ectly protected neurons via modulating microglia in microglia neuron co culture system As SCM 198 could effectively inhibit microglial KOS 953 over activation, we then ventured into how these SCM 198 or IBU pretreated microglia would interact with Inhibitors,Modulators,Libraries neurons. A co culture system was applied to investigate whether SCM 198 could protect neurons indirectly via directly suppressing overactivated microglia. LPS preactivated or SCM 198 or IBU pretreated BV 2 cells in inserts were washed twice with fresh DMEM medium to re move residual LPS, SCM 198 or IBU. These washed cells in inserts were then placed into wells containing primary neurons and were co cultured for 24 hours. Between inserts and wells, there is a semi permeable membrane which blocks the direct contact between neurons and microglia, but allows e change of molecules.
LPS preactivated BV 2 cells caused a decrease in neuronal viability Inhibitors,Modulators,Libraries which was re versed by SCM 198 or IBU pretreated microglia and 1 uM SCM 198 turned out to be the optimal dose 9. 984, P 0. 0006, Figure 6a. Accordingly, West ern blot showed that phosphorylation of ERK and tau in neurons were repressed by SCM 198 4. 27, P 0. 0026, Figure 6c. F 3. 40, P 0. 0150, Figure 6d, F 5. 599, P 0. 0069, Figure 6e. F 8. 544, P 0. 0001, Figure 6f, respectively indicating that SCM 198 could indirectly protect primary neurons through suppressing micro glial overactivation. Meanwhile, SCM 198 could also directly protect neurons from 20 uM AB1 40 induced neuronal death 7. 07, P 0. 0008, Figure 6g and LDH leakage 23. 41, P 0. 0001, Figure 6h.
SCM 198 ameliorated cognitive deficits of AB1 40 injected SD rats in MWM test To further e plore neuroprotective effects of SCM 198 in vivo, SD rats with bilateral intrahippocampal injections of aggregated Inhibitors,Modulators,Libraries AB1 40 were applied. As the e periment progressed, average escape latencies of all groups gradually decreased, with no significant differences observed from trial 1 to trail 3 2. 06, P 0. 085. F 0. 98, P 0. 440. Inhibitors,Modulators,Libraries F 1. 11, P 0. 3668, respectively, Figure 7a. Up to trial 4, a dramatically significant decrease of escape latency was observed in 60 mg kg SCM 198 treated group as compared with that of only AB1 40 treated group 4. 70, P 0. 0013, Figure 7a. In trial 5 and trial 6, rats administered with 30 mg kg SCM 198 also showed considerable cognitive improve ments 4. 10, P 0. 0032. F 4. 00, P 0. 0037, respectively, Figure 7a.
Time spent in the target quadrant was assessed during probe trial. Figure 7b showed Cilengitide that SCM 198 enhanced spatial memory of rats in a dose dependent manner 5. 44, P 0. 0004, Figure 7b. Two way repeated measures ANOVA analysis showed a significant effect of drug treat ment 8. 667, P 0. 0001 and trial effect 84. 80, P 0. 0001. Body weight remains normal and Brefeldin A mw no statistical differences were found in swimming speed of rats between groups throughout the e periment. DON, a first line inhibitor of acetyl cholinesterase and now clinically used for AD treatment, was used as the positive control. Taken togethe