Preliminary Study On MiRNA Of Neolamarckia Cadamba And Houttuynia Cordata

N.cadamba and H.cordata are widely distributed,and they have many functions such as antibacterial,anti-inflammatory and blood sugar lowering.At present,the research on N.cadamba and H.cordata has focused on the analysis of various crude extracts or natural chemical constituents.Up to now,there has been no report on the miRNAs of N.cadamba and H.cordata.Recent studies have shown that miRNAs in plants can be a new way to participate in the regulation of species across borders.In this study,the miRNAs of N.cadamba and H.cordata were sequenced by high-throughput sequencing analysis,and the number and types of miRNAs contained in N.cadamba and H.cordata were identified,and their target genes were predicted bioinformatically.Then,a mouse model was used to preliminarily explore the cross-kingdom regulation of miRNAs in the N.cadamba.Results of the study are as follows:1.11,285,965 and 13,637,514 sequencing raw reads were obtained from N.cadamba and H.cordata respectively.After removing low-quality data,6,935,542 and 3,728,613 clean reads were obtained respectively.Then,192 miRNAs(167 conserved miRNAs and 25 new miRNAs)and 193 miRNAs(163 conserved miRNAs and 30 new miRNAs)were obtained from the database and software,and the length distribution statistics were obtained.The miRNAs of N.cadamba mainly concentrated in 24 nt and H.cordata were mainly concentrated at 28 nt.miRNAs were randomly selected to verify high-throughput sequencing results using real-time PCR.2.mi Randa software was used to predict the target genes of the N.cadamba and H.cordata in the human genome,and KEGG analysis showed that both of them may be involved in some key genes of the insulin pathway,suggesting that miRNA may be used as a kind of Regulatory measures to participate in the rise of human blood sugar and reduce physiological processes.3.12 mice were divided into two groups,6 mice in each group.Experimental group received total RNA from N.cadamba orally,while control group received saline.The test kept on 5 weeks.Body weight,average daily gain and feed intake of mice in the experimental group were significantly higher than the control group(P<0.05).The body composition of the mice was found to be significantly different between the experimental group and the control group(P<0.05).The mice were dissected and found that the adipose tissue and subcutaneous adipose tissue in the experimental group were significantly higher than that in the control group(P<0.05).HE staining of epididymal adipose tissue showed that the size of adipocytes in the experimental group was significantly higher than that in the control group(P<0.05).There was no significant difference in serum high-density lipoprotein cholesterol and lowdensity lipoprotein cholesterol between two groups.Genes related to adipose tissue formation in the epididymis of mice were detected.And it was found that FAS,PPAR-? and CD36 in the experimental group were significantly higher than those in the control group(P<0.05).4.We transfected N.cadamba miR166e,miR395 a,novel-23 and novel-28 into 3T3-L1 cells,respectively,and found that mi R166 e can significantly increase the triglyceride content of 3T3-L1 cells(P<0.01),western blot results showed that mi R166 e could up-regulate the protein level of FAS,and q RT-PCR results also showed that the expression levels of FAS and PPAR-? genes increased significantly(P<0.05).In summary,this study is a preliminary exploration and analysis of miRNAs from N.cadamba and H.cordata.Both in vivo and invitro tests suggested miRNA from N.cadamba and H.cordata.may be able to regulate fat metabolism,which needs further confirmation.