Hock Alison, Miyake Hiromu, Li Bo, Lee Carol, Ermini Leonardo, Koike Yuhki, Chen Yong, Määttänen Pekka, Zani Augusto, Pierro Agostino
Division of General and Thoracic Surgery, Physiology and Experimental Medicine Program, The Hospital for Sick Children, Toronto, ON, Canada.
Physiology and Experimental Medicine Program, The Hospital for Sick Children, Toronto, ON, Canada.
J Pediatr Surg. 2017 May;52(5):755-759. doi: 10.1016/j.jpedsurg.2017.01.032. Epub 2017 Jan 29.
Breast milk administration prevents necrotizing enterocolitis (NEC). However, the mechanism remains unclear. Exosomes are cell-derived vesicles highly present in human milk and regulate intercellular signaling, inflammation, and immune response. We hypothesized that milk-derived exosomes beneficially affect intestinal epithelial cells.
Rat milk was collected, and exosomes were isolated using ExoQuick reagent and visualized by Nanoparticle Tracking Analysis. Protein was extracted from encapsulating exosomes, and concentration was measured. 2×10 intestinal epithelial cells (IEC-18) were treated for five hours with 0.5-μg/μl exosomes, an equal volume of exosome-free milk, or control solution (PBS). IEC-18 viability was measured using a colorimetric assay (MTT), and gene expression was analyzed by qRT-PCR. Data were compared using one-way ANOVA with Bonferroni post-test.
Rat milk was collected, and exosome isolation was confirmed. Compared to control, treatment with exosomes significantly increased IEC viability, proliferation, and stem cell activity (all p<0.05). However, administration of exosome-free milk had less significant effects.
Rat milk-derived exosomes promote IEC viability, enhance proliferation, and stimulate intestinal stem cell activity. These findings provide insight into the mechanism of action of breast milk in the intestines. Exosome administration is a promising prevention method for infants at risk of developing NEC when breastfeeding is not tolerated.
母乳喂养可预防坏死性小肠结肠炎(NEC)。然而,其机制尚不清楚。外泌体是细胞来源的囊泡,在人乳中大量存在,可调节细胞间信号传导、炎症和免疫反应。我们推测母乳来源的外泌体对肠道上皮细胞有有益影响。
收集大鼠乳汁,使用ExoQuick试剂分离外泌体,并通过纳米颗粒跟踪分析进行可视化。从包裹外泌体中提取蛋白质并测量浓度。将2×10个肠道上皮细胞(IEC-18)用0.5μg/μl外泌体、等体积的无外泌体乳汁或对照溶液(PBS)处理5小时。使用比色法(MTT)测量IEC-18活力,并通过qRT-PCR分析基因表达。数据使用单因素方差分析和Bonferroni事后检验进行比较。
收集大鼠乳汁并确认外泌体分离。与对照组相比,用外泌体处理显著提高了IEC活力、增殖和干细胞活性(所有p<0.05)。然而,给予无外泌体乳汁的效果不太显著。
大鼠乳汁来源的外泌体促进IEC活力,增强增殖,并刺激肠道干细胞活性。这些发现为母乳在肠道中的作用机制提供了见解。当无法耐受母乳喂养时,外泌体给药是预防有发生NEC风险婴儿的一种有前景的方法。