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Method for detecting activity of anti-angiogenesis protein factor or pro-angiogenesis protein factor via zebra fish embryo model and application method thereof

A technology for zebrafish embryos and new blood vessels, which is applied in biological testing, pharmaceutical formulations, material inspection products, etc., can solve the problems of complex test methods, unfavorable drug screening, long test cycle, etc., and achieve simple drug administration and fast reproduction speed , The effect of simple detection method

Inactive Publication Date: 2006-11-22
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current methods used to detect angiogenesis are too complicated to carry out drug screening. The endothelial cell proliferation and migration test is not only a complex test method, but also an in vitro test; chicken embryo chorioallantoic membrane neovascularization test, rabbit corneal neovascularization test Generation tests and mouse tumor models, etc., are in vivo tests, but the test cycle is long and the method is cumbersome

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Derived from the detection of anti-angiogenesis activity of blue shark cartilage protein component 1

[0033] 1. Embryo collection Take 15 pairs of wild zebrafish in the morning, freely combine and divide them into 5 groups to mate and hatch embryos, collect 1200 zebrafish embryos, suck out the sediment, put them in the hatching solution (distilled water), and store them in a 27°C incubator for 22 hours. Take out the white embryos (dead) in time to prevent the water quality from deteriorating; during this period, the embryos can obtain nutrients through the yolk and do not need other nutrients.

[0034] 2. Preparation of protein component 1. Shark cartilage protein component 1 to be tested was separated and purified by guanidine hydrochloride extraction, fractional precipitation with acetone, ultrafiltration, gel filtration chromatography and other techniques using blue shark cartilage as raw material.

[0035] 3. Add protein component 1. After the zebrafish e...

Embodiment 2

[0046] Example 2 Detection of anti-angiogenic activity of protein component 2 derived from blue shark cartilage

[0047] 1. Embryo collection Take 12 pairs of wild zebrafish in the morning, freely combine and divide them into 4 groups to mate and hatch embryos, collect 1000 zebrafish embryos, suck out the sediment, put them in the hatching solution, and store them in the incubator at 29°C for 22 hours. Use scientific standards to identify dead and alive at any time; dead embryos are white and should be taken out in time to prevent water quality from deteriorating.

[0048] 2. Preparation of protein component 2. Using blue shark cartilage as raw material, pure protein component 2 was obtained by separation and purification through guanidine hydrochloride extraction, fractional precipitation with acetone, ultrafiltration and gel filtration chromatography, with a molecular weight of 1550 Dales pause.

[0049] 3. Add protein component 2. After the zebrafish embryo develops for 22...

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Abstract

The invention relates to the activity detection method for the new-type anti or promote angiogenic factor by zebra fish embryo model. Wherein, after hatching, adding former protein constituent to culture for some time at constant temperature; observing its development; then, dyeing blood vessel, observing with cubic microscope, sampling and storing image. This invention is simple and fit to wide application with low cost.

Description

technical field [0001] The invention relates to a method for detecting the activity of an anti-angiogenic protein factor or a pro-angiogenic protein factor using a zebrafish embryo model and its application, especially a method for detecting the activity of an anti-angiogenic protein factor and its application. Background technique [0002] Malignant tumor is a major disease that endangers human health, so it is also a hot spot in current global research. Currently commonly used antineoplastic drugs have strong toxic and side effects because of their poor specificity, and an ideal drug has not been obtained so far. Anti-tumor drugs targeting angiogenesis have good specificity and have received extensive attention in recent years. It has been found that inhibiting the process of angiogenesis can effectively inhibit tumor growth. Many angiogenesis inhibitors have been shown to be effective in various animal models. Exhibits strong antitumor activity. Compared with commonly u...

Claims

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Application Information

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IPC IPC(8): G01N33/68A61K49/00
Inventor 林秀坤郑兰红杨少丽牛荣丽胡朝欣
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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