The Research On The Expression Of LCN-2 In Adenosine Preconditioning On Astrocytes After Oxygen-Glucose Deprivation And Reperfusion

Objective:Discussed the influence of adenosine preconditioning to the astrocytes which under the condition of Oxygen glucose deprivation/reoxygenation and the expression of lipocaline-2(LCN-2)in the medium,then to reveal the protection of adenosine in nerve cells and its clinical application to analyze the feasibility of cerebrovascular diseases.Methods:Primarily cultured the astrocytes of the Sprague-Dawley Rats in vitro,which were newborn within 24 hours.And then divided them into adenosine preconditioning control group(Ado-treated control),the control group(control),the experimental group(Ado/R)and model group(OGD/R)randomly,the experimental group and model group experienced the oxygen glucose deprivation/re-oxygenation,to establish the model of cerebral ischemia-reperfusion injury,the adenosine preconditioning control group and experimental group were experienced the adenosine preconditioning.24 hours’ later,inspect each group’s cell vitality by MTT,and to detect each group’s concentration of LDH by Elisa kits,and detect each group’s relative expression of lipocaline-2 by immunofluorescence technique and western blot test.Results:1.Light microscope:After oxygen-glucose deprivation and reoxygenation,the adenosine preconditioning control group and the control groups’ astrocytes grown well andthey didn’t have significant difference compared with before;The volume of the model group’s astrocytes were edema due to large shape from irregular shape to the fusiform,and the neurites changed obviously,scattered broken in the vision;The cells in the experimental group had a mildly edema change,but the change magnitude of the overall lower than that of the model group.2.MTT method was used to detect cell activity:After the oxygen-glucose deprivation and reoxygenation,model group’s cell activity decreased significantly compared with the control group,the OD value decreased significantly compared with the model group(P<0.05),the experimental group showed a higher cytoactive compared with the modle group,it’s show that adenosine preconditioning has a significant effect on astrocytes’ cytoactive(P<0.05).There is no significant difference between adenosine preconditioning control group and the control group(P>0.05).3.The concentration of LDH detected by ELISA kit:After oxygen-glucose deprivation / reoxygenation,the concentration of all groups’ LDH showed that:The concentration of lactic dehydrogenase in the model group was significantly higher than that of control group(P < 0.05),it release a large number of LDH;the concentration of the experimental group’s dehydrogenase(LDH)decreased significantly compared to the model group(P < 0.05),it indicate that the release of LDH decreased significantly and the cells’ damage was lighter than the model group.The concentration of dehydrogenase(LDH)between adenosine preconditioning control group and the control group had no significant difference(P>0.05).4,Immunofluorescence of LCN-2 showed that:The control group’s fluorescence signal was the weakest both on the cell membrane and in the cytoplasm;After the oxygen-glucose deprivation and reoxygenation,the expression of fluorescence intensity of model group’s astrocyte cells’ membrane and cytoplasm was significantly enhanced compared with the model group,whereas the astrocyte cells’ fluorescence intensity of membrane and cytoplasmic in experimental group decreased significantly compared with the control group.There is no significantdifference between adenosine preconditioning control group and the control group.5.The expression of LCN-2 in cells liquid :After oxygen-glucose deprivation and reoxygenation,in the four groups,the expression of lipocaline-2 in model group was the highest,and the experimental group was lower to it,and the control group was the lowest,the comparisons between either two groups have significant difference(P < 0.05).There was no significant difference between adenosine preconditioning control group and the control group(P>0.05).Conclusion:1.By detecting the activity of astrocytes and the content of LDH in the cells of each group,it show that adenosine can significantly reduce the damage caused by oxygen-glucose deprivation and reoxygenation;2.The release of LCN-2 is significantly correlated with the cell damage induced by oxygen-glucose deprivation and reoxygenation,adenosine can significantly improve the cellular damage caused by LCN-2 as a kind of inflammatory recruitment factor.