Spinal Cord Injury Affects Gene Expression ofTransmembrane Proteins in Tissue and Extracellular VesicleRelease in Blood: In Silico and In Vivo Analysis

Objective: Spinal cord injury (SCI) can disrupt membranetransmission by affecting channels or neurotransmitter release.This s tudy aimed to explore gene expression changes underlyingSCI in rats by bioinformatics approaches and confirm inRNA and protein levels.Materials and Methods: The differentially expressed genes(DEGs) in acute and subacute SCI were screened based onGSE۴۶۴, GSE۴۵۰۰۶, GSE۴۶۹۸۸, GSE۲۵۹۹ microarray datadownloaded from Gene Expression Omnibus. Transmembraneproteins of DEGs were recognized using the Uniprot and transmembranehelices prediction (TMHMM) methods. Model ofSCI was es tablished through a weight dropping procedure inrat. To confirm SCI model hematoxylin and eosin (H&E) s tainingwere performed. Total mRNA was extracted from spinalcord tissues. The RNA expression profile of some of the significantlychanged genes in pervious part has been confirmedby real-time polymerase chain reaction (RT-PCR). Blood wascollected from rats before sacrificing. EVs are isolated by highspeedcentrifugation (۲۰ Kg) from plasma. For assessment ofprotein expression of CD۹, CD۶۳ and Cxcr۴ proteins wes ternblotting were used.Results: Based on bioinformatics analysis, we found a set ofcommon genes in both phases that encode transmembrane proteins.Kcna۱, Scn۱a, Grm۱, Cnr۱ and Nrg۱ are downregulated;and Olr۱, Enpp۳, Cd۶۳, Ptprc, Cd۵۳, Adcy۴ Adcy۴, and Cxcr۴are upregulated. H&E demons trated tissue damage as well asneuronal degeneration. Following by RT-PCR for tissue, weshowed significant upregulation in Grm۱, Nrg۱, CD۶۳, Enpp۳,and Cxcr۴ between the acute and control groups and downregulationin Enpp۳ between acute and subacute groups. ConsideringCD۶۳ as an extracellular vesicle marker, we examined theprotein expression of CD۹ and CD۶۳ plasma derived-EV. CD۹has significant expression between acute and control groups.We also demons trate no significant CD۶۳ and Cxcr۴ expressionbetween groups.Conclusion: Our results provide novel insight into the relationshipbetween membrane protein expression and SCI. Also, resultsshow that SCI affected EV release in the blood which canhelp enlarge s trategies to enhance recovery following SCI.

کلیدواژه ها:

Bioinformatics ، Membrane Proteins ، Spinal Cord Injury