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Nerve injury repair promoting active polypeptide HW-P3 sourced from sun frogs and application of active polypeptide HW-P3

A HW-P3, nerve injury technology, applied in the field of biomedicine, can solve the problems of increasing morbidity and mortality, allergic reactions, inflammatory reactions and aggravated hemorrhagic transformation, etc., and achieves excellent neuroprotective activity, easy synthesis, and low cost Effect

Active Publication Date: 2022-08-09
KUNMING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Severe allergic reactions may occur after the use of rtPA, and reperfusion after thrombolysis will cause serious secondary damage to brain tissue and cerebrovascular, aggravate the inflammatory response and the risk of hemorrhagic transformation (HT), increasing the disability rate and mortality

Method used

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  • Nerve injury repair promoting active polypeptide HW-P3 sourced from sun frogs and application of active polypeptide HW-P3
  • Nerve injury repair promoting active polypeptide HW-P3 sourced from sun frogs and application of active polypeptide HW-P3
  • Nerve injury repair promoting active polypeptide HW-P3 sourced from sun frogs and application of active polypeptide HW-P3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Screening of the active polypeptide HW-P3 for promoting nerve injury repair

[0018] Step 1: The sample comes from the skin of the frog. Adult zebra frogs (from Yunnan Province, China) were housed in a 21 cm × 21 cm × 15 cm rearing tank for 7 days for acclimatization, with regular water changes and mealworms as feed. After rinsing the frog's skin with deionized water, the frogs were quickly killed and their skin was harvested by disrupting the brain.

[0019] Step 2: Construction of the frog cDNA library: In order to screen the cDNA encoding mature HW-P3, the polymerase chain reaction (PCR) template was obtained from the cDNA synthesized by SMART technology and the 5' PCR primer (5' PCR) synthesized by BGI Company (China). '-CCAAA (G / C) ATGTTCACC (T / A) TGAAGAA-3') and 3' PCR primer (5' ATTCAGGCCGAGGCC GACA TG-3'). The obtained PCR product was cloned into E. coli DH5a-active cells and DNA sequenced using an Applied Biosystems DNA Sequencer (ABI 3730XL, Foster...

Embodiment 2

[0021] Example 2 Detection of the activity of polypeptide HW-P3 in promoting nerve damage repair

[0022] 1. In vitro activity identification

[0023] Experimental method: In vitro cellular oxygen-glucose deprivation / reoxygenation experiment

[0024] PC12 cells were cultured in DMEM / F12 medium containing 10% fetal bovine serum and 1% antibiotics (100 U / mL penicillin, 100 U / mL streptomycin) at 37°C and 5% CO 2 . PC12 cells were cultured in 96-well plates (5000 cells / well). After the cells adhered, the cells in the negative control group were cultured in the same manner as described above. The PC12 cells except the negative control group were washed twice with PBS, replaced with sugar-free DMEM, and placed in 95% N. 2 and 5%CO 2 cultivated in an environment. After 4 hours of reoxygenation, the glucose-free DMEM medium of PC12 cells after the establishment of the oxygen-glucose deprivation / reoxygenation model was replaced with the original complete medium, and placed at 37°C...

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Abstract

The invention discloses a nervous injury repair promoting active polypeptide HW-P3 derived from a sun frog and application of the nervous injury repair promoting active polypeptide HW-P3. And the amino acid sequence of the polypeptide HW-P3 is FLPAAICLVIKTC. The application refers to the application in preparation of drugs for treating or preventing brain nerve injury. The invention provides a novel polypeptide HW-P3 derived from the skin of a sun frog. The polypeptide HW-P3 has the activity of promoting nerve injury repair in vivo and in vitro. And the HW-P3 only has 13 amino acids, is easy to synthesize and low in cost, provides a new approach for development of nerve injury repair drugs, and also provides a new direction for development of cerebral arterial thrombosis treatment drugs.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an active polypeptide HW-P3 derived from frogs for promoting nerve damage repair and application thereof. Background technique [0002] Ischemic stroke is a general term for brain tissue necrosis caused by stenosis or occlusion of the blood supplying arteries to the brain and insufficient cerebral blood supply and oxygen supply, accounting for about 70-80% of all strokes. Cerebral ischemia and hypoxia lead to brain tissue necrosis, hemianopia, aphasia, sensory disturbance, and disturbance of consciousness. [0003] The high disability and high mortality of ischemic stroke has caused a serious burden on the world and families, but the effective treatment methods and drugs are very limited. At present, recombinant tissue plasminogen activator (rtPA) thrombolytic therapy is recognized as the most effective treatment for stroke, but it has a strict time window (4.5 h...

Claims

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Application Information

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IPC IPC(8): C07K14/46A61K38/17A61P25/00A61P9/10
CPCC07K14/463A61P9/10A61P25/00A61K38/00Y02A50/30
Inventor 杨新旺王滢李依林尹赛格王颖蕾杨美凤刘承杏冯成安刘乃心吴雨桐
Owner KUNMING MEDICAL UNIVERSITY
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