Abstract：Objective To explore the killing mechanism induced by Coxsackievirus A16（CV-A16）in primary muscle cells of gerbils，and to lay the foundations for elucidation the pathogenesis of CV-A16 and the further application of gerbil model.Methods The primary muscle cell model was established by digestion of trypsase/collagenase double enzyme hydrolysis. Primary muscle cells were infected by different dose of CV-A16 and the cell viability was detected by CCK-8 assays. Chromatin condensation and break were measured by Hoechst 33258 staining. The early and last stage of apoptosis cells were measured by AnnexinV/PI double staining. Expression changes of Caspase-3，Caspase-8，JNK and NF-κB pathway proteins were detected by Western Blot.Results The cell viability were 88.95% and 64.05% at groups of different multiplicity of infection（MOI=0.50 and 1.00），which was significantly different from those of the negative control group. The cell viability and multiplicity of infection were negative correlation（rs=-0.857，P=0.014）. The apoptosis rates were 7.2%，21.8% and 50.7% at MOI=0.01,0.10 and 1.00 groups，respectively. The apoptosis rate and MOI were positive correlation（rs=1.000，P<0.001）. When the primary cells were infected by CV-A16，cleavage of Caspase-3 and Caspase-8 were detected. Western Blot assays showed that the expression of NF-κB pathway proteins IκBα，p65 and p-p65 were reduced，which was different in enterovirus 71-infected cells. The JNK kinase was actived.Conclusion CV-A16 could induce apoptosis in primary muscle cells from gerbils.
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