Lithium Chloride Reduce The Apoptosis Of PC12Cells, Induced By Anoxia In The Expression Of α5β1Dependent Manner

Objective:To explore the effect of Lithium chloride on the apoptosis and survival of PC12cells induced by anoxia，and confirm the neuroprotective effect of Lithium chloride invitro；For conducting the better used of Lithium chloride in the treatment ofischemia-hypoxia disease in clinical,providing theortical basis and research direction.Methods:PC12Cell Strain was provided from cell bank of Chinese Academy of SciencesShanghai Branch,incubated in a mixture of DMEM with10%FBS at37°C in5%CO2in a humidified atmosphere, and harvested just as reaching approx.70-80%confluency for experiment.After PC12exposed to anoxia for0h,12h,24h and36h, apoptosis was detectedby Annexin-v FITC/PI double-dyed flow cytometry,the group of0h as the controlgroup,12h,24h and48h as experimental groups. To observe the peak time ofapoptosis for post-experiment.Cultured PC12cells were divided into4groups with different culture manner:the control group,anoxia group,lithium chloride intervene anoxia group and lithiumchloride alone group. The rate of apoptosis was detected by Annexin-v FITC/PIdouble-dyed flow cytometry, viability of PC12cells were detected by CAM LiveCell dying;the expression of integrin α5、β1mRNA was analyzed by RT-PCR.Results：The percentage of apoptosis and mortality at the different time point of anoxia(12h,24h and36h) were higher than those of the control group, peaking at24hour.[23.12±0.048vs4.11±0.081,34.91±0.104vs4.11±0.081,36.58±0.092vs4.11±0.081,P＜0.05];But the36h group is a slight higher than24h group,but thedifference was not statistically significant,[34.91±0.104vs36.58±0.092,P>0.05].There were significantly increase in the rate of apoptosis,[34.91±0.117vs3.11±0.059；P＜0.05],and the expression of Caspase-3mRNA, but decrease in viability, [17.71±0.031vs90.11±0.035；P＜0.05],and the expression of integrin α5、β1mRNAinduced by anoxia of PC12; Pretreated with Lithium chloride(10mmol/l)30minbefore anoxia significantly prevent the increase of anoxia-induced apoptosis and theexpression of Caspase-3mRNA of PC12, also there were significantly higher inviability and the expression of integrin α5、β1mRNA of PC12against anoxiatreatment decrease when pretreated with Lithium chloride.Conclusion：Lithium chloride protected PC12cells against anoxia-induced apoptosis,whichwas maybe correlated with the inhibition of Lithium chloride on the decrease of theexpression of Integrin α5、β1of PC12cells induced by anoxia.