Regulation of Glycine on Mammary Inflammatory Response in Dairy Goats and Its Mechanism
DU Ruiping1, YUN Fuyu2, ZHANG Xingfu1, SONG Liwen1, GAO Min1
1. Animal Nutrition Institute of Agriculture and Animal Husbandry Academy of Inner Mongolia, Huhhot 010031, China;
2. Agriculture and Animal Husbandry Academy of Ulanchabu, Jining 012000, China
Abstract：This experiment was conducted to investigate the regulation of glycine on mammary inflammatory response in dairy goats in vivo and its signaling pathway changes. Fifteen healthy lactating Saanen dairy goats with similar body weight, lactation period and milk yield were selected and the goats were randomly divided into 5 groups with 3 goats in each group. Dairy goats in lipopolysaccharide (LPS) model group were infused 1 mL LPS solution (4 μg/mL) via each udder; dairy goats in glycine intervention groups were infused three doses[1, 5 and 25 g/(goat·d)] of glycine separately which lasting 5 days, and 1 mL LPS solution (4 μg/mL) via each udder were infused on the sixth day; dairy goats in control group were infused without glycine and LPS, but were infused 1 mL normal saline. Dairy goats in each group were slaughtered and sampling 24 h after LPS or normal saline infusion. Real time quantitative PCR method was used to assay the gene expression levels of interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), Toll like receptor (TLR)2, TLR4, TLR9, myeloid differentiation factor 88 (MyD88) and TIR-domain-containing adapter-inducing interferon-β (TRIF) in mammary tissue, the Western blotting method was used to detect the protein expression level of nuclear transcription factor-κB (NF-κB) p65 in mammary tissue, and the mammary tissue section and hematoxylin-eosin (HE) staining was used to observe the changes of histopathology. The results showed that the goats' body temperature of the glycine intervention groups decreased below 39.5℃ at 24 h after LPS infusion. Compared with LPS model group, the inflammatory cell invasion and necrosis of mammary gland cells were obviously reduced in the 3 glycine intervention groups; the gene expression levels of IL-1β, IL-8 and COX-2 were significantly decreased in the low, middle and high dose glycine intervention groups (P<0.05), and the gene expression levels of IL-6 and TNF-α were significantly decreased in middle dose glycine intervention group (P<0.05); the gene expression level of TLR4 was significantly decreased in the low, middle and high dose glycine intervention groups (P<0.05), and the gene expression levels of TLR2 and TLR9 were significantly decreased in middle dose glycine intervention group (P<0.05); the gene expression level of MyD88 was significantly decreased in the low, middle and high dose glycine intervention groups (P<0.05), and the gene expression level of TRIF was significantly decreased in low and high dose glycine intervention groups (P<0.05); the protein expression level of NF-κB p65 was significantly decreased in the low, middle and high dose glycine intervention groups (P<0.05). The results demonstrate that glycine reduces the secretion of inflammatory factors IL-1β, IL-6, IL-8, TNF-α and COX-2 by down-regulating the gene expression of TLR4 and its downstream key signaling molecules MyD88 and TRIF, and inhibiting the phosphorylation of nuclear transcription factor NF-κB p65. By the results, we preliminarily illuminate that glycine inhibits the activation or activity of inflammatory cells by inhibiting TLR-NF-κB signaling pathway, and prevents the potential inflammatory mediators releasing, such as toxic cytokines and prostaglandins, thus hinders the realization of inflammatory response.
杜瑞平, 云伏雨, 张兴夫, 宋利文, 高民. 甘氨酸对奶山羊乳腺炎性应答的调节及其作用机制研究[J]. 动物营养学报, 2019, 31(7): 3110-3122.
DU Ruiping, YUN Fuyu, ZHANG Xingfu, SONG Liwen, GAO Min. Regulation of Glycine on Mammary Inflammatory Response in Dairy Goats and Its Mechanism. Chinese Journal of Animal Nutrition, 2019, 31(7): 3110-3122.
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