Metabonomicsanalysis Of Oligodendrogliomacells Infected With Bornadisease Virus

Background：Borne disease virus (BDV) is an enveloped,nonsegmented,negative-,single-stranded RNAvirus, BDV is a highly neurotropic virus. Alow level replication and transcription of the BDV genome take place in theinfected cells, to keep persistently infects. BDV does not establish acytolytic and obvious pathology but can lead to dysfunction of host cell. Thisvirus have a broad range of host, it can infect all warm-blooded animals,lead to Borne disease (BD). Recent years, extensive Epidemiologicalstudies show that BDV is related to human mental disorders besides.Themechanism underlying BDV pathogenesis is not well understood until now，There are a variety of hypotheses: immune response hypotheses,neurotransmitter hypotheses，neural plasticity hypotheses，neurotrophicfactor and Signal transduction et al. But the influence of BDV infection onhost metabolism is poorly known. Pletnikov MV et al reported that themetabolites such as NA, dopamine etc, are abnormal in BDV infected rat.We speculated that there is abnormal cell metabolism in the process of viralinfection，study the changes of these metabolites, is helpful to clarify themechanism of BDV chronic persistent infection. Objective:To get the broad spectrum metabolites of BDV-infected OL cell.Compare the differential metabolites between control and BDV-infectedhuman oligodendroglioma cells.Explain the pathophysiological mechanismsthat cause changes in metabolite, and provide basal differential metabolitesfor further BDV-induced pathogenesis exploration.Methods:Extract BDV virus liquid from cells which was persistent infectionwith BDV, reinfection OL cells with the virus and detection if successfulinfection by RT-PCR, immunofluorescence, and WB. liquid Intracellularsamples were collected from control and BDV-infected humanoligodendroglioma cells after14days post-infection.1H nuclear magneticresonance (1H NMR)-based metabonomics was used to characterizemetabolic profiles of both two groups. We compared the concentrations ofintracellular metabolites between both groups. Finally, we performed Partialleast squares discriminant analysis(PLS-DA).Results:Successfully extracted BDV virus liquid from virus-infected cells, andsuccessfully used the virus back OL infected cells. Fourteen samples fromcontrol and BDV-infected cells were analyzed. We quantified21metabolitesincluding amino-acids, dicarboxylic acids and organic acids. There aresignificant differences between the two groups: in BDV-infected groups pyruvate concentrations (p=0) are significantly lower than control groups.While concentration of glutamine higher (p=0), and those of glutamatetrended (p=0) and aspartate (p=0) were higher inthe BDV-infected groups.PLS-DA demonstrated that the pattern of analyzed metabolitesdiscriminated between groups.Conclusion:1H NMR-based metabonomics could be a simple, useful and low costmethod to explore the potential pathogenesis of virus infection. The resultsindicate BDV infection mainly affecting the metabolism of amino acids andthe glycolytic pathway. Glutamic salts significant increase in cells infectedby Virus may may explain the exciting symptoms of host which caused byBDV infection.