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New use of astragalus mongolicus lectin

A Mongolian astragalus and lectin technology, which is applied in the directions of medical preparations containing active ingredients, plant raw materials, peptide/protein components, etc., can solve the problem that there is no antitumor activity of astragalus lectin.

Inactive Publication Date: 2010-02-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no relevant research report on the antitumor activity of astragalus lectin

Method used

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  • New use of astragalus mongolicus lectin
  • New use of astragalus mongolicus lectin
  • New use of astragalus mongolicus lectin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the preparation of Astragalus mongolica lectin

[0025] 1. Preparation of crude extract of Astragalus mongolica

[0026] After crushing 10g of Astragalus mongolica root into 40 mesh powder, add 60mL of 50mmol / L, pH 7.2 phosphate buffer (make the ratio of Astragalus mongolica powder to phosphate buffer 1g: 6mL), and extract with stirring at 4°C 4h. Filter the extract with 4 layers of gauze, centrifuge the filtrate at 10,000×g for 20 min at 4°C, and take the supernatant to obtain the crude extract of Astragalus mongolica.

[0027] 2. Salting out and dialysis of the crude extract

[0028] Add ammonium sulfate powder to the crude extract of Astragalus mongolica obtained in the above step 1 until the saturation is 40%-60%, after stirring for 30 minutes, centrifuge at 12000×g for 15 minutes; take the precipitate, and use 20mmol / L, pH 7.4 Tris-HCl The buffer solution dissolves the precipitate, and then dialyzes overnight in the following solution: the solvent ...

Embodiment 2

[0039] Embodiment 2, preparation of Astragalus mongolica lectin

[0040] 1. Preparation of crude extract of Astragalus mongolica

[0041] The specific preparation method is the same as step 1 of Example 1.

[0042] 2. Salting out and dialysis of the crude extract

[0043] The specific preparation method is the same as step 2 of Example 1.

[0044] 3. ConA-Sepharose 4B affinity chromatography

[0045] First use solution A with pH of 7.4 (the solvent of solution A is the Tris-HCl buffer solution of 20mmol / L, and the solute is that the final concentration is 1mmol / L CaCl 2 , 1mmol / L MnCl 2 , 1mmol / L MgCl 2 and 1mol / L NaCl) pre-equilibrated ConA-Sepharose 4B affinity column, then the supernatant obtained in the above step 2 was loaded on the pre-equilibrated ConA-Sepharose 4B affinity column (1.0 × 3cm), and the The sample flow rate was 0.1 mL / min. After 30min, wash off the unbound protein with the above-mentioned solution A with a pH of 7.4, the flow rate of the buffer is ...

Embodiment 3

[0053] Example 3. Determination of Inhibition of Mongolian Astragali Lectin on Tumor Cell Proliferation in Vitro by MTT Method

[0054] Inoculate HeLa cells, K562 cells, and MG63 cells into 50 mL culture flasks containing 5 mL of culture medium (DMEM culture medium for HeLa cells and MG63, RPMI1640 culture medium for K562 cells), and place at 37 °C, 5% CO 2 cultured in a biochemical incubator; when the cells entered the logarithmic growth phase, they were made into a concentration of about 1×10 5 cells / mL cell suspension, add 200 μL per well into a 96-well plate, place at 37 °C, 5% CO 2 cultured in a biochemical incubator for 24 h.

[0055] The Mongolian astragalus agglutinin obtained in the above-mentioned Example 1 was added to the 96-well plate cultured with HeLa cells, K562 cells and MG63 cells, so that the final concentrations of the mongolian astragalus agglutinin were 0, 2.5, 5, 10, 20 , 40, 60, and 80 μg / mL, each concentration was repeated three times; an equal volum...

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Abstract

The invention discloses new application of astragalus mongolicus lectin, which refers to application of the astragalus mongolicus lectin in inhibiting the growth of tumors, in particular in inhibitingthe proliferations of cervical cancer cells HeLa, leukemia cells K562 and osteosarcoma cells MG63. The astragalus mongolicus lectin is prepared by a method which comprises the following steps: 1) grinding astragalus mongolicus roots and then leaching the grinded astragalus mongolicus roots by the prior method, for instance, leaching the grinded astragalus mongolicus roots with a phosphate buffersolution, and then separating the supernatant to obtain an astragalus mongolicus crude extract; 2) salting the crude extract obtained in the step 1) out, for instance, performing sedimentation on thecrude extract with ammonium sulfate, dissolving sediments with a Tris-HCl buffer solution, performing dialysis on the dissolved solution, and centrifugating the solution obtained after the dialysis toobtain the supernatant; and 3) performing chromatography on the supernatant obtained in the step 2) by a ConA-Sepharose 4B affinity chromatographic column, performing dialysis on the solution obtained after the chromatography and condensing the solution obtained after the dialysis to obtain the astragalus mongolicus lectin.

Description

technical field [0001] The invention relates to a new application of mongolian astragalus lectin. Background technique [0002] Chinese herbal medicines are mainly used according to their active ingredients. In order to make full use of resources and explore new active ingredients, in recent years, many substances with biological activity have been found in various Chinese herbal medicines. Separation and purification of active protein components in Chinese herbal medicines, in order to find more protein sources with biological activity, has become an important research direction of many scholars at home and abroad. Studies by Huang Yiling and others have proved that trichosanthin can inhibit the proliferation and induce apoptosis of human cervical cancer HeLa cells (Huang Yiling et al. Effects of trichosanthin on proliferation and apoptosis of human cervical cancer HeLa cells. Chinese Pharmacology Bulletin. 2005, 21( 2): 253-254). [0003] Astragalus is leguminasae (Legum...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61P35/00A61P35/02A61K36/481
Inventor 闫巧娟江正强李艳霞朱利芬
Owner CHINA AGRI UNIV
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