乳房炎奶牛血浆源外泌体对LPS诱导的MAC-T细胞炎症的影响

doi:10.16431/j.cnki.1671-7236.2024.11.044

Abstract

Abstract: 【Objective】 Exosomes are key factors in cellular communication and establishment of tissue microenvironment,and they have a regulatory role in inflammatory processes.This study was aimed to investigate the role of plasma-derived exosomes from dairy cows in the regulation of lipopolysaccharide (LPS)-induced inflammation in mammary epithelial cells (MAC-T) of dairy cows. 【Method】 In this study,venous blood was collected from healthy and mastitis-affected dairy cows,and plasma was obtained after centrifugation,and plasma-derived exosomes were isolated by ultracentrifugation.The expression of pro-inflammatory factors after incubation of plasma-derived exosomes from dairy cows was detected by Real-time quantitative PCR.Western blotting was used to verify the expression of NF-κB pathway proteins TLR4,MyD88 and TNF-α after incubation of plasma-derived exosomes from dairy cows.The effects of plasma-derived exosomes from dairy cows on the proliferation as well as apoptosis of MAC-T cells were verified by CCK8 and Hoechst staining. 【Result】 Real-time quantitative PCR results showed that plasma-derived exosomes from dairy cows with mastitis significantly down-regulated the expression of pro-inflammatory factors IL-6,IL-23,IL-1α,NFRK,MyD88,TNF-α and IL-1β genes mRNA compared with LPS group (P＜0.05). Plasma-derived exosomes from healthy cows significantly inhibited mRNA expression of the IL-1β gene (P＜0.05).Western blotting results showed that the plasma-derived exosomes from dairy cows with mastitis significantly decreased the expression of the NF-κB pathway proteins TLR4,MyD88 and TNF-α (P＜0.05). NF-κB pathway protein expression in plasma-derived exosomes from healthy cows was not significantly different from that of the LPS group (P＞0.05). CCK8 and Hoechst staining results showed that plasma-derived exosomes from dairy cows with mastitis could promote cell proliferation and alleviate apoptosis compared with LPS group. Plasma-derived exosomes from healthy cows restored some cell viability. 【Conclusion】 The results indicated that plasma-derived exosomes from dairy cows with mastitis could alleviate LPS-induced inflammation of MAC-T cells through NF-κB pathway.This study would provide a theoretical basis for the anti-inflammatory role played by plasma-derived exosomes in dairy cow mastitis,and at the same time laid the foundation for the clinical application of plasma-derived exosomes.

Key words: plasma-derived exosomes; NF-κB pathway; anti-inflammatory; dairy cow mastitis

CLC Number:

S852.3

LI Yang, WANG Zhaoyuan, HE Xingli, LIU Peng, WANG Zi, LI Xiaolin, SONG Yangyang, SHEN Binglei. Effect of Plasma-derived Exosomes on LPS-induced MAC-T Cell Inflammation in Dairy Cows with Mastitis[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(11): 5097-5107.

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Effect of Plasma-derived Exosomes on LPS-induced MAC-T Cell Inflammation in Dairy Cows with Mastitis

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