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基于抑制性消减杂交技术筛选益生菌枯草芽孢杆菌 PAS38 对肉鸡原代淋巴器官免疫相关差异表达基因的研究。

Screening of immune-related differentially expressed genes from primary lymphatic organs of broilers fed with probiotic bacillus cereus PAS38 based on suppression subtractive hybridization.

机构信息

College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan Province, China.

Branch of Animal Husbandry and Veterinary Medicine, Chengdu Vocational College of Agricultural Science and Technology, Chengdu, Sichuan Province, China.

出版信息

PLoS One. 2020 Jul 1;15(7):e0235476. doi: 10.1371/journal.pone.0235476. eCollection 2020.

Abstract

To explore the molecular mechanism of the effect of Bacillus cereus PAS38 on the immunity of broilers, sixty 7-day-old broilers were divided into two groups with three replicates. The control group was fed with basal diet, and the treatment group was fed with basal diet containing Bacillus cereus PAS38 1×106 CFU/g. Thymus and bursa of fabricius were taken from two groups of broilers at the age of 42 days, total RNA was extracted, differential gene library was constructed by SSH technology, and immune-related differential genes were screened. Then, we used siRNA to interfere with the expression of some differential genes in the original generation lymphocytes of broiler blood to detect the change of cytokines mRNA expression level. A total of 42 immune-related differentially expressed genes were screened, including 22 up-regulated genes and 20 down-regulated genes. When 7 differentially up-regulated genes associated with enhanced immune function were interfered with in lymphocytes, some immune-promoting cytokines were down-regulated. These results showed that Bacillus cereus PAS38 might up-regulate the expression of JCHAIN, PRDX1, CD3E, CDK6 and other genes in immune organs of broilers, thereby affecting the development of immune organs, the expression of various cytokines and the transduction of immune signals, improving the immune capacity of broilers.

摘要

为了探索蜡样芽胞杆菌 PAS38 对肉鸡免疫的作用机制,将 60 只 7 日龄肉鸡分为两组,每组 3 个重复。对照组饲喂基础日粮,试验组饲喂含 1×106 CFU/g 蜡样芽胞杆菌 PAS38 的基础日粮。在 42 日龄时从两组肉鸡中取出胸腺和法氏囊,提取总 RNA,采用 SSH 技术构建差异基因文库,筛选与免疫相关的差异基因。然后,我们使用 siRNA 干扰肉鸡原代淋巴细胞中某些差异基因的表达,检测细胞因子 mRNA 表达水平的变化。共筛选到 42 个与免疫相关的差异表达基因,其中上调基因 22 个,下调基因 20 个。当干扰淋巴细胞中与增强免疫功能相关的 7 个差异上调基因时,一些免疫促进细胞因子的表达下调。这些结果表明,蜡样芽胞杆菌 PAS38 可能通过上调肉鸡免疫器官中 JCHAIN、PRDX1、CD3E、CDK6 等基因的表达,从而影响免疫器官的发育、各种细胞因子的表达和免疫信号的转导,提高肉鸡的免疫能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a1d/7329121/19afbba6dc7a/pone.0235476.g001.jpg

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