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Use of fbp aldolase in the preparation of drugs for activating AMPK
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A drug and application technology, applied in the direction of drug combination, application, microorganisms, etc., can solve the problems of inhibiting aldolase and the lack of universality of aldolase inhibition, and achieve the effect of overcoming difficulties
Active Publication Date: 2021-09-24
XIAMEN VIVOHEALTHS TECH CO LTD
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In addition, the inventor's preliminary experimental results show that TDZD-8 cannot inhibit aldolase in MEF cells, which at least shows that the inhibition of aldolase by this inhibitor is not universal
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Embodiment 1
[0099] Embodiment 1: shRNA suppresses the expression of ALDOA-C gene
[0100] 1. Experimental materials and main reagents
[0101] Cell line: mouse fibroblast MEF, which is an immortalized MEF, that is, after the fibroblasts are isolated from mouse embryos, they are transfected with SV40 T antigen to make the cells immortal. For the construction method, please refer to the literature Lei Y, Methods MolBiol. 2013; 1031:59-64. Generation and culture of mouse embryonic fibroblasts.
[0102] Human embryonic kidney cells HEK293T (cat.CRL-3216) were purchased from ATCC.
[0103] Vector pLVX-IRES (cat. #631849) was purchased from Clontech.
[0145] Rabbit 548anti-phospho-AMPKα-T172 (cat#2535), purchased from Cell Signaling Technology;
[0146] anti-AMPKα (cat#2532, 1:1000 for IB), purchased from Cell Signaling Technology;
[0147] anti-phospho-ACC-Ser79 (cat.#3661, 1:1000 for IB), purchased from Cell Signaling Technology;
[0148] anti-ACC (cat.#3662, 1:1000 for IB), purchased from Cell Signaling Technology.
[0149] Other primary and secondary antibodies used are as described in Example 1.
[0150] DMEM without glucose (Gibco, cat. 11966) was purchased from Thermofisher.
[0151] 2. Experimental method
[0152] (1) First, with reference to the method described in Example 1, prepare the following figure 1 ALDOA-C knockdown MEF cells are shown. Among them, the shRNA targeting GFP (Green Fluorescent ProteinGreen Fluorescent Protein) is used as the co...
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Abstract
The invention belongs to the field of biomedicine, and relates to the application of FBP aldolase in the preparation of drugs for activating AMPK. The present invention also relates to the preparation of drugs for inhibiting cholesterol synthesis, drugs for reducing fatty acid synthesis, drugs for preventing and / or treating diabetes, drugs for preventing and / or treating tumors, drugs for preventing and / or treating Parkinson's disease, drugs for preventing and / or treating Or use in a drug for the treatment of Alzheimer's disease or a drug for prolonging the lifespan of an organism. The invention realizes that FBP aldolase can be used as a target to develop drugs for activating AMPK, which overcomes the difficulties in the prior art of directly using AMPK as a drug target, and has good application prospects.
Description
technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of FBP aldolase in the preparation of drugs for activating AMPK. The present invention also relates to the preparation of drugs for inhibiting cholesterol synthesis, drugs for reducing fatty acid synthesis, drugs for preventing and / or treating diabetes, drugs for preventing and / or treating tumors, drugs for preventing and / or treating Parkinson's disease, drugs for preventing and / or treating Or use in medicines for treating Alzheimer's disease or medicines for prolonging the lifespan of mammals. Background technique [0002] 5'-adenosine monophosphate-activated proteinkinase (AMPK) is an important molecule that regulates the energy balance of cells and the body. AMPK consists of three different subunits, each of which has several isoforms: α subunit (α1 or α2); β subunit (β1 or β2); and γ subunit (γ1, γ2 or γ3 ); there are a total of 12 possible isoforms of heterotrimer...
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