Application of human RTN4B protein in preparation of medicine for healing wound

A wound healing and drug technology, applied in the fields of genetic engineering and medicine, which can solve the problem of not finding the variant splicing book, etc.

Inactive Publication Date: 2007-08-15
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no variant spliced ​​version of RTN3 has been found so far

Method used

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  • Application of human RTN4B protein in preparation of medicine for healing wound
  • Application of human RTN4B protein in preparation of medicine for healing wound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Chicken embryo allantoic membrane (CAM) test

[0051] Material:

[0052] 1. Hailan white-shell eggs were purchased from Shanghai Poultry Breeding Co., Ltd.

[0053] 2. Elbow tweezers, scissors.

[0054] 3. Alcohol noodles.

[0055] 4. Hole punch.

[0056] 5. Petri dish.

[0057] 6. Dexamethasone Sodium Phosphate Injection.

[0058]7. Whatman type I filter paper.

[0059] 8. bFGF.

[0060] step:

[0061] 1. Prepare filter paper: prepare a suitable amount of filter paper (5 mm in diameter) with a puncher sterilized by 70% ethanol, put it in a petri dish for autoclaving and drying. Add 8 ul / piece of dexamethasone sodium phosphate injection to each piece of filter paper on the ultra-clean workbench, and air-dry for 30 minutes under sterile laminar flow.

[0062] 2. At 37.5 degrees Celsius and 51% relative humidity, cultivate Hy-Line white shell eggs. On the third day of embryonic development, open a window and put filter paper in it. The specific operat...

Embodiment 2

[0066] Embodiment 2 cornea experiment

[0067] Material:

[0068] KM mice, Shanghai Slack Experimental Animal Co., Ltd.

[0069] Hydron, the full name is Poly (2-hydroxypropyl methacrylate), sigma company

[0070] step:

[0071] 1. Prepare the hydron, order egg white

[0072] The full name of hydron is Poly (2-hydroxypropyl methacrylate), purchased from sigma company. Hydron, also known as hydrogel (hydrogel), is formed by hydration to form a slow-release gel, thereby controlling the release rate.

[0073] Dissolve 32 mg of sucralfate in 72 μl of PBS and mix well, and the hydron is prepared into a 12% solution with absolute ethanol. Mix equal volumes of 50 μl each of the two solutions.

[0074] On the ultra-clean workbench, spot the mixed solution on the parafilm membrane, about 0.5mm in diameter, and each pellet contains about 0.5μl mixed solution, as protruding as possible. After pointing, blow it under UV light for 20 minutes.

[0075] Spot the protein on the pellet...

Embodiment 3

[0087] Example 3 Cell adhesion experiment

[0088] 1. The rtn4b protein was serially diluted with PBS, and the final concentrations were 60 and 600 nM, respectively, and a negative control (pbs and bsa) and a positive control af (Casade biologicals, 5006-100) were set up. Take one 96-well plate, add 50 microliters of each of the above solutions into different wells, and each group has five duplicate wells. Then place it at 4°C overnight.

[0089] 2. Absorb the coating solution the next day, wash each well with PBS, wash twice, add 150 microliters of BSA (bovine serum albumin) with a concentration of 1 mg per milliliter to each well, and place it at 37 degrees to block for 1 hour .

[0090] 3. During the blocking process, digest the cells ECV304 to single cells, count the number of cells with a hemocytometer, and calculate the cell concentration.

[0091] 4. Aspirate the required cell solution, centrifuge at 1000 rpm for 5 minutes, remove the supernatant, add the required vo...

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Abstract

This invention relates to the field of gene engineering and medicament, specifically relates to the application of RTN 4B in preparing wound healing drugs. Healing of wound is a very complicated process. Wound not only changes part of organism, but also initiate systemic reaction to different extent, meanwhile, wound part will bring pain to patients. The inventive RTN 4B can promote angiogenesis, enhance adhesion and migration of blood vessel cell; and can be used for preparing medicament for promoting healing of wound to shorten period of treatment and relieve pain.

Description

technical field [0001] The invention relates to the fields of genetic engineering and medicine, in particular, the invention relates to the application of human protein RTN4B in the preparation of wound healing medicine. Background technique [0002] The RTN family is a new gene family discovered in recent years. Among them, the neuroendocrine-specific protein NSP (neuroendocrine-specific protein) was uniformly named RTN1, which is the first member of this family. According to the comparison of NSP genome sequence, cDNA sequence and nucleic acid sequence database, three other members of the human RTN family (NSP gene I, II, III) were found. Two of them (NSP-like gene I, II) were cloned successively and named RTN2 and RTN3. RTN2 also has three isoforms with the same C-terminus. However, no variant spliced ​​version of RTN3 has been found so far. Notably, two independent transmembrane domains were also found at the C-termini of RTN2 and RTN3, which are thought to be associ...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P17/02
Inventor 余龙季国庆刘星吴燕华唐丽莎
Owner FUDAN UNIV
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