Research On The Effect Of Gastrodia Data Blume Specific MicroRNA On NF-κB Signaling Pathway

Gastrodia elata Blume is a perennial herb of the genus Orchidaceae.As a traditional Chinese medicine,Gastrodia elata Blume(G.elata)has a long history,which is widely used to treat diseases such as convulsions,dizziness,headache,numbness of limbs,rheumatism arthralgia and so on.Micro RNA(mi RNA)is a class of noncoding small RNA that regulate posttranscriptional silencing of target genes,and its mechanisms involve a variety of key biological and cellular processes,including metabolism,signal transduction pathways,etc.Studies have shown that plant mi RNAs have access to animals and play an important biological role.NF-κB signaling pathway is a key regulatory pathway for immunity,development and cancer,and is widely involved in cellular responses to various stimuli,especially in inflammation.So far,research at home and abroad on G.elata mainly focuses on the medicinal components and functions of G.elata,but there are few studies on micro RNAs in G.elata.On the basis of the previous research,the cross-kingdom regulation of G.elata mi RNA and its effect on NF-κB signaling pathway were deeply studied,which provide evidence for the mechanism on cross-kingdom regulation of mi RNA,and also lays a foundation for the study of anti-inflammatory effects of G.elata.In order to detect whether G.elata-unique mi RNAs exist in various tissues of mice,the contents of Gas-mi R01 and Gas-mi R02 were detected by extracting RNA from animal tissues and fluorescence PCR.The results showed that the G.elata-unique Gasmi R01 and Gas-mi R02 were detected in all tissues of mice,indicating that Gas-mi R01 and Gas-mi R02 could be into the mice by oral administration and the contents in different tissues were significantly different at any time.Then,we conducted in vitro cell experiments to verify whether Gas-mi R01 can regulate the NF-κB signaling pathway and attenuate inflammation.In our research,the m RNA level of inflammatory factors,and the level of related proteins on NF-κB signaling pathway in BV2 cells after LPS-induction are detected by q RT-PCR、Western Blotting and ELISA,respectively.It was found that Gas-miR01 could downregulate the expression of inflammatory factors in BV2 cells and inhibit the activation of NF-κB signaling pathway.The above results indicate that Gas-mi R01 can inhibit LPS-induced neuroinflammation by regulating NF-κB signaling pathway.To further verify the regulation function of Gas-mi R01,we constructed an animal neuroinflammation model.We used q RT-PCR,ELISA and Western Blotting to detect the levels of inflammatory factors and related proteins on the NF-κB pathway in the brain of mice.The results showed that Gas-mi R01 could downregulate the expression of inflammatory factors in the mice’s brain and inhibit the activation of the NF-κB signaling pathway.In summary,animal experiments are consistent with the results of cell experiments,indicating that Gas-mi R01 can regulate NF-κB signaling pathways while providing strong evidence for the anti-inflammatory effects of Gas-mi R01.The results of this paper lay a foundation for exploring the cross-kingdom regulation mechanism of G.elata mi RNAs and provide theoretical basis for the subsequent research of G.elata miRNAs.