New application of astragalus polysaccharide in chromosome protection

A technology of astragalus polysaccharides and chromosomes, applied in the field of biomedicine, can solve the problems of unreported formaldehyde toxicity protection effect and the like

Inactive Publication Date: 2019-12-24
GANSU UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, domestic research on the anti-tumor effect of its main active ingredient, astragalus polysac...

Method used

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  • New application of astragalus polysaccharide in chromosome protection
  • New application of astragalus polysaccharide in chromosome protection
  • New application of astragalus polysaccharide in chromosome protection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: cell culture and grouping

[0022] 1) Preparation of formaldehyde exposure solution: According to the content (M / V) of formaldehyde is 37%, relative molecular weight is 30.03, so 0.1194mL of 37% formaldehyde is added to 99.8804mL of BM-MSCs special medium to obtain 100mL Formaldehyde with a concentration of 16mmol / L was sterilized by suction filtration with a 0.22μm filter membrane, aliquoted, and stored at 4°C. Dilute to 120μmol / L working solution concentration when used.

[0023] 2) Preparation of astragalus polysaccharide solution: take 20 mg of astragalus polysaccharide powder (purity ≥ 98%), dissolve it in 10 mL of DMEM / F12 medium, filter through a 0.22 μm microporous filter to make the final concentration of astragalus polysaccharide to 2 mg / mL, and store at 4°C , diluted with time.

[0024] Human BM-MSCs use complete medium for human BM-MSCs, placed in 5% CO 2 , 37°C, and a saturated humidity incubator. The medium was changed every 2-3 days, and...

Embodiment 2

[0025] Embodiment 2: Observation of cell morphology

[0026] Observe human BM-MSCs under a phase-contrast inverted microscope, take pictures with a microscopic imaging system, and record the results.

[0027] Such as figure 1 As shown, the BM-MSCs in the blank group grew adherently, showing a long fusiform or star shape, relatively uniform in size, and arranged tightly in a spiral shape. In the formaldehyde group, the number of cells decreased, some cells were suspended, the volume was reduced and the size was different, and they were round or even irregular. APS 0.4, 1.0, 4.0 mg·mL -1 In the formaldehyde group, the shape of the cells was significantly restored. Compared with the formaldehyde group, the number of cells increased significantly, and a small number of cells were suspended and round. -1 The APS group was the most obvious.

Embodiment 3

[0028] Embodiment 3: micronucleus test detects the formation of micronuclei

[0029] Trypsinize, collect cells, hypotonic with KCl solution for 3min, fix, drop slices, and Giemsa staining. 1000 cells were randomly observed on each slide, most of the micronuclei contained in the cells were round, with smooth and tidy edges, 1 / 16-1 / 3 of the diameter of the main nucleus, non-refractive, not connected with the main nucleus, and chromophilic The sex is consistent with the nucleoplasm, which is purple or blue-purple, and one or more micronuclei can appear in a cell. The number of cells with micronuclei was counted, and the rate of micronucleated cells (‰)=the total number of cells with micronuclei / total number of observed cells×1000‰.

[0030] The results of micronucleus experiments showed that ( figure 2, Table 1), the micronucleus rate of BM-MSCs formed by formaldehyde was significantly higher than that of the blank group (P-1 After the action, the rate of micronuclei was signi...

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Abstract

The invention discloses a protection effect and a potential mechanism of astragalus polysaccharide (APS) on micronucleus formation of bone-marrow mesenchymal stem cells (BM-MSCs) of formaldehyde-contaminated human, and frequency increase of sister chromatid exchanges (SCE), provides a new application of the astragalus polysaccharide in chromosome protection, and provides a basis for the astragaluspolysaccharide to protect the toxic effect of formaldehyde on organisms. The invention determines that the astragalus polysaccharide can maintain morphological characteristics and the cell number ofhuman BM-MSCs in a formaldehyde environment. The invention determines that the astragalus polysaccharide can reduce micronucleus formation of human BM-MSCs in a formaldehyde environment. The inventiondetermines that the astragalus polysaccharide can reduce the SCE frequency of human BM-MSCs in a formaldehyde environment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a new application of astragalus polysaccharide chromosome protection. Background technique [0002] Human bone marrow mesenchymal stem cells (bone marrow mesenchymal stem cells, BM-MSCs) BM-MSCs are stem cells that exist in the bone marrow and have multi-lineage differentiation potential. stability is of great significance. Formaldehyde, as an environmental pollutant, can cause toxic damage to cells. It has been reported that formaldehyde can induce increased chromosomal damage in human bone marrow blood progenitor cells, and can also induce the formation of micronuclei and the formation of micronuclei in polychromatic erythrocytes and BM-MSCs in mouse bone marrow. The increased frequency of sister chromatid exchanges (sister chromatidexchanges, SCE) provides a biological basis for formaldehyde to cause leukemia. [0003] Micronucleus is the acentric fragment of...

Claims

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Application Information

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IPC IPC(8): A61K31/715A61P39/02
CPCA61K31/715A61P39/02
Inventor 舍雅莉李亚玲刘永琦李长天
Owner GANSU UNIV OF CHINESE MEDICINE
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