斑马鱼AD模型,阿尔茨海默氏病 z-bo 2020-09-28 577

　　AD (Alzheimer's disease) is a complex neurodegenerative disease that usually leads to cognitive decline and is the main cause of Alzheimer's disease. There are many in vitro and in vivo experiments that can prove that AD is related to decreased acetylcholine levels, peroxidation, increased levels of metal ions, and Aβ (amyloid β) accumulation, but the cause of AD is still unclear. Do not. In order to screen effective anti-AD drugs, an experimental animal model that can simulate the pathological process of AD must be established. For a long time, the mouse screening system has dominated the research of Alzheimer's disease. In recent years, the zebrafish model has received extensive attention from scholars and has been widely used in experiments due to its cost-effectiveness. For example, Paquet et al. A transgenic zebrafish model of tau protein was developed to discover compounds that phosphorylate tau protein. In addition, the zebrafish model can fully recognize GSK3β inhibitors, such as the therapeutic agent GSK3 that targets AD, which can reduce the brain of zebrafish fetuses. However, most zebrafish AD models are recombinant species, which are very time-consuming and expensive. Therefore, there is an urgent need for new zebrafish models that can quickly screen anti-AD drugs.

　　Aluminum chloride is a standard chemical used to induce AD models in rodents. Aluminum chloride can slow the swimming speed of zebrafish for 6-8 months. This is very similar to the dyskinesia phenomenon in AD patients. Therefore, aluminum chloride can be used to guide the creation of new zebrafish AD models. The reason for choosing 3-5 days after fertilization instead of 6-8 months of adulthood as described above is to better perform high-throughput drug screening and reduce costs. The model was validated by six antiretroviral therapies with multiple mechanisms of action or clinically or commercially available, and used it as a quantitative indicator to reflect the recovery rate (DRR) and response ability after light changes. (RE) is used to assess effectiveness. The results show that RE can be used as a reliable and convenient indicator for evaluating six anti-AD drugs. DRR is a measure of riboflavin, thioflavin T (ThT) and flurbiprofen. (Flurbiprofen) and AM-117. However, it does not include memantin (memantin) and rosiglitazone (rosiglitazone). We recommend using the new Zebrafish AD model in a practical system to screen new anti-AD compounds with high throughput.

　　Ibastigmin-acetylcholinesterase inhibitor

　　Ivastigmin is one of the five acetylcholinesterase inhibitors approved for the treatment of AD and one of the best-selling anti-AD drugs in the world. Yes, zebrafish has the highest non-trivial harm effect level (NOAEL) among the six drugs, and it has been found to recover extremely quickly (dyskinesia recovery rate = 53.4-64.0%, P\u003c0). .005; responsiveness = 86.6-175.1%, P\u003c0.0001). Memantin-N-methyl-D-asparagine receptor antagonist

　　In addition to acetylcholinesterase inhibitors, Memantin is another important anti-AD drug approved for sale. -Methyl-D-aspartate receptor antagonist. Memantin can block N-methyl-D-aspartate receptors, reduce calcium overflow to the death of nerve cells, and reduce AD symptoms. After treatment with low, medium and high concentrations, the response ability of zebrafish was significantly improved, but the recovery rate of dyskinesias did not change significantly. This result indicates that the recovery rate of motor deficits in this model may not be an indicator of manmantin, but the reactivity is used as a parameter for screening aspartic acid against the anti-AD drug N-methyl D. I can do it. Body drag. Thioflavin T (ThT), flurbiprofen, rosiglitazone Aβ (amyloid β) play an important role in the etiology of AD and increase IDE (insulin degrading enzyme) levels to activate PPARγ receptors and promote Aβ in the brain break down. ThT is an Aβ aggregation inhibitor, but fulviprofen and rosiglitazone (which have been approved as non-steroidal anti-inflammatory drugs and hypoglycemic agents) are used as gamma secretase inhibitors and rosiglitazone, respectively, in clinical trials PPARγ agonist.

　　The recovery rate of dyskinesia cannot be assessed. Ossiglitazone can be explained by the mechanism of Losiglitazone. Rosiglitazone plays a role in the Aβ degradation step after the aggregation and formation steps. Therefore, rosiglitazone may take longer to affect the recovery rate of movement disorders. It’s worth noting that all

　　All zebrafish treated with 10 μM flurbiprofen died. These results indicate that high concentrations of gamma secretase inhibitors and aluminum ions may have toxic side effects on zebrafish. Another reason may be due to the side effects of gamma secretase inhibitors, which may affect the Notch signaling pathway in nerve cell growth. The results are consistent with the mouse model, which can be used to screen gamma secretase inhibitors and anti-AD drugs.

　　AM-117-acetylcholinesterase inhibitor and metalophilic chelator

　　Research shows that the metal ion level of AD patients is 3-7 times that of healthy people and brain metals (iron, copper, zinc). ) But lower levels may help treat AD. In addition, inhibiting acetylcholinesterase and reducing metal ions may have a synergistic effect on AD treatment. In addition, these two drugs can also reduce Aβ aggregation through different mechanisms. Therefore, our laboratory designed AM-117 (an acetylcholinesterase inhibitor and a metal chelator) as MTDL (multi-target targeting ligand) (no published data). This is not surprising. Our data indicate that the measurement of the recovery rate from dyskinesia is a dose-dependent increase in AM-117 treatment.