Fenugreek protein, its separation and extraction method and its application in the preparation of bone marrow hsc clearance auxiliary agent

A technology of fenugreek and adjuvant, applied in the field of natural functional products, can solve the problem of unclear fenugreek plant protein macromolecular function, etc., and achieve the effects of easy industrialization, popularization and application, simple ingredients and rich protein content.

Active Publication Date: 2021-11-30
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the patent does not clarify the function of the fenugreek plant protein macromolecule

Method used

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  • Fenugreek protein, its separation and extraction method and its application in the preparation of bone marrow hsc clearance auxiliary agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The separation and extraction method of fenugreek protein comprises the following steps:

[0031] (1) Grind fenugreek, first soak it in petroleum ether to degrease, then add KH 2 PO 4 -NaOH buffer solution (pH value is 7; the same below) is stirred and extracted for 4-12 hours, centrifuged, and the supernatant is taken; the above-mentioned extraction operation is repeated several times, and the obtained supernatant is combined to be the extract;

[0032] (2) Slowly add ammonium sulfate to 40% saturation in the extract, centrifuge (2000r / min, centrifugal 15min; the same below), collect the supernatant; continue to add ammonium sulfate to 50% saturation, centrifuge, collect precipitation;

[0033] (3) Transfer the precipitate to a dialysis bag, dialyze in deionized water (change the water every 12 hours on average), until the dialysate is colorless, and finally freeze-dry the components in the dialysis bag to obtain fenugreek protein;

[0034] Give the fenugreek protei...

Embodiment 2

[0042] The separation and extraction method of fenugreek protein is the same as in Example 1, and the difference is step (2):

[0043] (2) slowly add ammonium sulfate to 50% saturation to the extract, centrifuge to collect supernatant; continue to add ammonium sulfate to 60% saturation, centrifuge to collect precipitation;

[0044] The fenugreek protein precipitated by ammonium sulfate saturation 50%-60% prepared in this example was given at a dose of 3.0 g / kg.d for 28 days. The effect of myeloablative action.

[0045] The experimental method is the same as in Example 1.

[0046] As can be seen from Table 1, compared with the Bu / Cy group, administration of the fenugreek protein of this example at a dose of 3.0 g / kg.d for 28 days increased the effect on G in the LSK cell population. 0 scavenging, manifested as G in the LSK cell population 0 The proportion of cells decreased significantly. It shows that it has the effect of increasing the myeloablative effect of pretreatment o...

Embodiment 3

[0048] The separation and extraction method of fenugreek protein is with embodiment 1, and difference is step (2):

[0049] (2) slowly adding ammonium sulfate to the extract to a saturation of 60%, centrifuging, and collecting the supernatant; continue adding ammonium sulfate to a saturation of 70%, centrifuging, and collecting the precipitate;

[0050] The fenugreek protein with ammonium sulfate saturation of 60%-70% precipitated prepared in this example was given at a dose of 2.0 g / kg.d for 28 days. The effect of myeloablative action.

[0051] The experimental method is the same as in Example 1.

[0052] As can be seen from Table 1, compared with the Bu / Cy group, administration of the fenugreek protein of this example at a dose of 2.0 g / kg.d for 28 days increased the effect on G in the LSK cell population. 0 scavenging, manifested as G in the LSK cell population 0 The proportion of cells decreased significantly. It shows that it has the effect of increasing the myeloabla...

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Abstract

The invention discloses a fenugreek protein, a method for separating and extracting the fenugreek protein, and an application in preparing a bone marrow HSC clearing auxiliary agent. The method for separating and extracting fenugreek protein comprises the following steps: slowly adding ammonium sulfate to the fenugreek extract to a saturation of 40%, centrifuging, and collecting the supernatant; continuing to add ammonium sulfate to a saturation of 70%, Centrifuge to collect the precipitate; dialyze and freeze-dry the precipitate to obtain fenugreek protein. Fenugreek protein of the present invention can significantly reduce G in the LSK cell population 0 The ratio of cells, which have the function of enhancing bone marrow HSC depletion in bone marrow transplantation preconditioning of thalassemia iron overloaded mice, can be used as an adjuvant for bone marrow HSC depletion in thalassemia bone marrow transplantation preconditioning.

Description

technical field [0001] The invention belongs to the field of natural functional products, and in particular relates to fenugreek protein, its separation and extraction method and its application in the preparation of bone marrow HSC (hematopoietic stem cell) clearance auxiliary agent. Background technique [0002] Thalassemia is one of the common hereditary blood diseases. Its molecular basis is globin gene defect leading to the disorder of globin chain synthesis, which makes the ratio of α-chain / non-α-chain forming hemoglobin tetramer unbalanced, which in turn causes red blood cell destruction. Hemolysis. Routine red blood cell transfusion and iron removal therapy. For patients with β-thalassemia major, bone marrow transplantation is considered to be the only curative method at present. [0003] Preconditioning is one of the important steps in bone marrow transplantation. Its main purpose is to: ① eradicate abnormal bone marrow hematopoietic stem cells (HSCs) and minimize...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14C07K1/30A61K38/02A61P37/06A61P7/06A23L33/185A23L2/66A21D2/26A21D13/064
CPCA21D2/266A21D13/064A23L2/66A23V2002/00A61K38/00A23L33/185A61P7/06A61P37/06C07K1/14C07K1/145C07K1/30A23V2200/30A23V2250/548
Inventor 李思瑶刘备王芳李续娥
Owner SOUTH CHINA NORMAL UNIVERSITY
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