| Aim: In order to probe the mechanisms of compound 9714 for senile dementia and to find the effective targets for the acting of compound 9714 and to study the signal transduction pathways of compound 9714 at cellular level.Methods: The ICR mouse models of senile dementia induced by scopolamine were divided into several groups : control group, model control group, hydergine group, nimodipine group, 15mg/kg 9714 group,30mg/kg 9714 group,60mg/kg 9714 group. Detecting the effect of 9714 by the means of Morris (Morris water maze test) and Step Down Test . Proteins were extracted from the hippocampi and cortex of cerebrum .The expressions of M-type and N-type of acetylcholinergic receptors were measured by Western-Blotting . SHSY5Y cells , SK-N-SH cells and PC 12 cells were incubated in different concentration of 9714 or gin- seng saponin Rgl,Rbl and Re . And then decide the effect of 9714. Results:1. The compound 9714(15mg/kg,30mg/kg,60mg/kg) can improve the abilities of learning and memory in scopolamine induced senile dementia. 60mg/kg 9714 can shorten reactive period and prolong latent period in learning test of the Morris water escape task. 30mg/kg 9714 group or 60mg/kg 9714 group can cut down the time spent in the platform's quadrant and reduce the times of mistake.2. In the hippocampi, the quantity of Mi-type acetylcholinergic receptors protein in model control group was declined significantly to that in control group.But in hydergine group and nimodipine group, the quantity of MI-type receptors proteins were improved lightly to that in model control group .And in 9714 groups, the quantity of MI-type receptors proteins were improved significantly to that in model control group.And there were dose-effect relationship. The quantity of a,4-type receptors proteins in hydergine group and nimodipine group were similar, and both of them exceed that in model control group, a 7-type receptors protein in model control group was declined significantly to that in control group. But there have no difference between hydergine group and model control group. Nimodipine group and 9714 groups were both beyond the model control group in the quantity of receptors proteins . The most notable phenomenon is that a 7 and a4-type receptors were remarkably increased in 9714 groups and still with distinct dose-depend-effect. In the hippocampi, A -type receptors expression showed no difference in those groups. In thecortex, the quantity of a4-type receptors protein similar with its of a 7-type receptors proteins were higher in 9714 groups (30mg/ kg ,60mg/kg) than that in the model control group (P<0.05) .In contrary to the model control group there was no difference only in hydergine group whereas the significant difference in other groups about Mi-type and A -type receptors proteins (P<0. 05) .3.Neuron-like Cells were incubated in 9714 medium for 48h and then be observed .The cell bodies became larger and the cell projections were extended .The number of cells were increased distinctly and the cells activity were enhanced .The orders of effective concentration of 9714 to neuron-like cells are as follows: 80uM>40uM>20uM , SK-N-SH cells were more sensitive to 9714 than SHSY5Y cells and PC 12 cells were. SHSY5Y cells incubated both in 5% serum and 9714 (10uM,20uM) for 48h were significantly different to control groups in absence of 9714.Cells viability were measured using MTT assay test upon incubation for 48h with different 9714 concentrations . MTT assay data of SHSY5Y cells and SK-N-SH cells treated by 9714 significantly increased versus control groups among the effective concentrations in the range of 2.5uM~40uM (P<0.01) . But cells death can be distinctly detected in concentration of 80uM of 9714. Dose-dependent effect of 9714 (5uM~100uM) on PC 12 cells was significantly different to control groups in absence of 9714 (P<0.01) .PC 12 cells incubated with differentconcentration of 9714 will increase the quantity of a4-type,a7-type and MI-type receptors proteins in opposition to the control groups. Conclusions: The compound...
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