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Application of human FGF21 (fibroblast growth factor 21) in preparing medicines for treating stroke

A growth factor and fibroblast technology, applied in the field of medicine, can solve the problems of little knowledge, limit the ability of brain nerve repair and reconstruction, and aggravate functional deficits, and achieve the effect of promoting nerve regeneration, promoting reconstruction, and reducing neuroinflammation.

Active Publication Date: 2017-08-18
杭州生物医药创新研究中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, despite abundant reports of diabetic-induced renal and retinal microvascular complications, the mechanisms of diabetic-induced neuropathy, such as stroke, remain poorly understood
Patients with T2D often have hyperglycemia, dyslipidemia, and insulin resistance, factors that exacerbate long-term functional deficits after stroke and limit the brain's inherent neural repair and rebuilding capacity

Method used

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  • Application of human FGF21 (fibroblast growth factor 21) in preparing medicines for treating stroke
  • Application of human FGF21 (fibroblast growth factor 21) in preparing medicines for treating stroke
  • Application of human FGF21 (fibroblast growth factor 21) in preparing medicines for treating stroke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1 Obtaining exogenous recombinant human fibroblast growth factor 21

Embodiment 11

[0090] The construction of embodiment 1.1 high expression bacterial strain

[0091] Firstly, the corresponding DNA sequence was optimally designed according to the protein sequence of human FGF21 (NP_061986.1) on GenBank, as shown in SEQ ID NO.1. Subsequently, the corresponding DNA fragment was artificially synthesized, which had an NdeI restriction site at the 5' end and a BamHI restriction site at the 3' end. The resulting DNA of FGF21 was double digested with NdeI and BamHI, and the expression empty vector pET3c was treated with the same enzymes. The two digested fragments were ligated with T4 DNA ligase, transformed into competent Escherichia coli DH5α, and positive clones were screened for resistance to Ampicillin, and DNA sequence analysis was performed to ensure that the obtained cDNA sequence of FGF21 was correct. The resulting recombinant plasmid was named pET3c-FGF21. The recombinant plasmid pET3c-FGF21 was transformed into Escherichia coli BL21(DE3), and transform...

Embodiment 13

[0096] Example 1.3 Purification of FGF21

[0097] 1) Preparation of inclusion bodies

[0098] Suspend the wet cells in 10 times volume (w / v) buffer containing 25mmol / L Tris (pH 8.0), 150mmol / L NaCl, 10mmol / L EDTA-2Na; according to the required FGF21 wet cells, Weigh lysozyme and DNase at a ratio of 1:1000 (w / w) and 1:10000 (w / w), dissolve them in the lysate and add them to the bacterial suspension, stir and lyse at room temperature overnight, and microscopically check for cracks. Bacterial solution until there are no complete bacterial cells in the field of view, then use a high-speed refrigerated centrifuge (9000r / min, 4°C) to separate and collect the precipitate.

[0099] Suspend the precipitate from the previous step in 15 times the volume (w / v) of the wet cells containing 25mmol / L Tris (pH8.0), 150mmol / L NaCl, 10mmol / L EDTA-2Na, 0.2% deoxycholic acid In the sodium washing liquid I, fully stir to dissolve and mix well, then use a high-speed refrigerated centrifuge (9000r / ...

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Abstract

The invention discloses application of human FGF21 (fibroblast growth factor 21) in preparing medicines for treating stroke. The medicine for treating the stroke has the advantages that the integrity of blood brain barrier is protected, the neuroinflammation in brain is inhibited, the rebuilding of blood vessels and white matter in the brain is promoted, the restoration of nerve function after stroke is improved, and the brain injury of a stroke patient and a diabetes and stroke patient is greatly improved.

Description

technical field [0001] The invention relates to a new application of human fibroblast growth factor 21 (FGF21), in the technical field of medicine, in particular to the application of human FGF21 in the preparation of medicines for treating cerebral apoplexy. Background technique [0002] Stroke, commonly known as "stroke", has the characteristics of high morbidity, high disability, high recurrence and high mortality. . Stroke is mainly divided into two categories: ischemic stroke and hemorrhagic stroke, of which about 80% are ischemic stroke. Ischemic stroke is caused by a transient or permanent decrease in blood flow in the arteries of the brain. Brain-protective drugs are currently a research hotspot in the neurological community. At present, the only drug that has been proven effective and recommended as a routine treatment is aspirin. The administration of r-tPA within 3 hours after the onset of ischemic stroke and the administration of pre-arterial urokinase within ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/18A61P9/10A61P25/00
CPCA61K38/1825
Inventor 李校堃王晓英林丽王晓杰王怡
Owner 杭州生物医药创新研究中心
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