只蛋鸡作为模型组。其余蛋鸡中随机选取16只作为CASP干预组。干预组蛋鸡按0.25 g/kg/d口服给予CASP,连续10天,对照组和模型组给予等量生理盐水。在第8天和第10天,模型组和CASP干预组蛋鸡颈部皮下注射CS。相比之下,对照组同时皮下注射等量生理盐水。除对照组外,模型组和CASP干预组蛋鸡在实验第10天CS注射后注射LPS。相比之下,对照组同时注射等量生理盐水。实验48 h后,采集各组肝脏样本,通过苏木精-伊红(HE)染色和透射电子显微镜分析肝脏损伤情况。并采集各组6只蛋鸡的盲肠内容物,采用16S rDNA扩增子测序技术和基于气相色谱-质谱联用(GC-MS)的盲肠内容物短链脂肪酸(SCFAs)检测,从肠道角度分析CASP对肝脏损伤的干预机制,并进行相关性分析。结果显示,正常对照组蛋鸡肝脏结构正常,模型组受损。CASP干预组蛋鸡肝脏结构与正常对照组相似。与正常对照组相比,模型组肠道菌群失调。CASP干预后,蛋鸡肠道菌群的多样性和丰富度发生显著变化。推测CASP对蛋鸡肝脏损伤的干预机制可能与拟杆菌门和厚壁菌门的丰度和比例有关。与模型组相比,CASP干预组蛋鸡盲肠菌群的ace、chao1、观测物种数和PD全树指数显著升高(P<0.05)。CASP干预组乙酸、丁酸和总SCFAs含量显著低于模型组(P<0.05),CASP干预组丙酸和戊酸含量显著低于模型组(P<0.05)和正常对照组(P<0.05)。相关性分析表明,肠道菌群变化与盲肠SCFAs变化相关。证实CASP的保肝作用确实与盲肠肠道菌群和SCFAs含量变化有关,为筛选家禽保肝替代抗生素产品提供了依据。

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