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OBJECTIVE：To study the e ffects of stilbene glycoside c（TSG）on phosphorylation of Thr 205，Ser404 sites of Tau protein in Aizheimer ’s disease （AD）model mice ，and to investigate the potential anti-AD mechanism of TSG. METHODS ：APP/ PS1/Tau three transgenes （3×Tg-AD）mice were randomly divided into model group ，positive control group （huperzine，0.15 mg/kg），TSG low-dose ，medium-dose and high-dose groups （0.033，0.1，0.3 g/kg），with 6 mice in each group. In addition ，6 C57BL/6J mice were chosen as normal control group. Administration groups were given relevant medicine intragastrically. Model group and normal control group were given equal volume of normal saline intragastrically ，once a day ，for consecutive 60 days. After last medication ，immunofluorescence staining was used to detect Tau protein and phosphorylated Tau protein （Thr205， Ser404 sites） distribution and expression in brain tissue of mice in each group. Western blotting assay was used to detect phosphorylated Tau protein （Thr205，Ser404 sites）expression level in brain tissue of mice in each group. RESULTS ：Compared with normal control group ，the expression of Tau protein，phosphorylated Tau protein （Thr205，Ser404 sites）in 729011126@qq.com the brain tissue of mice were increased in model group ，which were easy to aggregate and distributed more widely ；theirrelative expression were increased significantly （P＜0.01）. Results of Western blotting assay showed that the expression levels of phosphorylat ed Tau protein （Thr205，Ser404 sites）were increased significantly （P＜0.01）. Compared with model group ，the expression of Tau protein ，phosphorylated Tau protein （Thr205，Ser404 sites） in the brain tissue of mice were decreased in positive control group and TSG groups ；aggregation decreased，distribution narrowed and their relative expression were decreased significantly （P＜0.01）. Results of Western blotting assay showed that the expression levels of phosphorylated Tau protein （Thr205，Ser404 sites）were decreased significantly （P＜ 0.01）. Compared with positive control group ，There was no significant difference in the distribution of Tau protein ，phosphorylated Tau protein （Thr205，Ser404 sites）in the brain tissue of mice in TSG groups ；the relative expression were not statistically significant（P＞0.05）；but Western blotting assay showed the expression levels of phosphorylated Tau protein （Thr205 site）in TSG medium-dose and high-dose groups as well as the expression levels of phosphorylated Tau protein （Ser404 site）in TSG groups were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can play an anti-AD effect on AD model mice by down-regulating the expression of phosphorylated Tau protein （Thr205，Ser404 sites）in brain tissue.
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OBJECTIVE：To study the regulatory effects of stilbene glucosid e（TSG）on c-Jun N-terminal kinase （JNK）and protein phosphortase 2B（PP2B）in APP/PS1/Tau transgenic dementia （3×Tg-AD）mice，and to explore its potential mechanism of anti-Alzheimer’s disease （AD）. METHODS ：Totally 45 male 3×Tg-AD mice were randomly divided into model group ，positive control group （huperzine A ，0.15 mg/kg），TSG low-dose ，medium-dose and high-dose groups （0.033，0.1，0.3 g/kg），with 9 mice in each group. Another 9 normal male C 57BL/6J mice were included into normal control group. Administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 60 d. Normal control group and model group were given constant volume of normal saline intragastrically. After medication ，Morris water maze experiment was used to test the spatial learning and memory ability of mice in each group ；Nissl staining was used to observe the changes of Nissl bodies in cerebral cortex and hippocampus ；mRNA and protein expressions of JNK and PP 2B were detected by qRT-PCR and Western blotting assay. RESULTS：Compared with normal control group ，the escape latency was significantly prolonged （P＜0.01），the retention time of the original platform quadrant was significantly shortened （P＜ and the times of crossing the platform was significantly reduced in model group （P＜0.01）；the number of Nissl bodies in cerebral cortex and hippocampus was significantly 729011126@qq.com reduced，the staining was slight ；the relative expressions of JNK mRNA and protein were significantly increased （P＜ 0.01），and the relative expressi ons of PP 2B mRNA and protein were significantly decreased （P＜0.01）. Compared with model group ，the escape latency was significantly shortened in positive control group and TSG groups （P＜0.01）；the retention time of the original platform quadrant was significantly prolonged （P＜0.01）；the times of crossing the platform was significantly increased （P＜0.01）；the number of Nissl bodies in cerebral cortex and hippocampus was increased significantly ，the staining was heavy ；the relative expression of JNK protein was significantly decreased（P＜0.05 or P＜0.01），the relative expressions of PP 2B mRNA and protein were significantly increased （P＜0.01）， while the relative expression of JNK mRNA was significantly decreased in TSG high-dose group （P＜0.05）. CONCLUSIONS ：TSG can improve the learning and memory ability and neuronal damage of 3 × Tg-AD mice. The mechanism may be related to down-regulating the transcription and expression of protein kinase JNK ，up-regulating the transcription and expression of protein phosphatase PP 2B.
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OBJECTIVE： To observe the effects of stilbene glycosidec （TSG） on okadaic acid （OA）-induced Tau protein phosphorylation in NG108-15 cells， and to investigate the potential anti-Alzheimer’s disease （AD） mechanism of this compound. METHODS： AD model of NG108-15 cells was induced by OA. The survival rate of NG108-15 cells was observed by MTT assay after pretreated with low-dose， medium-dose and high-dose of TSG （50， 100， 200 μmol/L）. The apoptosis of NG108-15 cells was detected by AO/EB double fluorescence staining. The protein and mRNA expression of CDK5 and GSK3β， and the protein expression of Tau and p-Tau were detected by Western blotting assay and RT-PCR. The distribution of CDK5， GSK3β and Tau protein were detected by immunofluorescence. RESULTS： The normal morphology of NG108-15 cells was observed in normal control group， but CDK5， GSK3β and Tau protein were not found or few was found. Contracted or globular early apoptotic cells were observed in model gorup； the distribution of CDK5， GSK3β and Tau protein was increased， while survival rate of the cells was decreased； protein and mRNA expression of CDK5 and GSK3β as well as ratio of the relative expression of p-Tau to that of Tau （p-Tau/Tau） were all increased significantly （P＜0.05 or P＜0.01）. After pretreatment of TSG， the distribution of early apoptotic cells as well as CDK5， GSK3β and Tau protein were all decreased to some extent in administration groups， while survival rates of the cells were increased significantly. Protein expression of CDK5 and p-Tau/Tau in medium-dose group and high-dose group as well as mRNA expression of CDK5， protein and mRNA expression of GSK3β in administration group were decreased significantly （P＜0.05）. CONCLUSIONS： TSG can protect against AD model cells， the effects of which may be associated with improving survival rate of the cells， down-regulating the protein expression and gene transcription level of phosphokinase CDK5 and GSK3β， inhibiting Tau protein phosphorylation.
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Objective To observe the effect of Zhuang Jing mixture (ZJM ) on behavior of primary senile rat and monoamine neurotransmitters in it′s brain ,and to study its mechanism of anti‐aging .Methods Fifty senile female rates which average weight was(300 ± 20)g were randomized divided into 5 groups :blank control group ,low‐,mid‐and high‐dose group of ZJM ,positive control group ,with 10 rats in each group .After medication for 8 weeks ,Morris water maze test was used to evaluate the spatial learning and memory ability of primary senile rats .High performance liquid chromatography with fluorescence detection (HPLC‐FD) was used to detect the monoamine neurotransmitter content in rat′s cerebral cortex and hippocampus .Results Compared with blank control group ,high‐,middle‐dose group of ZJM and positive control group were improved the ability of learning and memory of pri‐mary senile rat ,as well as the cerebral cortex and hippocampus monoamine neurotransmitters levels .High‐dose group of ZJM had significant difference compared with positive control group in improving the ability of learning and memory(P<0 .05) ,and in im‐proving the cerebral cortex and hippocampus monoamine neurotransmitters(P<0 .05) .Conclusion ZJM could significantly improve the learning and memory abilities of primary senile rats ,and its mechanism may be related to adjust the monoamine neurotransmitter in brain .
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OBJECTIVE:To study the effects of LYS polysaccharides on learning & memory and monoamine neurotrans-mitter content in SAMP8 brain.METHODS:50 6-month-old SAMP8 mice were randomized to 5 groups(10 in each group):SAMP8 group,huperzine A control group,low-,mid-and high-dose groups of polysaccharides.Another 10 6-month-old SAMR1 mice were assigned to normal control group.After medication for 40 days,the learning and memory abilities of mice in each group were detected with Morris water maze method.The norepinephrine(NE),dopamine(DA) and 5-HT contents in brain were determined by HPLC.RESULTS:The learning and memory abilities of SAMP8 group decreased significantly,and the NE,DA and 5-HT contents in brain decreased significantly compared with normal group(P
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OBJECTIVE:To optimize the conditions of the extraction technique for compound Liuyuexue gran?ules.METHODS:The herbs in formulation were first extracted by ethanol percolating,then the dregs were decocted with water to be extracted once more.Taking oleanolic acid and total flavones as evaluation markers,the extraction technique in two-step process was evaluated by orthogonal design.RESULTS:The optimum condition of ethanol percolating process was that the crude herb powder was soaked by8-fold80%ethanol for24hours and the optimum condition of water decocting process was that the dregs were decocted for3times with12-fold water,1.5hour each time.CONCLUSION:The repeated experimental results are stable,and the contents of components for markers are high.The optimum conditions can be used for enlarged pro?duction.
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Aim To observe the effects of LYS polysaccharides on expressions of APP, PS1, PS2 and ApoE in SAMP8 mouse brain. Methods 50 6-month-old SAMP8 mice were used and divided randomly into 5 groups: SAMP8 untreated control group, huperzine A control group, low-, mid-and high-dose groups of polysaccharides, with 10 mice in each group. 10 6-month-old SAMR1 mice were used as normal control. After each group was treated by corresponding drug for 40 days continuously, the mRNA contents of APP, PS1, PS2 and ApoE in brain tissue were assayed by reverse transcription polymerase chain reaction.Results The low-, mid-and high-dose groups of polysaccharides could reduce the mRNA contents of APP, PS1 and PS2, which showed a dose-effect relationship in some degrees. But it could not affect the content of ApoE. Conclusion LYS polysaccharides could inhibit the expressions of APP, PS1, and PS2 in SAMP8 mouse brain.