Small peptide with nerve protection effect and application
A technology of neuroprotection and action, applied in the field of biomedicine, can solve the problems of adverse reactions of antiepileptic drugs, interference and combination, etc., and achieve the effect of improving the survival of neurons
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Embodiment 1
[0018] Example 1 Design of Small Peptides with Neuroprotective Effects
[0019] A small peptide was designed according to the highly conserved sequence of the LBD domain of the GluK2 protein and the CUB binding region (ERMESPID) of NETO2, and a small shuttle peptide (YGRKKRRQRRR) was added to its front end to allow it to enter the cell and interfere with the binding of Gluk2 and NETO2. The amino acid sequence of the small peptide with neuroprotective effect is: YGRKKRRQRRRERMESPID. Small peptides were synthesized by Shanghai Qiangyao Biotechnology Co., Ltd.
Embodiment 2
[0020] Example 2 The protective effect of small peptides on neuron damage after KA-induced epilepsy
[0021] (1) Establishment of rat epilepsy model
[0022] The epilepsy model was induced by KA (0.6 μg / 10 μl) in the lateral ventricle (the control group was given the same dose of solvent), SD rats (230-250 g) were anesthetized with isoflurane by means of an animal anesthesia machine, and fixed in a mouse brain stereotaxic instrument. The skull remains in a horizontal position. Disinfect the skin, incise the scalp to expose the skull, and wash with H 2 o 2 Caulter exposes bregma. Locate the lateral ventricle: 0.8mm posterior to the anterior bregma, 1.5mm lateral to the right, and 3.5mm downward from the surface of the skull. Drill a hole with a 1.5mm drill bit, and withdraw the cerebrospinal fluid with a microsyringe to verify the correctness of the position. The establishment standard of KA-induced epilepsy model is according to Racine classification: grade 1 is closed ey...
Embodiment 3
[0025] Example 3 Effects of small peptides on the interaction of GluK2-NETO2 after KA-induced epilepsy
[0026] (1) Sample preparation
[0027] SD male rats were randomly divided into sham operation group sham and KA administration 6h after modeling, small peptide and control peptide (sham, KA6h, Tat-GluK2+KA, Tat-scramble-GluK2+KA). Then the brain was quickly decapitated, placed in ice-cold PBS, and the CA1 and CA3 / DG regions of the hippocampus were separated, and frozen in liquid nitrogen for future use. Homogenize to extract cytoplasmic protein, and the operation is carried out in an ice bath: take out the hippocampus in liquid nitrogen, add ice-cold homogenization buffer at a ratio of 1:10 (w / v), and homogenize at a high speed with a Glas-Col homogenizer, each The time and frequency of sample homogenization were fixed, and then the supernatant was transferred with an Eppendorf-5840R desktop centrifuge at 14,000rpm, 4°C, for 30min, which was mainly cytoplasmic protein. Th...
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