Research Center for Translational Medicine at Shanghai East Hospital, School of Life Science and Technology , Tongji University , 1239 Siping Road , Shanghai 200092 , People's Republic of China.
J Agric Food Chem. 2018 Mar 21;66(11):2749-2757. doi: 10.1021/acs.jafc.7b05614. Epub 2018 Mar 2.
In this study, the presence of microRNAs in coconut water was identified by real-time polymerase chain reaction (PCR) based on the results of high-throughput small RNA sequencing. In addition, the differences in microRNA content between immature and mature coconut water were compared. A total of 47 known microRNAs belonging to 25 families and 14 new microRNAs were identified in coconut endosperm. Through analysis using a target gene prediction software, potential microRNA target genes were identified in the human genome. Real-time PCR showed that the level of most microRNAs was higher in mature coconut water than in immature coconut water. Then, exosome-like nanoparticles were isolated from coconut water. After ultracentrifugation, some particle structures were seen in coconut water samples using 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate fluorescence staining. Subsequent scanning electron microscopy observation and dynamic light scattering analysis also revealed some exosome-like nanoparticles in coconut water, and the mean diameters of the particles detected by the two methods were 13.16 and 59.72 nm, respectively. In conclusion, there are extracellular microRNAs in coconut water, and their levels are higher in mature coconut water than in immature coconut water. Some exosome-like nanoparticles were isolated from coconut water, and the diameter of these particles was smaller than that of animal-derived exosomes.
在这项研究中，通过高通量小 RNA 测序的结果，使用实时聚合酶链反应（PCR）鉴定了椰水中 microRNA 的存在。此外，还比较了未成熟和成熟椰水中 microRNA 含量的差异。在椰子胚乳中鉴定出 47 种已知的 microRNA，属于 25 个家族，还有 14 种新的 microRNA。通过使用靶基因预测软件进行分析，在人类基因组中鉴定出潜在的 microRNA 靶基因。实时 PCR 显示，成熟椰水中大多数 microRNA 的水平高于未成熟椰水中的水平。然后，从椰水中分离出外泌体样纳米颗粒。经过超速离心后，使用 1,1'-二辛基-3,3,3',3'-四甲基吲哚碳菁高氯酸盐荧光染色，在椰水样品中观察到一些颗粒结构。随后的扫描电子显微镜观察和动态光散射分析也揭示了椰水中的一些外泌体样纳米颗粒，两种方法检测到的颗粒的平均直径分别为 13.16nm 和 59.72nm。总之，椰水中存在细胞外 microRNA，其在成熟椰水中的水平高于未成熟椰水中。从椰水中分离出一些外泌体样纳米颗粒，这些颗粒的直径小于动物来源的外泌体。
