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Method for purifying skin secretions of tailed amphibian ranodon sibiricus and application

A technology for skin secretions and tailed amphibians, which is applied in the field of biomedicine and can solve problems such as long retention time and difficult to wash off sticky substances

Inactive Publication Date: 2014-05-07
XINJIANG NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] When the Xinjiang northern salamander is artificially raised, whenever the Xinjiang northern salamander is caught by hand, the sticky substance secreted by its skin is difficult to wash off from the hands, and the retention time is longer

Method used

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  • Method for purifying skin secretions of tailed amphibian ranodon sibiricus and application
  • Method for purifying skin secretions of tailed amphibian ranodon sibiricus and application
  • Method for purifying skin secretions of tailed amphibian ranodon sibiricus and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The method for purifying the skin secretions of the tailed amphibian, the Xinjiang northern salamander, is as follows:

[0038] 1) The collected solution was frozen in a -80°C ultra-low temperature refrigerator for 24 hours, then lyophilized and vacuum-dried, and concentrated to make a dry powder.

[0039] 2) The obtained dry powder can be stored in a -20°C refrigerator for a long time. Before use, PBS is dissolved and sterilized with a bacterial filter membrane, and then stored in a -20°C refrigerator for later use.

Embodiment 2

[0040] Example 2: The effect of Xinjiang northern salamander skin secretion on tumor cell growth inhibition

[0041] Step 1: Take Hela cells, A549 cells and HepG2 cells respectively and culture them at 37°C and 5% CO 2 In the cell culture incubator, use DMEM high glucose medium containing 10% fetal bovine serum and 1% double antibody. After culturing to a certain number, the tumor cells in the logarithmic growth phase were digested and blown with 0.25% trypsin to make a single-cell suspension, and 5×10 3 The density of cells / well was inoculated in 96-well culture plate and cultured overnight in the incubator.

[0042] Step 2: After the cells adhere to the wall, they are divided into groups according to the addition of different drugs:

[0043] Negative control group: no drug treatment.

[0044] Positive control group: 5-fluorouracil (5-FU) with a concentration of 2.5 mg / ml and 10 gradients of 2-fold dilution followed by drug addition.

[0045] Medication group: the sample ...

Embodiment 3

[0051] Example 3: Morphological changes of tumor cells by the skin secretions of the northern salamander in Xinjiang

[0052] Step 1: Culture A549 cells at 37°C, 5% CO 2 In the cell culture incubator, use DMEM high glucose medium containing 10% fetal bovine serum and 1% double antibody. After being cultivated to a certain number, they are divided into groups according to whether to add drugs:

[0053] Negative control group: no drug treatment

[0054] Sample group: add 1 μg / mL of drug.

[0055] Step 2: 6h, 12h, and 24h after drug treatment, clip the slides of the control group and the experimental group respectively, pay attention to marking the front and back sides, and wash them with 0.1M phosphate buffer (twice).

[0056] Step 3: Use 2.5% glutaraldehyde solution (prepared in PBS) to fix in a fume hood at room temperature for 2 hours or even longer. After fixation, rinse with 0.1M pH7.4 phosphate buffer and distill 3 times, 10min each time.

[0057] Step 4: After washin...

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Abstract

The invention relates to a method for purifying skin secretions of tailed amphibian ranodon sibiricus and an application and relates to the field of biomedicine, wherein the skin secretions are antineoplastic. The method includes using double distilled water to wash the skin secretions of the ranodon sibiricus, collecting the skin secretions, and subjecting the skin secretions to suction filtration to remove impurities; freezing collected liquids of the impurity-removed skin secretions of the ranodon sibiricus and drying and concentrating the collected liquids to obtain skin secretion dry powders of the ranodon sibiricus; and preserving the skin secretion dry powders of the ranodon sibiricus into a refrigerator, using a prepared phosphate buffer solution (PBS) to dissolve the dry powders, subjecting the solution to filtration sterilization by means of a bacterium filtering membrane, subpackaging the solution and preserving the solution into the refrigerator with the temperature of -20 DEG C for standby. The skin secretions of the tailed amphibian ranodon sibiricus have obvious in-vitro inhibition effects on growth of tumor cells and can be used for preparing antineoplastic drugs.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for purifying the skin secretion of a tailed amphibian distributed in Xinjiang—Xinjiang northern salamander (Ranodon sibiricus) and the application of the secretion in antitumor. Background technique [0002] The skin of amphibians is bare, moist, and vulnerable to the external environment. In the long-term evolution, the skin glands of this kind of organisms have been highly evolved and developed. In addition to keeping the body moist, their functions are crucial to resisting the invasion of microorganisms, viruses, and even natural enemies. Amphibian skin glands are almost all over the body, and can be divided into three types: mucous glands, granular glands, and vesicular glands. The first two types of skin glands account for the vast majority (Lai Ren et al. Amphibian skin active peptides and active biogenic amines[J] . Nature Exploration, 1999, 18(67):71-74). Mucus glan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/56A61P35/00A61K35/65
Inventor 袁亮王秀玲叶小芳周克夫蔡报伟王晗季荣
Owner XINJIANG NORMAL UNIVERSITY
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