Induction of Tetrodotoxin exposure to oxidative stress injury of BV-2 Cells
Objective To investigate the induction of tetrodotoxin(TTX)exposure to oxidative stress damage of microglia BV-2 in mice.Methods BV-2 was selected as the cell model for TTX exposure in vitro.Firstly,BV-2 cells were exposed to TTX at different concentrations(0,0.01,0.1,1,10,100 μmol·L-1)to observe the changes of cell morphology and vitality.CCK8 method was used to determine the combined exposure concentration of TTX.BV-2 cells were exposed using a mixture of veratridine(VTD)and ouabain(O)with TTX,and the types of ion channels on the BV-2 cell membrane were validated based on cell morphology and cell viability.The effects of TTX exposure on BV-2 cells were evaluated by determining lactate dehydro-genase(LDH)release,calcium ion fluorescence,reactive oxygen species(ROS)content and cell apoptosis rate after TTX expo-sure alone.Results Under experimental concentration exposure,100 μmol·L-1 TTX could change the morphology of BV-2 cells and inhibit cell viability.Under combined exposure,TTX could counteract the difference of sodium concentration in and outside the cell membrane caused by VTD and O mixture and delay the cell injury state.Compared with the control group,100 μmol·L-1 TTX could induce the change of cell membrane permeability and release 4.01 U/g LDH.The relative fluorescence in-tensity increased to 84.78%and 63.48%,respectively.The apoptosis rate increased by 2.9 times with significant difference.Conclusion Under the conditions of exposure concentration and time,TTX exposure can inhibit the proliferation of BV-2 cells,change the permeability of cell membrane,promote the accumulation of reactive oxygen species in cells,and induce oxidative stress injury of cells,resulting in cell apoptosis.
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