Protective effects of bovine milk exosomes against oxidative stress in IEC-6 cells
Protective effects of bovine milk exosomes against oxidative stress in IEC-6 cells
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DOI：
10.1007/s00394-020-02242-z
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发表时间：
2020-04-23
影响因子：
5
通讯作者：
Li, Ping
中科院分区：
文献类型：
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作者：
Wang, Lanfang;Shi, Zhexi;Li, Ping
目的牛乳外泌体富含microRNA（miRNAs）和蛋白质，调节免疫应答和生长。在本研究中，我们旨在评估牛乳外泌体对肠隐窝上皮细胞（IEC-6）氧化应激的保护作用。方法对牛乳外泌体进行分离和鉴定。为了评估外泌体的保护作用，用外泌体预处理IEC-6细胞，然后用H2 O2预处理。测定细胞活力和超氧化物歧化酶（SOD）、丙二醛（MDA）、谷胱甘肽过氧化物酶（GPX）、活性氧化物质（ROS）和乳酸脱氢酶（LDH）水平。还测定了核因子红细胞2相关因子2（Nrf 2）和血红素加氧酶1（Ho 1）基因以及miR-146 a、miR-155和HO-1蛋白的表达水平。结果分离的牛乳外泌体表达CD 63和CD 9。外泌体近似为圆形，直径约为67.23 nm。用牛乳外泌体预处理IEC-6细胞增强了细胞活力;增加了SOD和GPX活性;并在H2 O2攻击后降低了LDH、ROS和MDA水平。进一步的分析显示，外泌体预处理增加了细胞内miR-146 a和miR-155水平。Exosome预处理抑制H2 O2诱导的Nrf 2和Ho 1基因表达的升高，但促进HO-1蛋白的表达。结论牛乳外泌体对IEC-6细胞氧化应激具有保护作用。
Purpose Bovine milk exosomes, which are enriched with microRNAs (miRNAs) and proteins, regulate immune response and growth. In the present study, we aimed to assess the protective effects of bovine milk exosomes against oxidative stress of intestinal crypt epithelial cells (IEC-6). Methods Bovine milk exosomes were isolated and characterized. To assess the protective effects of exosomes, IEC-6 cells were pretreated with exosomes, followed by H2O2. Cell viability and levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPX), reactive oxidative species (ROS), and lactate dehydrogenase (LDH) were measured. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (Ho1) genes, and miR-146a, miR-155, and the HO-1 protein were also determined. Results The isolated bovine milk exosome were positive for CD63 and CD9 expression. The exosomes were approximately circular and had a diameter of about 67.23 nm. Pretreatment of IEC-6 cells with bovine milk exosomes enhanced cell viability; increased SOD and GPX activities; and reduced LDH, ROS, and MDA levels after H2O2 challenge. Further analysis showed that exosome pretreatment increased intracellular miR-146a and miR-155 levels. Exosome pretreatment inhibited the elevation of Nrf2 and Ho1 gene expression induced by H2O2, but promoted HO-1 protein expression. Conclusion The results indicated that bovine milk exosomes exerted protective effects against oxidative stress in IEC-6 cells.