| 引用本文： |
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徐小艳,牟 姝,史哲溪,等.牛奶消化前后外泌体的变化及其对细胞增殖的影响[J].同济大学学报（医学版）,2018,39(5):39-43. [点击复制]
- XU Xiao-yan,MU Shu,SHI Zhe-xi,et al.Stability of milk exosome after digestion in vitro and its effect on cell proliferation[J].同济大学学报（医学版）,2018,39(5):39-43. [点击复制]
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| 摘要: |
| 目的 探究超速离心法获取牛奶外泌体的最适离心力，探讨牛奶外泌体在体外消化模型中的稳定性及消化后的牛奶外泌体对大鼠肠隐窝上皮细胞IEC-6增殖的影响。方法 用体外消化模型消化牛奶，不同离心力超速离心分离牛奶外泌体，用Western印迹法、动态光散射、扫描电镜对消化前后获取的外泌体进行表征，CCK8实验分析外泌体对细胞增殖的影响。结果 牛奶消化后外泌体蛋白浓度明显降低，外泌体标记蛋白CD63和CD9在消化前后的分离产物中均为阳性。消化后的外泌体粒径大于未消化组，且粒径随离心力的升高而降低。扫描电镜下观察，消化前后的牛奶外泌体形态差别不大。用50μg/mL外泌体培养IEC-6细胞，结果显示牛奶消化前后的外泌体均对IEC-6细胞的增殖有促进作用，消化后离心(130000×g)获得的外泌体对细胞增殖的促进作用较为明显。结论 130000×g是获取牛奶外泌体的最适离心力。牛奶消化前后的外泌体在标记蛋白、形态上相似，表明牛奶中的外泌体能耐受体外消化模型的消化，牛奶消化后的外泌体仍能促进IEC-6细胞的增殖。 |
| 关键词: 外泌体 体外消化模型 超速离心 细胞增殖 |
| DOI：10.16118/j.1008-0392.2018.05.008 |
| 投稿时间：2018-04-04 |
| 基金项目:国家自然科学基金（21476176）；国家高技术研究发展（863）计划（2015AA021002） |
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| Stability of milk exosome after digestion in vitro and its effect on cell proliferation |
| XU Xiao-yan,MU Shu,SHI Zhe-xi,WANG Lan-fang,LI Ping |
| (Tongji University School of Medicine, Shanghai 200092, China;Tongji University School of Life Sciences and Technology, Shanghai 200092, China) |
| Abstract: |
| Objective To explore the stability of bovine milk exosomes after digestion in vitro and their effects on cell proliferation. Methods Bovine milk exosomes were isolated by differential centrifugal speed and the characteristics of the exosomes were analyzed by Western blotting, DLS and SEM. The effects of milk exosomes on cell proliferation was determined by CCK8 assay. Results Protein concentration of exosomes in bovine milk was reduced after in vitro digestion, while the protein markers CD63 and CD9 were detected in exosomes both from undigested and digested milk. The size of exosomes derived from digested milk was larger than that from undigested milk; and the size of the exosomes in two groups was decreased with the increase of the relative centrifugal force. The exosomes extracted from the digested milk and the undigested milk were able to promote the proliferation of IEC-6 cells.
Conclusion To obtain bovine milk exosomes, the optimal centrifugal force is(130000×g RCF). The exosomes derived from digested milk are similar with exosomes derived from undigested milk in protein markers, size, shape, as well as the ability to promote cell proliferation, which suggests that milk exosomes are stable under in vitro digestion model. |
| Key words: milk exosomes in vitro digestion model ultracentrifugation cell proliferation |
