Research On Regulation Of Trichinella Spriralis Excretory-secretory Products By MicroRNA Tsp-Novel-46

microRNAs （miRNAs） could bind hundreds of different target mRNAs, whichcould be degradated or inhibited in the process of translation.It is one of the mostimportant gene transcription regulation factor in post transcriptional regulation ofgene expression and. regulations. These regulation comes in genome function of manyimportant levels, including the chromatin structure, transcription, RNA stability andtranslation, acompany with silence after gene transcription, cell growth, diseases andcontrol, genomic DNA and histone methylation, chromosome heterochromatinformation and stability, and regulating genome rearrangement.Studies have shownthat it is some highly expressed micrornas in human/mammals drives vital role.Helminth microRNA could not only participate in the development of the parasite,physiological metabolism and sexual differentiation process of life, but also intransferring information with host and regulating the host immune response toparasitic infections. Existing research shows that, for example, Trypanosoma bruceimicroRNAs family can induce immune protection and evade the host immune systemfrom attacking by regulating mRNAof the surface glycoprotein （VSG） antigen.Mature and the application of high-throughput sequencing technologies give agrowth in enhancing number of new miRNAs in parasite. My laboratory has gainedTrichinella spiralis relatively high expression of mirnas in the transcriptome study. Tounlock the information of Trichinella spiralis miRNA function, we use bioinformaticstechnology for mass screening the principle gene tatgets. Our study found thattsp-Novel-46targets gene could encode proteins with special physiological andbiochemical characteristics. These target proteins contains a lot of classical andnonclassical secretory proteins secreted protein, which reveals that microRNAtsp-Novel-46has a biological basis on regulation of secretion proteins.Trichinellaspiralis sixth day adult （AD6） is in the intestinal phase to attack the host. Thecomposition of Trichinella spiralis excretory secretions contain large number ofbiologically active substances and secreted proteins, including enolase,70kDa heatshock protein （HSP70）, FDPALD, glyceraldehyde-3-phosphate dehydrogenase,Linsuanganyousuan kinase, Glutathione transferase, TRX peroxidase ect., which could drive specific and nonspecific immune suppression and evade the host immuneresponse.This reveals tsp-Novel-46may regulate the expression of a variety of ESproducts, and play an important role in the process of immune escape.Thus, we collected the cultivation which contain ES proteins of AD6adult andnewborne larvae. We optimize and establish the2D electrophoresis （2-DE） system. ofexcretory secretory for deep understanding the tsp-Novel-46regulation effects. Wecollected Trichinella spiralis AD6adults and transfected tsp-Novel-46to the wormsby soaking in the medium of culture solution. And seek out the differencial ES proteinspot by comparing the2D-DIGE maps between transfection.group and normal one.The differentially expressed proteins were identified by LC-MS/MS. Two dimensionalgel electrophoresis proteomics results reveal that microRNA tsp-Novel-46Interference induce the differences expression level of proteins in ES products, whichmeans tsp-Novel-46has a regulation effect on ES products by promote or restrict thetargets.The identification results show that these proteins come from various ofproteins, including serine proteinases, grain protein and some unknown functionputative protein, which are all secreted proteins. Further analysis showed thatmicroRNA tsp-Novel-46could bind3’UTR in mRNA of Tsp₀4734, Tsp₀4685andTsp₀4680. According to the result of the experiment analysis, we suggestmicroRNA tsp-Novel-46may directly inhibit or catalytic protein expression asupstream regulatory factors, or associated with unique style "cascade" to adjust themetabolic process of ES product. It is the mutual coordination and balance betweenpath regulated by tsp-Novel-46,proteins might best activity and function.Proteins are basic materials of life activities,which plays vital effects onmaintaining the life activities.ES proteomics is one of the most important omics inTrichinella species.Components in Trichinella spiralis AD6ES proteins exposesdirectly to the host’s immune system, which are the main target antigens in inducingthe immune response. The changes of some antigen expression could affects thebody’s immune signaling pathway, and are connected with mechanism in evadingthe host immune response, triggerring specific and nonspecific immuneinhibition.Combining with bioinformatics method, two-dimensional electrophoresisand RNA transfection technique, we provide an important clue for the illustration oftsp-Novel-46biological function and the intestine invasion mechanism of Trichinellaspiralis, making for further elucidating the Trichinella spiralis in innate and adaptiveimmunity in the adjustment mechanism At present, Trichinella spiralis miRNAs is not yet clear, this study of Trichinellaspiralis miRNA has an innovation in the research on ES regulation,which could bringoverall strategy to the study of individual Trichinella spiralis miRNAtarget genes.