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Toll样受体4(TLR 4)单核苷酸多态性对奶牛繁殖性能和免疫功能的影响

Effect of Single Nucleotide Polymorphisms of Toll-Like Receptor 4 (TLR 4) on Reproductive Performance and Immune Function in Dairy Cows.

作者信息

Shimizu Takashi, Kawasaki Yurie, Aoki Yuka, Magata Fumie, Kawashima Chiho, Miyamoto Akio

机构信息

Graduate School of Animal and Food Hygiene, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, 0808555, Japan.

Field Center of Animal Science and Agriculture, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, 0808555, Japan.

出版信息

Biochem Genet. 2017 Jun;55(3):212-222. doi: 10.1007/s10528-017-9790-0. Epub 2017 Jan 16.

Abstract

In dairy cows, inflammatory diseases caused by infection with pathogenic bacteria post calving affect ovarian functions. This study examined the relationship between single-nucleotide polymorphisms (SNPs) of Toll-like receptor 4 (TLR4), reproductive performances [the number of artificial insemination (AI) application and days open], and immune cell functions (apoptosis and migration). Two hundred Holstein cows from the Obihiro University farm were included. The SNPs of TLR4 were genotyped by PCR-restriction fragment length polymorphism (PCR-RFLP) method. Polymorphonuclear leukocytes (PMNs) and peripheral blood mononuclear cells (PBMCs) were isolated from whole blood. The number of AI application in the animals with T/C genotype in the TLR4 exon3 was lower than that in animals with C/C genotype (1.6 ± 0.2 and 2.2 ± 0.2, respectively). Among the animals with TLR4 exon3 polymorphisms, the days open was shorter for the T/C cows than that for C/C cows (100.7 ± 6.9 days and 136.6 ± 9.0 days, respectively). The SNPs in the TLR4 intron did not affect the number of AI and days open. The apoptosis percentage of PMNs treated with lipopolysaccharide (LPS; 0.001 and 1 μg/ml) tended to be lower in the T/C genotype compared to that in the C/C genotype. The transmigration rates of PMNs, and IL-1β production in PBMCs were tended to be higher for the animals with the T/C genotype compared to those for animals with the C/C genotype. Taken together, these results suggest that TLR4 polymorphisms offer a meaningful tool to judge the reproductive potential and immune activity in individual cows.

摘要

在奶牛中,产后感染病原菌引起的炎症性疾病会影响卵巢功能。本研究检测了Toll样受体4(TLR4)的单核苷酸多态性(SNP)、繁殖性能[人工授精(AI)次数和空怀天数]以及免疫细胞功能(凋亡和迁移)之间的关系。研究纳入了来自小牧大学农场的200头荷斯坦奶牛。采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法对TLR4的SNP进行基因分型。从全血中分离出多形核白细胞(PMN)和外周血单核细胞(PBMC)。TLR4外显子3中T/C基因型动物的AI次数低于C/C基因型动物(分别为1.6±0.2次和2.2±0.2次)。在具有TLR4外显子3多态性的动物中,T/C基因型奶牛的空怀天数比C/C基因型奶牛短(分别为100.7±6.9天和136.6±9.0天)。TLR4内含子中的SNP不影响AI次数和空怀天数。与C/C基因型相比,用脂多糖(LPS;0.001和1μg/ml)处理的PMN的凋亡百分比在T/C基因型中趋于更低。与C/C基因型动物相比,T/C基因型动物的PMN迁移率和PBMC中IL-1β的产生趋于更高。综上所述,这些结果表明TLR4多态性为判断个体奶牛的繁殖潜力和免疫活性提供了一个有意义的工具。

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