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Identification and Analysis of Gastrodia elata miRNA and Preliminary Studies on Its Crosskingdom Regulation

Chunxin Xia  Shouliang Li  Xiaoyuan Xu  Tianlong Jiang  Huaixiang Zhou  Qing Sheng  
【摘要】:Gastrodia elata is one of precious Chinese herbal medicines,which has a variety of functions such as anticonvulsant effect,neuroprotection,anti-inflammatory and immune regulation.MicroRNAs(miRNAs) are a type of small,non-coding,19-24 nt biological RNA molecules.In this study,5718 known miRNAs were found,of which 52 were new miRNAs by Illumina high-throughput sequencing.The relative expression level of 10 selected candidate miRNAs was further validated by qRTPCR,and the results were consistent with the results of high throughput sequencing.A series of studies on Gas-mir01 and Gas-mir02 of Gastrodia elata with high expression and stability were carried out.The result showed that the relative levels of Gas-miR01 and Gas-miR02 significantly increased in each tissue of experimental mice that were orally administered,demonstrating that Gas-miR01 and Gas-miR02 can enter the mouse through the intestine.And then we found that both Gas-miR01 mimics and Gas-miR02 mimics could down-regulate the key protein A20 of NF-κB signaling pathway in 293 T cells.A20 is a NF-κB-dependent protein.When NF-κB is activated in a large amount,A20 can be phosphorylated by IKKβ and inhibit the activation of NF-κB,which plays a role in feedback regulation.This discovery provides a new path for the further development of Gastrodia elata anti-inflammatory drugs.It also provides a scientific basis for further research on the mechanism of miRNA action of Gastrodia elata.

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