Effect of Coxsackievirus A16 on primary gerbil muscle cells and its mechanism
10.19485/j.cnki.issn2096-5087.2018.04.001
- VernacularTitle:CV-A16对长爪沙鼠肌肉细胞的杀伤机理研究
- Author:
Yi-Sheng SUN
1
;
Ping-Ping YAO
;
Zhang-Nyu YANG
;
Fang XU
;
Hang-Jing LU
;
Li-Zhi WU
;
Han-Ping ZHU
Author Information
1. 浙江省疾病预防控制中心
- Keywords:
Coxsackievirus A16;
Apoptosis;
Gerbil;
Molecular mechanism
- From:
Journal of Preventive Medicine
2018;30(4):325-328,333
- CountryChina
- Language:Chinese
-
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.
