ABSTRACT
<p><b>OBJECTIVE</b>To investigate whether sodium valproate (VPA) directly regulates the activity of Ankyrin G(AnkG) promoter in vitro.</p><p><b>METHODS</b>The mouse AnkG promoter sequence was identified by comparing both human and mouse AnkG promoter sequences. The promoter was amplified from C57BL/6 mouse genome DNA and cloned into pGL3 Luciferase reporter vector. The Luciferase activity was detected in N2a and 293T cells and then treated with 0,0.5, and 1 mmol/L VPA for 12 h. The transcription activity of AnkG promoter in cells and the activity of VPA-treated Luciferase reporter vector in cells were detected using dual Luciferase reporter assay.</p><p><b>RESULTS</b>The AnkG promoter clone and its expression vector were successfully established, as confirmed by enzyme digestion and sequencing. The AnkG promoter showed high transcription activity in both N2a and 293T cells. The Luciferase activity was significantly induced following 0.5 mmol/L VPA treatment in both N2a and 293T cells. CONCLUSIONS VPA can up-regulate the AnkG expression via directly increasing its transcription activity. Thus, the in vivo AnkG expression may be directly regulated by the VPA at transcriptional level.</p>
Subject(s)
Animals , Humans , Mice , Ankyrins , Cell Line , Genetic Vectors , Luciferases , Mice, Inbred C57BL , Promoter Regions, Genetic , Up-Regulation , Valproic AcidABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of GETO on expression of PSD-95 and Shank-1 protein in postsynaptic dense zone in Alzheimer disease (AD) model rats.</p><p><b>METHODS</b>The AD model rats were established by beta-amyloid protein (Abeta(1-42)) injection into hippocampus CA1 zone. They were assigned into the model group, the donepezil treated group and the GETO treated group, besides, a normal group was set for control. Four weeks after modeling, Morris water maze test was applied to determine the learning and memory function of the AD rats, the number of PSD-95 and Shank-1 protein positive neuron as well as the optical density (OD) in post-synaptic dense zone of hippocampus CA1 area were determined by using immuno-histochemical stain and computerized graphic analysis techniques.</p><p><b>RESULTS</b>Morris water maze test showed that the mean escape latent period (MELP) of the model rats obviously prolonged than that of the normal rats, and the times of traversing flat roof obviously decreased (P < 0.01), while in the model rats after being treated by GETO, the two parameters were significantly shortened and increased respectively (P < 0.01), reaching the level insignificantly different to those in the donepezil group and the normal group. Immunohistochemical test showed that the number of positive stained neuron of PSD-95 and Shank-1 in hippocampus CA1 zone in the model group was significantly different to those in the normal group (P < 0.01), while in the GETO group those indexes were insignificantly different to those in the donepezil group and the normal group (P > 0.05), but showed a significant difference when compared with those in the model group (P < 0.05).</p><p><b>CONCLUSION</b>GETO can obviously improve the function of learning and memory of AD rats induced by Abeta(1-42), and the mechanism may be associated with its actions in improving expressions of PSD-95 and Shank-1 protein in hippocampus CA1 zone, and recovering the structure and function of synapse and enhancing its plasticity.</p>
Subject(s)
Animals , Male , Rats , Adaptor Proteins, Signal Transducing , Metabolism , Alzheimer Disease , Drug Therapy , Metabolism , Drugs, Chinese Herbal , Therapeutic Uses , Hippocampus , Metabolism , Maze Learning , Nerve Tissue Proteins , Metabolism , Phytotherapy , Rats, Sprague-Dawley , Synapses , Metabolism , Synaptic TransmissionABSTRACT
Alzheimer's disease (AD) is a common disease in elder people. Its incidence rate is about 5% in people above 60 years old. It has become an important factor that seriously impacts the development of families and society, and caused wildly attention all over the world. In this article, we discuss the mechanisms of AD in four aspects and put forward the strategies of drug therapy.
Subject(s)
Aged , Female , Humans , Male , Alzheimer Disease , Drug Therapy , Amyloid beta-Peptides , Metabolism , Amyloid beta-Protein Precursor , Metabolism , Apoptosis , Cholinesterase Inhibitors , Therapeutic Uses , Nerve Growth Factor , Metabolism , Neurofibrils , Pathology , Nootropic Agents , Therapeutic Uses , Plaque, Amyloid , Metabolism , Synapses , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Lishi No.5 formula on the growth of human neuroblastoma cell line SY5Y and find out the most effective drug dose.</p><p><b>METHOD</b>SY5Y cells were administrated by Lishi No.5 formula in different concentration. MTT metabolic rate was measured as the cell survival rate, then the dose-effect curve was made. LDH leakage rate, axonal length and area of cell body were used as the indicators.</p><p><b>RESULT</b>In 0.125-0.75 g x L(-1) Lishi No. 5 formula could increase the cell survival rate and MTT metabolic rate, decreased LDH leakage rate, and enhance axonal length and area of cell body, compared with control group.</p><p><b>CONCLUSION</b>Lishi No.5 formula has the neurotrophic effect and can induce the survival of neuron.</p>
Subject(s)
Humans , Cell Line, Tumor , Cell Proliferation , Cell Survival , Dose-Response Relationship, Drug , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , L-Lactate Dehydrogenase , Metabolism , Neuroblastoma , Metabolism , Pathology , Plants, Medicinal , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of Jiunaoyizhi capsul on the signal transduction pathway of SY5Y cell lines and explore the mechanism of the function of the capsul's enhancing neuronal growth.</p><p><b>METHOD</b>Human neuroblastoma was used as cell models and they were divided into control group and experimental group. Supernatant of cell lysate was taken and immunoprecipitation was done with antibodies to proteins related to signal transduction pathway, and the immunoprecipitates were analyzed by Western blotting.</p><p><b>RESULT</b>After treatment with Jiunaoyizhi capsul, expression of Akt/PKB, CREB, P-CREB was clearly increased and expression of cytochrome C decreased more than the control group.</p><p><b>CONCLUSION</b>Jiunaoyizhi capsul can promote expression of some proteins related with signal transduction pathway in SY5Y cell lines. Mechanism of Jiunaoyizhi capsul's enhancing neuronal growth is relevant to expression of some proteins in signal transduction pathway.</p>
Subject(s)
Humans , Cell Line, Tumor , Cell Proliferation , Cyclic AMP Response Element-Binding Protein , Metabolism , Cytochromes c , Metabolism , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Neuroblastoma , Metabolism , Pathology , Neurons , Plants, Medicinal , Chemistry , Signal TransductionABSTRACT
In Alzheimer's disease there is obvious evidence of insulin resistance in the brain. Thiazolidinediones,a kind of insulin sensitizer,not only improves insulin sensitivity,but also decreases inflammation,promotes release and clearance of?-amyloid protein,all are beneficial to the improvement of memory.