Composition and preparation method of SeZn7MT3 and application thereof in preventing and treating Alzheimer's disease
A technology of metallothionein and selenogenesis, applied in the field of biomedicine, can solve problems such as synaptic function damage and destruction of synapse-related protein functions
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Embodiment 1
[0015] Example 1: SeZn 7 Preparation of MT3
[0016] All biochemical reagents and kits were purchased from Sigama or Invitrogen unless otherwise specified.
[0017] (1) Smt-MT3 plasmid construction
[0018] Human metallothionein MT3 and fusion tag protein gene Smt3 were purchased from Guangzhou Funeng Gene Company. The gene carrier plasmid MBPHT-mCherry-2 was purchased from Invitrogen, and the gene primer synthesis was completed by Shanghai Jereh Biotechnology Co., Ltd., using the designed primers P1, P2 (to amplify the Smt3 gene) and P3, P4 (to amplify the MT3 gene) to amplify out the PCR product. The amplified products were detected and verified by 1% agarose gel, and the correct fragments were recovered from the gel. Product 1 and product 2 were mixed at the same concentration ratio as a template, and the Smt3-MT3 sequence was amplified using primers P1 and P4, with a BamH I restriction site GGATCC at its 5' end and a Hind III restriction site at its 3' end Site AAGCTT...
Embodiment 2
[0035] Embodiment 2: Alzheimer's disease mouse model and SeZn 7 Drug efficacy test of MT3
[0036] C57 ordinary mice, male, weighing 24-26 g, 5 months old, SPF grade, were purchased from Shanghai Slack Experimental Animal Co., Ltd. [Experimental animal quality certificate number: SCXK (Shanghai) 2007-0005]. APP / PS1 transgenic mice were purchased from Beijing Zhongke Zesheng Biotechnology Co., Ltd., 5 months old, weighing 24-26g. All mice were free to forage and drink, and were kept at room temperature (23±2)°C. Feed and water were all processed by autoclaving, and all experimental feeding processes were SPF grade. The experimental mice were randomly divided into 3 groups, 10 mice in each group. A: normal C57 mice group, B: APP / PS1 transgenic mice, negative control group (normal saline group); C: SeZn7MT3 administration group, APP / PS1 transgenic mice.
[0037] Test drug name: SeZn 7 MT3;
[0038] Solvent: normal saline;
[0039] Preparation method: Prepare the solution w...
Embodiment 3
[0041] Example 3: Using the Morris water maze to verify the cognitive ability of Alzheimer's mice
[0042] The experimental mice were randomly divided into two groups, with 10 mice in each group. A: APP / PS1 transgenic mice; B: SeZn 7 MT3 administration group, APP / PS1 transgenic mice.
[0043]Device: a circular pool with a diameter of 1m, a height of 50cm, and a water depth of 30cm. The bottom of the pool is white, and the water temperature is kept at 23±2°C. Four equidistant points N, E, S, and W are marked on the pool wall as the starting point of the test. The pool has four quadrants, and a platform is placed in the center of the third quadrant (the distance between the platform and the center of the pool wall is equal); it is submerged 1 cm below the water, so that the platform is invisible. There are abundant reference clues around the pool (triangles, squares, circles, and rhombuses of different colors are placed in each quadrant) and remain unchanged for the mice to us...
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