目的 为更好了解药用黄芪微小RNA(miRNA)被机体摄入和吸收的情况,试图建立黄芪miRNA谱,为今后黄芪饮片药理学研究奠定基础。方法 以经干燥、高温和微波处理的黄芪煎煮液为样本,采用高通量深度测序方法以获取未知的miRNA序列。结果 经测序得到药用黄芪9 931 049个小RNA读数。经与miRBase 21数据库所有植物种类潜在发卡型结构比对,分析和构建了黄芪饮片煎煮液miRNA谱,并确定了1个保守的miRNA和9个新发现的miRNA。经进一步生物学信息分析,发现12种I类小干扰性RNA(small interfering RNAs,siRNAs)序列,其5'端对热力学不稳定,3'端种子区域对热力学高度稳定。结论 首次揭示了经热力学处理后黄芪饮片的miRNA谱,I类siRNAs是潜在的可利用的植物药的生物化学分子,有助于今后中药在表观遗传学方面的深入研究。

Objective To establish a small RNA library of Astragalus membranaceus, in order to understand the intake and absorption of miRNAs in human body, and to lay the foundation for pharmacological research of Astragali Radix decoction pieces (ARP). Methods The decoction of A. membranaceus after desiccation, high temperature and microwave treatment was used as sample, and using high-throughput sequencing technology, the unknown miRNA sequence was obtained. Results Totally 9 931 049 small RNA sequences were identified from ARP by the high-throughput sequencing technology. After compared with all hairpin-forming precursors in miRBase 21 database, the miRNA library of ARP decoction was established, and one conserved miRNA and 9 novel miRNA sequences were confirmed. Twelve class I small interfering RNAs (siRNAs) were obtained by further bioinformatic analysis and we found that the 5' terminal was unstable while the 3' terminal was stable thermodynamically. Conclusion The miRNAs expression profiles of ARP is first revealed by thermodynamic treatment. Our study suggests that class I siRNA can be used as a potential biochemistry components of phytomedicine, which might contribute to the epigenetic studies on Chinese materia medica.

上海市中医药发展办公室中西医结合重点专病基金资助项目(zxbz2012-003)