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.
MA Z, ZHA J, Yang J, et al. Epidemiological and genetic analysis of hand-foot-mouth disease by enterovirus A71 in Taizhou, P. R. China, between 2010 and 2013 [J]. J Med Virol, 2016. DOI:10.1002/jmv.24697.
[2]
LIU S L, PAN H, LIU P, et al. Comparative epidemiology and virology of fatal and nonfatal cases of hand, foot and mouth disease in mainland China from 2008 to 2014 [J]. Rev Med Virol, 2015, 25(2): 115-128.
[3]
WANG S M, LIU C C. Update of enterovirus 71 infection: epidemiology, pathogenesis and vaccine [J]. Expert Rev Anti Infect Ther, 2014, 12(4): 447-456.
[4]
CONG H, DU N, YANG Y, et al. Enterovirus 71 2B induces cell apoptosis by directly inducing the conformational activation of the proapoptotic protein Bax [J]. J Virol, 2016, 90(21): 9862-9877.
[5]
WANG C, ZHOU R, ZHANG Z, et al. Intrinsic apoptosis and proinflammatory cytokines regulated in human astrocytes infected with enterovirus 71 [J]. J Gen Virol, 2015, 96(10): 3010-3022.
[6]
DU H, YIN P, YANG X, et al. Enterovirus 71 2C protein inhibits NF-kappaB activation by binding to RelA(p65) [J]. Sci Rep, 2015(5):14302.
[7]
ZHANG H, LI F, PAN Z, et al. Activation of PI3K/Akt pathway limits JNK-mediated apoptosis during EV71 infection [J]. Virus Res, 2014(192):74-84.
[8]
LU J R, LU W W, LAI J Z, et al. Calcium flux and calpain-mediated activation of the apoptosis-inducing factor contribute to enterovirus 71-induced apoptosis [J]. J Gen Virol, 2013, 94(Pt 7): 1477-1485.
[9]
XU F, YAO P P, XIA Y, et al. Enterovirus 71 infection causes severe pulmonary lesions in gerbils, meriones unguiculatus, which can be prevented by passive immunization with specific antisera [J]. PLoS One, 2015, 10(3): e0119173.
[10]
YAO P P, QIAN L, XIA Y, et al. Enterovirus 71-induced neurological disorders in young gerbils, Meriones unguiculatus: development and application of a neurological disease model [J]. PLoS One, 2012, 7(12): e51996.
[11]
SOMOVA L I, GREGORY M A , Nxumalo E N. Mongolian gerbil (Meriones unguiculatus) as a model of cerebral infarction for testing new therapeutic agents [J]. Methods Find Exp Clin Pharmacol, 2000, 22(4): 203-210.
[12]
NOTO J M, ROMERO-GALLO J, PIAZUELO M B, et al. The Mongolian gerbil: a robust model of helicobacter pylori-induced gastric inflammation and cancer [J]. Methods Mol Biol, 2016(1422):263-280.
[13]
KIRAZ Y, ADAN A, YANDIM M K, et al. Major apoptotic mechanisms and genes involved in apoptosis [J]. Tumour Biol, 2016, 37(7): 8471-8486.
[14]
GOLDAR S, KHANIANI M S, DERAKHSHAN S M, et al. Molecular mechanisms of apoptosis and roles in cancer development and treatment [J]. Asian Pac J Cancer Prev, 2015, 16(6): 2129-2144.
[15]
GIBSON C J, DAVIDS M S. B[J]. Clin Cancer Res, 2015, 21(22): 5021-5029.
VARBLE A J, RIED C D, HAMMOND W J, et al. The vesicular stomatitis virus matrix protein inhibits NF-kappaB activation in mouse L929 cells [J]. Virology, 2016(499):99-104.
