Application of manganese type high-stability superoxide dismutase in prevention or treatment of cerebral apoplexy

A high stability, superoxide technology, applied in the field of biomedicine, can solve problems such as side effects

Inactive Publication Date: 2021-06-04
CARRY HEALTH BIOPHARMACEUTICALS HANGZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These drugs are mainly small molecular substances, which are used for free radical scavenging and post-injury protection. They have a certain effect on the treatment of stroke, but they also have side effects

Method used

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  • Application of manganese type high-stability superoxide dismutase in prevention or treatment of cerebral apoplexy
  • Application of manganese type high-stability superoxide dismutase in prevention or treatment of cerebral apoplexy
  • Application of manganese type high-stability superoxide dismutase in prevention or treatment of cerebral apoplexy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the preparation of manganese type high stability superoxide dismutase (MS-SOD)

[0028] Using the genome of thermophilic bacteria HB27 (purchased from the ATCC cell bank in the United States, the deposit number is ATCC BAA-163) as a template, the target gene was amplified with the following primer sequence: Forward primer: 5'-aagaattcatgccgtacccgttcaagct-3' (SEQ ID NO.1) Reverse primer: 5'-ctgtcgactcaggctttgttgaagaac-3' (SEQ ID NO.2); the amplified product was recovered with a recovery kit (purchased from Sangon Bioengineering Shanghai (Share) Co., Ltd.), and the enzymes EcoRI and SalI double enzyme digestion, and connected to the plasmid vector pET28a (+) (purchased from Sangon Bioengineering Shanghai (Co., Ltd.)) with the same enzyme double enzyme digestion, the recombinant plasmid was transformed into competent Escherichia coli BL21 (DE3) ( Purchased from Sangon Bioengineering (Shanghai (Stock) Co., Ltd.), the strain with the correct MS-SOD nucleotide s...

Embodiment 2

[0032] Embodiment 2: mouse model experiment

[0033] 1. Animal origin

[0034] Male C57BL / 6J wild-type mice were purchased from Guangdong Experimental Animal Center. When implementing the middle cerebral artery occlusion (MCAO) model, male C57BL / 6J wild-type mice were 8 weeks old.

[0035] 2. Grouping and handling of mice

[0036] According to different MS-SOD intervention methods, the mice were divided into:

[0037] (1) Control group (10 mice): 8-week-old healthy mice.

[0038] (2) Model group (10 rats): middle cerebral artery embolism ischemia model group without MS-SOD intervention;

[0039] The modeling process is shown in part 3 below;

[0040] (3) MS-SOD1 group (10 rats): 4-week-old mice drank drinking water added with MS-SOD for a total of 4

[0041]Week (3U / day / gram body weight), and then implement middle cerebral artery embolism ischemia modeling;

[0042] (4) MS-SOD2 group (10 rats): After the middle cerebral artery embolism ischemia model was established, af...

Embodiment 3

[0057] Example 3: MS-SOD treatment of zebrafish ischemic stroke model experiment

[0058] 1. Animal origin

[0059] Zebrafish were provided by the Animal Center of Hangzhou Huante Biotechnology Co., Ltd.

[0060] 2. Grouping of animals

[0061] Experimental group 1 Normal control group

[0062] Experimental group 2 Model control group

[0063] Experiment 3 group positive control drug aspirin 50μg / mL

[0064] Experiment 4 group MS-SOD 222μg / mL

[0065] Experimental group 5 MS-SOD 667μg / mL

[0066] Experiment 6 group MS-SOD 2000μg / mL

[0067] 3. Model making

[0068] Ponatinib was administered in a water-soluble manner (1 μg / mL) to treat normal 2 days after fertilization (2dpf) melanin allele mutant Albino strain zebrafish for 18 hours to establish a zebrafish cerebral ischemia model.

[0069] 4. Experimental method

[0070] A total of 180 zebrafish of the Albino strain with melanin allele mutation were randomly selected at 2 days after fertilization (2dpf), and 30 zebr...

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Abstract

The invention relates to application of manganese type high-stability superoxide dismutase in prevention and treatment of cerebral apoplexy. The amino acid sequence of the manganese type high-stability superoxide dismutase is shown as SEQ ID NO: 4. The manganese-type high-stability superoxide dismutase can be used for remarkably preventing and reducing brain injury after cerebral apoplexy, and has a very good prevention and treatment effect on cerebral apoplexy.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the prevention or treatment of stroke. Background technique [0002] Stroke, including ischemic stroke and hemorrhagic stroke, is a type of brain tissue damage caused by blockage of blood vessels or blood vessel rupture and bleeding that prevents blood from flowing into the brain normally. Ischemic stroke accounts for 70% of strokes, and stenosis and occlusion of carotid and vertebral arteries can cause ischemic stroke. After a stroke occurs, it will cause damage to brain tissue, and severe cases are life-threatening. During ischemia and reperfusion, mitochondrial damage, calcium overload, accumulation of free radicals, and inflammatory response may occur. The excessive free radicals produced in the brain lead to an imbalance in the oxidation-antioxidation system, which is an important aspect. Free radicals cause peroxidation of lipids, proteins and nucleic acids, leadin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/44A61P9/10
CPCA61K38/446C12Y115/01001A61P9/10A61K38/44A61K38/00A61P9/00C12N9/0089
Inventor 孟凡国
Owner CARRY HEALTH BIOPHARMACEUTICALS HANGZHOU CO LTD
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