This way, we could ensure that we would assess transcript profiles of the host cells under the same condition and time as assessment of antiviral activity and cytotoxicity for the same replicates.
At 72 hours post infection, antiviral activity and cytotoxicity of Hep at 125 µg/mL in 12-well plates were carried out at the same time as RNA extraction. SK-N-SH cells were infected with a clinical EV71 isolate followed by treatment with 125 µg/mL of Hep. The microarray analysis reported here was a large-scale microarray pilot study and thus further confirmatory experiments such as real time RT PCR and Western blotting would be warranted in order to confirm the mode of action of Hep implied here. There is no approved drug for prophylaxis of EV71-casued disease and discovering a molecular drug target(s) for EV71 infection would be beneficial. EV71 is a neurovirological virus that can cause severe and fatal CNS complications in infected patients. In conclusion, we propose that our microarray findings may suggest new directions for further studies on molecular targets of anti-EV71 activity of Hep. Furthermore, it should not be ignored that the Hep-caused affects seen in this microarray analysis may partially be attributed to a significant inhibitory effect of Hep on EV71 entry into the cells. In addition, Hep sometimes appeared to be harmful for cells, like the case in that Hep likely suppresses regulation of Metallothionein 1E. The most important genes where expression patterns were significantly changed by Hep include genes related to (i) cell growth, (ii) DNA repair and replication, (iii) cytoplasmic microfilaments and related apoptosis, (iv) regulation of energy metabolism and mitochondrion-related apoptosis, (v) cytoplasmic viral transport, (vi) transmembrane proteins and the related signaling pathways, (vii) phosphoribosyltransferase, (viii) transcription factors, (ix) G protein-coupled receptors and G protein-coupled receptors-interacting proteins, (x) remodeling actin filament assembly, (xi) chemokin receptors, and (xii) immunosuppression. Mostly, Hep appeared to modulate induction effect of EV71 on several genes in a way that may benefit the host cell and inhibit the viral infection. Overall, the findings of this study suggest several molecular targets by which Hep might exert its antiviral activity against clinical EV71 infection in neural cells.
Instead of stating a long list of gene functions and pathways, we tried to select for EV71-induced genes that were exclusively affected by antiviral activity of Hep through a multi-level comparison and characterization. Therefore, in this study we intended to gain insight into the cellular and molecular mechanisms of action of Hep against clinical EV71 infection in neural cells. We have previously shown that Heparin (Hep) significantly inhibited Enterovirus 71 (EV71) infection and binding in both Vero and a human neural cell line, SK-N-SH, in vitro. Microarrays analysis of anti-Enterovirus 71 activity of Heparin GEO help: Mouse over screen elements for information.
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