The Effects Of Exosome-derived MiRNA-200b On Hydrogen Protecting The OGD/R-related Neuron Injury

Aims:By culturing primary rat hippocampal neural cells,we eatablished oxygenglucose deprivation/ reperfusion(OGD/R)model at 20? to mimic the DHCA-related neuron injuries,thus to explore the influence of exosome-derived mi R-200 family on the OGD/R-related neuron damage and the roles of hydrogen on the mi R-200 family.Providing certain basis for the further study on the exosome's mechannism on neural cell injury.Methods: Neuron-specific enolase(NSE)was used to identify the cultured primary rat hippocampal neural cells.Hippocampal cells were divided into 4 groups to isolate exosomes:(a)control group:NC-Exo;(b)hydrogen group: H2-Exo;(c)OGD/R group: OGD/R-Exo;(d)OGD/R+H2 group:OGD/R+H2-Exo.Electron microscopy,Western blotting and Nanoparticle Tracking Analysis(NTA)were used to identify the exosomes.Real-Time PCR analysis was used to determine the mi R-200 family in exosomes.Results: The exosomes were successfully obtained from the supernatant of cultured hippocampal cells.OGD/R increased the levels of mi R-200 a,mi R-200 b,mi R-200 c,mi R-141 and mi R-429 in exosomes,and it showed the greatest impact on mi R-200 b.60% hydrogen decreased the levels of exosomes mi R-200 family.We found no significant difference of mi R-200 family in NC-Exo and H2-Exo.Conclusions:Hydrogen reversed OGD/R-induced up-regulated exosome mi R-200 family in hippocampal cells.Aims:Objective to investigate the expression level of exsomal mi R-200 b and its significances in OGD/R-induced hippocampal cells injury,and further to study the effects of hydrogen on hippocampal-derived exosomes.Methods:Co-cultured hippocampal cells were divided into four groups:(A)control group;(B)hippocampal cells + OGD/R-Exo;(C)hippocampal cells +(OGD/R+H2-Exo);(D)hippocampal cells +(OGD/R-Exo)+antagomi R-200 b.Real-Time PCR analysis was used to determine the mi R-200 b in recipient cells.MTT method was used to analysis the recipient cells proliferation;DHE method was used to measure reactive oxygen species(ROS);Western blot and flow cytometry were conducted for apotosis.Results: The OGD/R-Exo resulted in an elevated level of mi R-200 b in recipient cells,which eventually leaded to inhibited proliferation,increased ROS,apoptosis in recipient hippocampal cells.Pre-transfection of hydrogen or antagomi R-200 b remarkably decreased the levels of mi R-200 b in recipient cells,with the injuries recovery.Conclusions: Exosomes played an important role in the secretion of hippocampal cells stimulated by OGD/R,which can induce neuron injuries by carrying mi R-200 b.The neuron cells protective effect of hydrogen was related to the down-regulation of exosomal mi R-200 b.