Effects of Enterovirus 71 on Mitochondrial Dynamics in Human Glioma U251 Cells
Abstract
To investigate the effects of enterovirus 71 (EV71) on mitochondrial dynamics in human Glioma U251 cells. Methods The EV71 was replicated in Vero cells and the 50% tissue culture infective dose (TCID50) was calculated based on the Reed-Muench formula. After the U251 cells were infected with EV71, the cellular morphology was assessed through the light microscope. The mitochondrial morphology was detected by MitoTracker Deep Red staining under laser confocal microscopy and the mitochondrial ultrastructure was visualized by transmission electron microscopy. The expressions of mitochondrial fission proteins Drp1, p-Drp1 and fusion protein Opa1 were examined by Western blot. The level of ATP was measured by a commercial ATP assay kit. The generation of mitochondrial superoxide was detected by MitoSOX staining. Results The TCID50 of EV71 was 10－5.4/0.1 mL. Twenty-four or 48 h after EV71 infection, the U251 cells appeared shrunken, round and dead. The laser confocal microscopy and transmission electron microscopy images showed that the EV71 infection induced mitochondrial elongation and cristae damage. Moreover, Western blot analysis demonstrated that the protein expressions of Drp1 and Opa1 were downregulated at both 24 and 48 h after EV71 infection in U251 cells, companied with a significant increase in Drp1 phosphorylation at 48 h after infection (P<0.05). In addition, a decreased ATP level and elevated mitochondrial superoxide generation were observed in the EV71 infected group, as compared to the control group. Conclusion Our study demonstrated that infection with EV71 led to changes of mitochondrial morphology and dynamics in U251 cells, which may impair mitochondrial function and contribute to nervous system dysfunction.
Keywords: Enterovirus 71, Human glioma U251 cells, Mitochondrial dynamics
Full Text:
PDFReferences
SOLOMON T， LEWTHWAITE P， PERERA D， et al. Virology, epidemiology, pathogenesis, and control of enterovirus 71. Lancet Infect Dis，2010，10(11): 778–790.
KIM H J， HYEON J Y， HWANG S， et al. Epidemiology and virologic investigation of human enterovirus 71 infection in the Republic of Korea from 2007 to 2012: a nationwide cross-sectional study. BMC Infect Dis， 2016，16(1): 425.
MIZUTA K， AOKI Y， MATOBA Y， et al. Molecular epidemiology of enterovirus 71 strains isolated from children in Yamagata, Japan, between 1990 and 2013. J Med Microbiol，2014，63(10): 1356–1362.
GU Y Y， SHI K， YAO S， et al. Morphological characteristics of fatal pediatric hand, foot and mouth disease: a clinicopathological study with related receptors of EV71. Pathol Res Pract，2017，213(9): 1144–1151.
HORWOOD P F， ANDRONICO A， TARANTOLA A， et al. Seroepidemiology of human enterovirus 71 infection among Children, Cambodia. Emerg Infect Dis，2016，22(1): 92–95.
KHACHO M， SLACK R S. Mitochondrial dynamics in the regulation of neurogenesis:from development to the adult brain. Dev Dyn，2018， 247(1): 47–53.
BAEK S H， PARK S J， JEONG J I， et al. Inhibition of Drp1 ameliorates synaptic depression, Aβ deposition, and cognitive impairment in an Alzheimer's Disease Model. J Neurosci，2017，37(20): 5099–5110.
FILICHIA E， HOFFER B， QI X， et al. Inhibition of Drp1 mitochondrial translocation provides neural protection in dopaminergic system in a Parkinson's disease model induced by MPTP. Sci Rep， 2016， 6:32656[2019-09-05]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5020318/. doi:10.1038/srep32656.
WANG H， YI J， LI X， et al. ALS-associated mutation SOD1G93A leads to abnormal mitochondrial dynamics in osteocytes. Bone，2018，106: 126–138.
JOSHI A U， EBERT A E， HAILESELASSIE B， et al. Drp1/Fis1-mediated mitochondrial fragmentation leads to lysosomal dysfunction in cardiac models of Huntington's disease. J Mol Cell Cardiol，2018，127: 125–133.
RAMAKRISHNAN M A. Determination of 50% endpoint titer using a simple formula. World J Virol，2016，5(2): 85–86.
ZHOU H， ZHANG Y， HU S， et al. Melatonin protects cardiac microvasculature against ischemia/reperfusion injury via suppression of mitochondrial fission-VDAC1-HK2-mPTP-mitophagy axis. J Pineal Res， 2017，63(1): e12413 [2019-09-05]. https://doi.org/10.1111/jpi.12413.
JACKSON J G， ROBINSON M B. Regulation of mitochondrial dynamics in astrocytes: mechanisms, consequences, and unknowns. Glia，2018， 66(6): 1213–1234.
FLIPPO K H， STRACK S. Mitochondrial dynamics in neuronal injury, development and plasticity. J Cell Sci，2017，130(4): 671–681.
ZHU X， PERRY G， SMITH M A， et al. Abnormal mitochondrial dynamics in the pathogenesis of Alzheimer's disease. J Alzheimers Dis， 2013，33 Suppl 1: S253–S262.
BARBIER V， LANG D， VALOIS S， et al. Dengue virus induces mitochondrial elongation through impairment of Drp1-triggered mitochondrial fission. Virology，2017，500: 149–160.
KECK F， BROOKS-FAULCONER T， LARK T， et al. Altered mitochondrial dynamics as a consequence of Venezuelan Equine encephalitis virus infection. Virulence，2017，8(8): 1849–1866.
KIM S J， SYED G H， KHAN M， et al. Hepatitis C virus triggers mitochondrial fission and attenuates apoptosis to promote viral persistence. Proc Natl Acad Sci U S A，2014，111(17): 6413–6418.
YOU D G， LEE H R， KIM W K， et al. Hepatitis C virus p7 induces mitochondrial depolarization of isolated liver mitochondria. Mol Med Rep，2017，16(6): 9533–9538.
TEODOROF-DIEDRICH C， SPECTOR S A. Human immunodeficiency virus type 1 gp120 and tat induce mitochondrial fragmentation and incomplete mitophagy in human neurons. J Virol，2018，92(22): e00993–18[2019-09-08] . https://doi.org/10.1128/JVI.00993-18.
Refbacks
- There are currently no refbacks.
