Protective Actions Of Intravenous Anesthetics On Cultured PC12 Cells Impaired By Mimic Ischemia And Following Reperfusion

Objective 1. To evaluate the protective actions of intravenous anesthetics propofol, midazolam and ketamine, as well as combination of propofol with midazolam or ketamine on cultured PC 12 cells impaired by mimic ischemia and following reperfusion and the possible mechanisms. 2. To evaluate the protective actions of monoamine transmitter inhibitors reserpine and para-chlorophenylalanine (pCPA) on cultured PC 12 cells impaired by mimic ischemia and following reperfusion and the possible mechanisms, as well as the changes of these effects when these inhibitors are combined with propofol. Methods 1. Cellular culture: the nerve growth factor (NGF) differentiated PC 12 cells were planted in 96 or 6 well plates in the DMEM culture complete medium consisted of 5% heat-inactivated fetal calf serum and horse serum independently. Incubation was carried out at 37 in a humidified atmosphere with 5% CO2. 2. To detect the protective actions of propofol and midazolam on PC12 cells impaired by mimic ischemia and reperfusion(I/R), the cells in the 96 wells plate were allocated into control group (impaired by ischemia and reperfusion without any treatment), propofol-treating group (at dose of 12.4 mol-L-1) midazolam-treating groups (at dose of 3.3, 10, 30 mol-L-1), and propofol/midazolam-treating groups. The growth PC 12 cells, which were incubated in the 6 wells plate, were used to detect the changes of [Ca2+]i. The cells were allocated into control group, propofol-treating group (12.4 mol-L"1), midazolam-treating groups (3.3, 10 mol-L"1), and propofol/midazolam-treating group. 3. To detect the protective actions ofpropofol and ketamine on PC 12 cells impaired by mimic I/R, the cells in the 96 wells plate were allocated into control group, propofol-treating groups (12.4umol-L~'), ketamine-treating groups (3.6, 18, 90umol-L"1), and propofol/ketamine-treating groups. The growth PC 12 cells, which were incubated in the 6 wells plate, were used to detect the changes of [Ca2+]|. The cells were allocated into control group, propofol-treating group (IZ^umol-L"1), ketamine-treating groups (3.6, ISumol-L-1), and propofol/ketamine-treating group. 4. To detect the protective actions of propofol and reserpine on PC 12 cells impaired by mimic I/R, the cells in the 96 wells plate were allocated into control group, propofol-treating groups (12.4, 37.4, 112 mol-L-1), reserpine-treating groups (2.5, 10, 40umol-L"1), and propofol/reserpine-treating groups. The growth PC 12 cells, which were incubated in the 6 wells plate, were used to detect the changes of [Ca2+]j. The cells were allocated into control group, propofol-treating group (12.4, 37.4 mol-L-'), reserpine-treating group (40 mol-L-1) and propofol/reserpine-treating group. 5 To detect the protective actions of propofol and para-chlorophenylalanine (pCPA) on PC 12 wells impaired by mimic I/R, the cells in 96 wells plate were allocated into control group, propofol-treating groups (12.4, 37.4, 112umol-L~'), pCPA-treating groups (1.1, 3.3, 10 mol-L-1) and propofol/pCPA-treating groups. The growth PC12 cells, which were incubated in the 6 wells plate, were used to detect the changes of [Ca2+]j. The cells were allocated into control group, propofol-treating group (12.4, 37.4 mol-L-1), pCPA-treating group (10 mol-L-1) and propofol/pCPA-treating group. 6 We exposed the PC 12 cells to experimental oxygen-glucose deprivation culture medium in a humidified 95%N2, 5%CO2mixed gases at 37 to mimic ischemia impairment, and then incubated in DMEM complete culture medium at 37 in a humidified 5% CO2 atmosphere to mimic the following reperfusion. The degree of cellular impairment and the cellular viability were detected using LDH and MTT assays. Meanwhile, the change of [Ca2+]i was detected by Fura-2/AM fluorescence measure. Results 1. Compared with control group, the Asvonm values increased in all of the groups treated with propofol, midazolam and ketamine (p<0.05), which indicated increasing cell survival rates. Also, the protective actions of propofol were enhanced by midazolam and...