Application of exogenous recombinant protein reelin in the preparation of drugs for the treatment of cerebral hemorrhage

A recombinant protein, exogenous technology, applied in the field of biomedicine, can solve problems such as unreported effects, and achieve the effect of reducing pyroptosis

Active Publication Date: 2022-05-24
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of Reelin as an exogenous drug in cerebral hemorrhage has not been reported so far

Method used

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  • Application of exogenous recombinant protein reelin in the preparation of drugs for the treatment of cerebral hemorrhage
  • Application of exogenous recombinant protein reelin in the preparation of drugs for the treatment of cerebral hemorrhage
  • Application of exogenous recombinant protein reelin in the preparation of drugs for the treatment of cerebral hemorrhage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Establishment of intracerebral hemorrhage model

[0023] (1) 24 ICR mice weighing 20-25g were randomly divided into 8 groups (n=3 / group), namely: simple control group (Sham group), 1h, 6h, 12h, 1d, 2d, 3d, 7d group. The modeling steps were as follows: Mice were anesthetized with 4% chloral hydrate by intraperitoneal injection. Depth of anesthesia was observed in mice by eyelid response, skin pinch, or toe reflex. Subsequently, the hair of the mouse head was removed, sterilized with 75% alcohol for several times, the scalp was lifted and cut about 2 cm along the midline, the periosteum was peeled off, and the head of the mouse was fixed with a stereotaxic instrument. Select 1.0mm anterior to the bregma and 2.0mm to the left of the midline, drill the skull with a ball drill to the meninges, pierce the microsyringe vertically 3.5mm, inject 0.5μl of collagenase IV at a constant speed within 5min, and keep the needle for 5min, then withdraw the needle slowly. T...

Embodiment 2

[0026] Example 2: Temporal changes in the expression of pyroptosis-related proteins in different brain regions after intracerebral hemorrhage

[0027] The mice were anesthetized at different time points (1h, 6h, 12h, 1d, 2d, 3d, and 7d) after the intracerebral hemorrhage model was established in the Sham group and 1d after the establishment of the intracerebral hemorrhage model, and the injured side striatum and Cortex, the separated and extracted brain tissue was added to the cell lysate for low temperature homogenization and lysis by ultrasonic cell disruptor, and the supernatant was extracted after low temperature centrifugation for 20 min, and the protein concentration was measured. The proteins were separated by gel electrophoresis. Then, the proteins on the separation gel were transferred to PVDF membranes for 60 min. The PVDF membranes with the proteins transferred were blocked with BSA for 2 h and incubated in BSA-diluted antibodies for 12-14 h at 4°C, followed by PBST....

Embodiment 3

[0029] Example 3: Occurrence and distribution of pyroptosis in different cell types after intracerebral hemorrhage

[0030] Double immunofluorescence staining was performed on the striatum and cortex on the injured side on the 1st day after intracerebral hemorrhage to observe and analyze the relationship between the pyroptosis-related protein ASC and neurons, microglia and astrocytes. The mice were anesthetized 1 day after the establishment of the mouse cerebral hemorrhage model, and the brain tissue was taken out by internal fixation in 4% paraformaldehyde, and then externally fixed for 1 day, followed by sucrose gradient dehydration (20%, 30%, 40%). After the brain tissue sank to the bottom in the sucrose solution, the coronal frozen section of the brain tissue was started. The frozen sections were placed at room temperature for 30 minutes, then fixed in 4% paraformaldehyde for 30 minutes, and washed with PBST solution for several times, five minutes each time. After that, ...

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Abstract

The invention discloses the application of an exogenous recombinant protein Reelin in the preparation of a drug for treating cerebral hemorrhage, belonging to the technical field of biomedicine. The invention discloses the time course change process of pyroptosis in striatum and cortex tissue after cerebral hemorrhage; the distribution of pyroptosis in different cell types in striatum and cortex tissue after cerebral hemorrhage; proves that Reelin improves striatum after cerebral hemorrhage It is proved that Reelin can reduce the pyroptosis of striatum and cortical neurons after cerebral hemorrhage; the present invention provides a basis for using Reelin as a new type of drug for the treatment of cerebral hemorrhage.

Description

technical field [0001] The invention relates to the application of an exogenous recombinant protein Reelin in the preparation of a medicament for treating cerebral hemorrhage, and belongs to the technical field of biomedicine. Background technique [0002] Intracerebral hemorrhage refers to hemorrhage caused by rupture of blood vessels in the non-traumatic brain parenchyma. Although the incidence of cerebral hemorrhage is relatively low in stroke rates, cerebral hemorrhage can lead to extremely high mortality and disability. As a serious public health problem, little progress has been made in clinical treatment, and no drug treatment has been clinically approved to effectively increase survival or improve the quality of life of survivors. And the fatality rate is still at a high level. Therefore, it is imperative to fully understand the pathological changes of cerebral hemorrhage and seek effective treatment methods. [0003] Pyroptosis, also known as cell inflammatory ne...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P7/02A61P9/10
CPCA61K38/1709A61P7/02A61P9/10
Inventor 张明阳单海燕张佳欣杨春
Owner SUZHOU UNIV
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