The Effects Of TREK-1 Overexpression And Mailuoning Injection On Astrocytes And Glutamate Metabolism After Oxygen And Glucose Deprivation In Vitro

Objective : 1.To investigate the influence of TREK-1 overexpression and Mailuoning Injection on proliferation rate and glutamate uptake efficacy in primary cultured Astrocytes subjected to oxygen-glucose deprivation and the role of TREK-1on glutamate excitotoxicity.Methods: 1. The oxygen and glucose deprivation(OGD) was established as in vitro model of cerebral ischemia; The TREK-1 overexpression of astrocytes was made by DNA plasmid transfection; 2. Control group, TREK-1 overexpression group and Mailuoning Injection group were settsd.MTT was used to detect the cell survival rates at differet OGD time. Two methods was used to detect cellapoptosis rates:FITC/Probidium Iodide and Hoechst 33258. The about Glutamate oxidase Assay Kit was used to detect glutamate uptake rates and the expression rates of GLT-1, P2X7 R,GS, Caspase-3, TREK-1 mRNA were detected by Real-Time PCR.Results: 1. Using rat recombinant DNA plasmid could successfully establish TREK-1 channel overexpression model. 2. Under the normal condition, TREK-1overexpression and Mailuoning injection had no effects on cell viability of astrocytes.But after OGD, TREK-1 overexpression and Mailuoning injection could reduce the apoptosis rates. 3. Astrocytes’ Glumate uptake efficacy was increased after they subjected to OGD, and TREK-1 overexpression and Mailuoning injection could enhance this effect. 4. TREK-1 overexpression and Mailuoning injection could increase the mRNA expression of GLT-1, GS, TREK-1, reduce the mRNA content of Caspase-3 Where as they had no significant effect on P2X7 R expression under the condition of OGD.Conclusions:1. Using recombinant TREK-1 DNA plasmid could successfully transfected the rat astrocytes. 2 TREK-1 overexpression and Mailuoning injection could increase the astrocyte cell viability and enhance glutamate uptake of astrocytesubder oxygen glucose deprivation.