Scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia and preparation method and application of scorpion toxin active polypeptide HsTx1

An active polypeptide, cerebral ischemia technology, applied in the field of biomedicine, to achieve the effect of strong anti-cerebral ischemia and hypoxia, strong improvement of motor dysfunction activity, and strong activity improvement

Active Publication Date: 2019-12-06
KUNMING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, active peptides that improve cereb

Method used

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  • Scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia and preparation method and application of scorpion toxin active polypeptide HsTx1
  • Scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia and preparation method and application of scorpion toxin active polypeptide HsTx1
  • Scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia and preparation method and application of scorpion toxin active polypeptide HsTx1

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preparation example Construction

[0028] The preparation method of the scorpion toxin active polypeptide HsTx1 with improved cerebral ischemia and hypoxia of the present invention comprises the following steps:

[0029] A. Take the hook-shaped stinger secretion on the tail end of the tropical rainforest scorpion by electrical stimulation, dissolve it in PBS, freeze-dry it in vacuum to obtain material a, and store it at -80°C for later use;

[0030] B. Dissolving material a in water to obtain material b, eluting material b with Tris-HCl buffer, collecting the eluent to obtain material c;

[0031] C, subjecting material c to high performance liquid chromatography reversed-phase chromatography to obtain material d;

[0032] D. The material d is subjected to the second high-performance liquid chromatography reversed-phase chromatography to obtain the target substance, the scorpion toxin active polypeptide HsTx1, which can improve cerebral ischemia and hypoxia.

[0033] The water described in the B step is deioniz...

Embodiment 1

[0043] Separation, purification and identification of scorpion toxin active polypeptide HsTx1 that can improve cerebral ischemia and hypoxia

[0044] 1. Separation and purification

[0045] Live tropical rainforest scorpions were collected from Xishuangbanna, Yunnan. The hook-shaped stinger secretions (dissolved in PBS) were obtained by electrical stimulation, vacuum freeze-dried, and stored at -80°C for later use.

[0046] The first step: Sephadex G75 molecular sieve:

[0047] The freeze-dried powder of skin secretions was obtained according to the above method, dissolved in deionized water, and 500 μl (protein content: 100 mg) was pre-equilibrated with 20 mM Tris-HCl buffer (pH=7.8, containing 0.1 M NaCl) for 24 h. Sephadex G75 (GE Healthcare, ultra-fine) column (length 40 cm, inner diameter width 1.5 cm), eluted with the same buffer, the flow rate was 1.5 ml / 10 min, collected once every 10 min, and measured at 280 nm of absorbance.

[0048] The second step: the first hig...

Embodiment 2

[0056] Anti-ischemia-hypoxia activity of scorpion toxin active polypeptide HsTx1 which can improve cerebral ischemia-hypoxia

[0057] The present invention uses the animal model of cerebral ischemia and hypoxia (MCAO) experiment to detect the anti-cerebral ischemia and hypoxia activity of HsTx1, and the results show that the scorpion toxin anti-cerebral ischemia and hypoxia activity polypeptide HsTx1 has a strong activity of improving cerebral ischemia and hypoxia , and can effectively improve the dysfunction after cerebral ischemia and hypoxia.

[0058] SPF grade male SD rats weighing 250-280 g were selected for the experiment, and the rats were randomly divided into three groups, sham operation group, MCAO control group, MCAO+1 nmol / kg group, MCAO+2 nmol / kg group, MCAO+ 5 nmol / kg group, 7 rats in each group, and the rats were trained to cross the balance beam before operation. First, the rats were anesthetized by intraperitoneal injection of 3% pentobarbital sodium (30 mg / k...

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Abstract

The invention discloses scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia and a preparation method and application of the scorpion toxin active polypeptide HsTx1. An amino acid sequence of the scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia is AGKKERAGSRRTKIVMLKCIREHGH. According to the scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia and the preparation method and the application of the scorpion toxin active polypeptide HsTx1, the purified and obtained rainforest scorpion toxin polypeptide HsTx1 shows strong anti-cerebral ischemia and hypoxia activity and activity of improvement of motor dysfunctions in a cerebral ischemia and hypoxia model (MCAO) experiment, and it is indicated that the 'scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia' has a great prospect of medical application; and the scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia has the beneficial characteristics of strong activities of efficiently improving cerebral ischemia and hypoxia and the motor dysfunctions.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia, as well as its preparation method and application. Background technique [0002] As we all know, cerebral ischemia is one of the most common acute cerebrovascular diseases, and has been recognized as the third leading cause of mortality and disability in the world, which has caused huge economic and psychological burdens on individuals and society. Acute cerebrovascular disease is a group of diseases that cause damage to brain tissue due to sudden rupture of blood vessels in the brain or blockage of blood vessels that prevent blood from flowing into the brain, including ischemic and hemorrhagic strokes. Among them, the incidence of ischemic stroke accounts for 60% to 70% of the total number of strokes, and its symptoms include sudden weakness of one side of the head, face ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/14C07K1/20A61K38/17A61P9/10A23L33/18
CPCC07K14/43522A61P9/10A23L33/18A61K38/00A23V2002/00A23V2200/30A23V2250/55
Inventor 杨新旺孙俊杨美凤王滢宋咏丽尹赛格
Owner KUNMING MEDICAL UNIVERSITY
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