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Sea cucumber polypeptide, preparation method and application thereof

The technology of sea cucumber polypeptide and sea cucumber is applied in the field of sea cucumber polypeptide and its preparation, and can solve the problems such as the treatment of bone marrow cell proliferation effect not seen by sea cucumber polypeptide.

Inactive Publication Date: 2009-08-26
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the effect of peptides isolated and purified from sea cucumbers on increasing the number of white blood cells and promoting the proliferation of bone marrow cells after chemotherapy. Therapeutic reports of proliferative effects

Method used

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  • Sea cucumber polypeptide, preparation method and application thereof
  • Sea cucumber polypeptide, preparation method and application thereof
  • Sea cucumber polypeptide, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Extracting sea cucumber polypeptides from sea cucumbers, the steps are as follows:

[0023] (1) Take fresh sea cucumber, remove internal organs, wash, and break into 1cm 3 For the left and right small pieces, add 8-10 times of water to homogenate.

[0024] The homogenate was subjected to pepsin (the amount of enzyme was 0.4% of the wet weight of sea cucumber, pH 2.0-2.5, temperature 50°C, time 5h), trypsin (the amount of enzyme was 0.45% of the wet weight of sea cucumber, pH 8.0-8.5, Temperature 50°C, time 5h) enzymolysis, enzymolysis is completed, boiling inactivation, centrifugation (10000r / min, 10min) to take the supernatant.

[0025] The supernatant was precipitated with 60% ethanol overnight, centrifuged (4000r / min, 15min) to take the supernatant, concentrated under reduced pressure and freeze-dried to obtain the crude sea cucumber polypeptide, which was named HST.

[0026] (2) Dissolve the crude sea cucumber polypeptide in distilled water, perform Sep...

Embodiment 2

[0028] Example 2: The effect of crude sea cucumber polypeptide on increasing the number of white blood cells in immunodeficiency mice after chemotherapy

[0029] Fifty Kunming mice (body weight 20±1.0 g) were selected and randomly divided into 5 groups, 10 mice in each group, half male and half male, and cultured separately. The blank control group, the model control group, and the HST low, medium, and high dose groups were followed in turn.

[0030] Except for the blank control group, the other 4 groups were all modeled with cyclophosphamide, and each mouse was intraperitoneally injected 4 times at 30 mg / kg.bw.

[0031] From the next day after modeling, each group was given intragastric administration once a day, with a volume of 0.2ml / 10g.bw, for 10 consecutive days. The blank control group and the model control group used normal saline; the sea cucumber peptide group used the corresponding sea cucumber peptide components (according to the conversion relationship between mo...

Embodiment 3

[0037] Example 3: The effect of HST-2 on increasing the number of white blood cells in immunodeficient mice after chemotherapy

[0038] 110 Kunming mice (body weight 20±1.0 g) were selected and randomly divided into 11 groups, 10 mice in each group, with half male and half male, and were cultured separately. The blank control group, model control group, HST-1 low, medium and high dose groups, HST-2 low, medium and high dose groups, HST-3 low, medium and high dose groups.

[0039] Except for the blank control group, the other 10 groups were all modeled with cyclophosphamide, and each mouse was intraperitoneally injected 4 times at 30 mg / kg.bw.

[0040] From the next day after modeling, each group was given intragastric administration once a day, with a volume of 0.2ml / 10g.bw, for 10 consecutive days. The blank control group and the model control group used normal saline; the sea cucumber peptide group used the corresponding sea cucumber peptide components (according to the con...

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Abstract

The invention discloses a sea cucumber polypeptide, wherein a preparation method thereof comprises the following steps: (1) getting fresh stichopus japonicus, removing entrails, cleaning and crushing the stichopus japonicus into small blocks, and adding water to form homogenate; after the enzymolysis of the homogenate liquid, centrifugating and getting the supernatant; adding ethanol, standing, centrifugating and getting the supernatant, concentrating the supernatant at reducing pressure and drying the concentrate to form a crude product of the sea cucumber polypeptide; (2) dissolving the crude product of the sea cucumber polypeptide by using distilled water, carrying out gel filtration chromatography by Sephadex LH-20, eluting the sea cucumber polypeptide by double-distilled water, getting a second peak, collecting active components, and condensing, freezing and freeze-drying the active components; and (3) dissolving the active components by double-distilled water, carrying out ion exchange chromatography by Q Sepharose Fast Flow, linearly eluting the active components by NaCl solution, desalting, getting the second peak, collecting the active components, and condensing, freeze-drying the active components to form the sea cucumber polypeptide. The sea cucumber polypeptide can be used for preparing medicaments or health-care products for increasing leukocytes and also can be used for preparing medicaments or health-care products for multiplying marrow cells after chemotherapy.

Description

technical field [0001] The invention relates to a sea cucumber polypeptide and its preparation method and application. Background technique [0002] With the increasingly serious environmental and food pollution in modern society, the incidence of cancer is getting higher and higher, and the main methods of treating cancer are radiotherapy and chemotherapy, but their side effects are very large, and most of them will cause leukopenia. [0003] There have been many reports on the important medicinal value of sea cucumber and its biologically active substances. The currently reported active substances are mainly polysaccharides, polypeptides and sea cucumber saponins, etc.; the effects of these active substances mainly include anticoagulant and hemostatic effects, anti-tumor effects, enhancing immunity, delaying aging, lowering blood sugar, anti-radiation, and anti-inflammation. However, there is no report on the effect of peptides isolated and purified from sea cucumbers on ...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/435A61K38/17A23L1/305A61P35/00A61P43/00A23L33/18
Inventor 吉爱国梁浩王允山宋淑亮逄少堃
Owner SHANDONG UNIV
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