Exosomes Derived from Bovine Mammary Epithelial Cells Treated with Transforming Growth Factor-beta 1 Inhibit the Proliferation of Bovine Macrophages
Exosomes Derived from Bovine Mammary Epithelial Cells Treated with Transforming Growth Factor-beta 1 Inhibit the Proliferation of Bovine Macrophages
复制标题
用转化生长因子-β1处理的牛乳腺上皮细胞衍生的外泌体抑制牛巨噬细胞的增殖
DOI：
10.1089/jir.2019.0032
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发表时间：
2019
影响因子：
2.3
通讯作者：
Lei Liancheng
中科院分区：
文献类型：
--
作者：
Huang Tinghao;Zhou Changhai;Che Yanyi;Zhang Meina;Ren Wenbo;Lei Liancheng
转化生长因子（TGF）-β1是一种多功能细胞因子，在调节免疫细胞增殖中发挥重要作用。我们推测TGF-β1的高表达可能会影响奶牛的免疫力。在本研究中，通过超速离心分离来自未处理的牛乳腺上皮细胞系（MAC-T）的未经处理的外泌体（un-exo）和来自经TGF-β1处理的MAC-T细胞的经TGF-β1处理的外泌体（t-exo），并通过电子显微镜和蛋白质印迹进行鉴定。然后，使用un-exo和t-exo处理牛巨噬细胞系(BOSMAC)，并通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴化物测定和流式细胞术检测BOSMAC细胞的增殖能力。通过q-PCR和Western blotting分析p38的表达和磷酸化水平。结果表明，两种外泌体类型均表现出外泌体的基本特征。在用t-exo处理的BOSMAC细胞中，观察到细胞增殖的显着抑制，并且细胞周期进程受到抑制，而un-exo组和对照组之间没有发现差异。仅用 t-exo 处理会增加 p38 的表达和磷酸化，并且添加 p38 抑制剂 SB203580 消除了 t-exo 对 BOSMAC 细胞增殖的抑制。我们的结果表明，t-exo 通过刺激 p38MAPK 抑制牛巨噬细胞的增殖，并可能干扰奶牛的免疫力。这一发现可能为提高奶牛免疫力和预防乳腺相关疾病提供新策略。
Transforming growth factor (TGF)-β1 is a multifunctional cytokine that plays an important role in regulating immune cell proliferation. We speculate that high expression ofTGF-β1may affect the immunity of dairy cows. In this study, untreated exosomes (un-exo) derived from an untreated bovine mammary epithelial cell line (MAC-T) andTGF-β1-treated exosomes (t-exo) derived fromTGF-β1-treated MAC-T cells were isolated by ultracentrifugation and identified by electron microscopy and Western blotting. Then, un-exo and t-exo were used to treat a bovine macrophage cell line (BOSMAC), and the proliferative ability of BOSMAC cells was detected by an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and flow cytometry. The expression and phosphorylation levels of p38 were analyzed by q-PCR and Western blotting. The results showed that both exosome types exhibited the basic characteristics of exosomes. In BOSMAC cells treated with t-exo, significant inhibition of cell proliferation was observed, and the cell cycle progression was inhibited, while no difference was found between the un-exo and control groups. Only treatment with t-exo increased the expression and phosphorylation of p38, and the addition of the p38 inhibitor SB203580 abrogated the inhibition of BOSMAC cell proliferation by t-exo. Our results demonstrated that t-exo inhibited the proliferation of bovine macrophages by stimulating p38MAPKand might interfere with immunity in dairy cattle. This finding may provide a new strategy for improving immunity and preventing breast-related diseases in dairy cows.