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Chinese hard tick neuropeptide and its preparing method and use

A technology of Ixodes sinensis and neuropeptides, applied in the field of biomedicine, can solve the problems of unclear protein components and sequences, and achieve strong analgesic activity

Inactive Publication Date: 2005-10-19
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

At present, a large number of biologically active molecules with physiological and pharmacological activities such as prostaglandins, anticoagulant factors, immunosuppressive substances and anti-inflammatory proteins have been found in the salivary glands of ticks to overcome the host's defense response. The latest research results show that, The salivary glands of ticks also have proteins with anti-complement activity, which inhibit the host's immune response by inhibiting the activation of complement, but their protein components and sequences are still unclear

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  • Chinese hard tick neuropeptide and its preparing method and use
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  • Chinese hard tick neuropeptide and its preparing method and use

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Embodiment Construction

[0024] 1. Preparation of Ixodes sinensis neuropeptide:

[0025] The hungry male and female Ixodes sinensis adults were glued to the bottom of the watch glass and placed on ice for 20 minutes, and then the Ixodes were cut along the dorsal edge to remove the ganglion. The ganglia were transferred to 0.1M acetic acid, rinsed three times, and finally placed in 0.1M acetic acid and stored at -20°C.

[0026] The ganglion isolated from 10,000 Ixodes sinensis was homogenized in 3ml of 0.1M acetic acid containing 30μl protease mixed inhibitor (Sigma). The homogenate was centrifuged at 5000 rpm for 5min, and the supernatant was collected and washed with a 3kDa membrane (Millipore) filter. The filtrate was passed through 5mm×300mm hypersil ODS2 5μm RP-HPLC (Dalian Yilite). The procedure for passing the column is as follows: first equilibrate the column with sterile water containing 0.1% trifluoroacetic acid for 6 minutes, the flow rate is 0.7ml / min, and load 200 μl each time, and then ...

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Abstract

The present invention belongs to the field of biomedicine technology. Chinese hard tick neuropeptide is a kind of single-stranded small peptide obtained from the ganglion of Chinese hard tick and with molecular weight of 1005.22 dalton, isoelectric point of 8.59, and the first order whole sequence structure of NH2-LVVYPWTK-COOH. It is prepared through adhering Chinese hard tick to the bottom of watch glass and on ice for 20 min, cutting along back side to eliminate ganglion, homogenizing, centrifuging and eliminate precipitate, filtering with membrane, and twice reverse high efficiency liquid phase chromatographic separation and purification. The Chinese hard tick neuropeptide has powerful analgetic activity, and may be used as analgetic medicine.

Description

Technical field: [0001] The invention provides an Ixodes sinensis neuropeptide and its preparation method and application, which belong to the technical field of biomedicine. Background technique: [0002] Ticks are the most harmful blood-sucking parasites to livestock, and to humans, they are the second largest source of disease after mosquitoes. Ticks are widely distributed all over the world, and there are more than 800 species, and there are more than 110 species in my country alone. It can be divided into three categories: hard ticks, soft ticks and narcotic ticks. Ticks are important carriers of many pathogens such as bacteria, fungi, viruses, spirochetes, rickettsiae and protozoa. Ticks can parasitize amphibians, reptiles, birds and mammals, and often change hosts, so they can also transmit many zoonotic diseases to humans, which has brought great harm to human public health and safety, and Lyme disease transmitted by ticks (also known as "Second AIDS"), Q fever an...

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Application Information

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IPC IPC(8): A61K38/08A61P25/04C07K7/06
Inventor 赖仞李健许徐学清杨海龙
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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