Abstract:Objective To explore the mechanism of death induced by EV71 virus in primary muscle cells of gerbils, and to lay foundations on the research of infection mechanism of EV71 virus. Methods The primary muscle cells were isolated and cultured by digestion of tyrisin/collagenase. Survival rates of primary muscle cells infected by EV71 virus were detected by CCK-8 assays. Changes of the nuclei in infected cells were observed by Hoechst 33258 staining. The redistribution of phosphatidylserine from the inner to the outer leaflet of the plasma membrane was measured by AnnexinV/PI double staining. Expression changes of proteins PARP-1, Caspase-3, Bcl-xl, Bcl-2 and p65 were detected by Western Blot. Results We established a primary muscle cells model in vitro successfully. Primary muscle cells form gerbils would be shrank, round and floated in the culture medium after infected by EV71 virus. Several events in apoptosis such as nuclear condensation, phosphatidylserine translocated to the outside of the plasma membrane, cleavage of apoptosis-related proteins PARP-1 and Caspase-3 were detected. Further studies showed that EV71 infection could not only activate the NF-κB pathway, but also suppress the apoptosis-suppressing proteins Bcl-xl and Bcl-2, leading apoptosis in primary muscle cells. Conclusion EV71 could induce apoptosis in primary muscle cells from gerbils.
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